@article{liu_paudel_flowers_piedrahita_wang_2023, title={Adrenomedullin Stimulates Proliferation, Migration and Adhesion of Porcine Trophectoderm Cells Via CALCRL-AKT-TSC2-MTORC1 Cell Signaling Pathway}, volume={101}, ISSN={["1525-3163"]}, DOI={10.1093/jas/skad068.040}, abstractNote={
 Adrenomedullin (ADM) as a highly conserved peptide hormone has been reported to increase significantly in the uterine lumen during the peri-implantation period of pregnancy in pigs, but its functional roles in growth and development of porcine conceptus (embryonic/fetus and its extra-embryonic membranes) as well as underlying mechanisms remain largely unknown. Therefore, we conducted in vitro experiments using our established porcine trophectoderm cell line (pTr1) isolated from day -12 porcine conceptuses to test the hypothesis that porcine ADM stimulates cell proliferation, migration and adhesion via AKT-TSC2-MTOR cell signaling pathway in pTr1 cells. The pTr1 cells were cultured in DMEM/F12 medium with 10% fetal bovine serum (FBS), 50 U/mL penicillin, 50 μg/mL streptomycin, 0.1 mM each for nutritionally nonessential amino acids, 1 mM sodium pyruvate, 2 mM glutamine, and 4 μg/mL insulin. Opti-MEM supplied with 2.5% (vol/vol) charcoal-stripped FBS was used for siRNA-mediated knockdown targeting non-treated control (siNTC) or specific genes including ADM (siADM) and its shared receptor component calcitonin-receptor-like receptor (CALCRL; siCALCRL). Cells were starved in FBS- and insulin-free medium for 24 hours before treatment. For proliferation assay, cell numbers were determined by staining with Janus-Green B after 48 h incubation. For migration assay, cells were treated with ADM after straight scratch in 6-well plates, and area of cell migration was calculated after 12 h treatment. For adhesion assay, cells were trypsinized in T-25 flasks and allowed for seeding in 96-well plates with density of 2×105 cells/0.2 mL/well, and the numbers of attached cells were determined after 12 h incubation. Western blot analyses were used to determine the expressions of target proteins at total and phosphorylated level. Porcine ADM at 10-7 M stimulated (P < 0.05) pTr1 cell proliferation, migration and adhesion by 1.4%-, 1.5%- and 1.2%-folds, respectively. These ADM-induced effects were abrogated (P < 0.05) by siADM and siCALCRL, as well as by rapamycin, the inhibitor of mechanistic target of rapamycin (MTOR). Using siRNA-mediated knockdown of CALCRL coupled with Western blot analyses, ADM signaling transduction was determined in which ADM binds to CALCRL to increase phosphorylation of MTOR, its downstream effectors (4EBP1, P70S6K, and S6), and upstream regulators (AKT and TSC2). Collectively, these results suggest that porcine ADM in histotroph act on its receptor component CALCRL to activate AKT-TSC2-MTOR, particularly MTORC1 signaling cascade, leading to elongation, migration and attachment of conceptuses. This research was supported by Agriculture and Food Research Initiative Competitive Grant no. 2022-67015-36491 from the USDA National Institute of Food and Agriculture.}, journal={JOURNAL OF ANIMAL SCIENCE}, author={Liu, Bangmin and Paudel, Sudikshya and Flowers, William L. and Piedrahita, Jorge A. and Wang, Xiaoqiu}, year={2023}, month={May} }
 @article{paudel_liu_cummings_wang_2023, title={Adrenomedullin: A Novel Peptide Hormone for Uterine Receptivity and Conceptus Elongation in Pigs?}, volume={101}, ISSN={["1525-3163"]}, DOI={10.1093/jas/skad068.049}, abstractNote={
 Adrenomedullin (ADM) is an evolutionarily conserved multi-functional peptide hormone in mammalian species. The implantation sites of conceptuses (embryo/fetus and its extra-embryonic membranes) in pregnant Adm+/− female mice are abnormally spaced leading to fetal crowding, intrauterine growth restriction (IUGR) and increases in pregnancy loss. As litter-bearing species, pigs exhibit a high incidence of early embryonic death (30-40%) and naturally occurring IUGR (15-25%), and significant numbers of stillborn piglets (3-9%). Therefore, it is imperative that the roles of ADM in regulation of uterine receptivity, as well as growth and development of conceptus in pigs be established. This study determined abundances of ADM in uterine luminal fluid, and the patterns of expression of ADM and its receptor components in uteri from cyclic and pregnant gilts, as well as conceptuses during the peri-implantation period of pregnancy. ADM receptor components include: calcitonin receptor-like receptor (CALCRL; G protein-coupled receptor bound by ADM), receptor activity modifying protein (RAMP2; dimerized with CALCRL to form ADM1 receptor) and RAMP3 (dimerized with CALCRL to form ADM2 receptor with lower specificity to ADM) and atypical chemokine receptor 3 (ACKR3; a decoy receptor that serves as a cell-autonomous molecular rheostat to dampen ADM signaling). Total recoverable ADM was greater in the uterine fluid of pregnant compared with cyclic gilts between days 10 and 16 post-estrus and was from uterine luminal epithelial (LE) and conceptus trophectoderm (Tr) cells. Uterine expression of CALCRL, RAMP2, and ACKR3 were affected by day (P < 0.05), pregnant status (P < 0.01) and/or day x status (P < 0.05). Within porcine conceptuses, expression of CALCRL, RAMP2 and ACKR3 increased between days 10 and 16 of pregnancy. Using an established porcine trophectoderm (pTr1) cell line isolated from elongating porcine conceptuses recovered on day 2 post-breeding, it was determined that 10-7 M ADM stimulated proliferation of pTr1 cells (P < 0.05) at 48 h, and increased phosphorylated mechanistic target of rapamycin (p-MTOR) and 4E binding protein (p-4EBP1) by 6.1- and 4.9-fold (P < 0.0001), respectively. These novel results indicate a significant role for ADM in uterine receptivity for implantation and conceptus growth and development in pigs. They also provide a framework for future studies of ADM signaling to affect proliferation and migration of Tr cells, spacing of blastocysts, implantation and placentation in pigs. This research was supported by Agriculture and Food Research Initiative Competitive Grant no. 2022-67015-36491 from the USDA National Institute of Food and Agriculture.}, journal={JOURNAL OF ANIMAL SCIENCE}, author={Paudel, Sudikshya and Liu, Bangmin and Cummings, Magdalina J. and Wang, Xiaoqiu}, year={2023}, month={May} }
 @article{liu_duan_liu_wang_2023, title={Metabolomic Analysis of Uterine Luminal Fluid During the Peri-Implantation Period of Pregnancy in Pigs}, volume={101}, ISSN={["1525-3163"]}, DOI={10.1093/jas/skad068.046}, abstractNote={
 During the peri-implantation period of pregnancy in pigs, the rapid elongation of conceptuses (embryonic/fetus and its extraembryonic membranes) is highly dependent on the composition of histotroph secreted from uterine luminal (LE) and glandular (GE) epithelial and stroma cells, as well as selective transport of nutrients. However, little is known about the metabolites in the uterine luminal fluid (ULF) associated with the uterine-conceptus communication during early pregnancy in pigs. Thus, this study was conducted to profile the metabolome in porcine ULF between days 10 and 16 of estrus cycle (C) and pregnancy (P). Gilts were observed for estrus and/or bred via artificial insemination at 12 and 24 h after onset of estrus (day 0). On days 10, 12, 14, or 16 of the estrous cycle and pregnancy (n = 6 gilts per day and status), uteri were flushed with 20 mL sterile PBS (pH 7.2) after gilts were subjected to a midventral laparotomy. Pregnancy was confirmed by the presence of one or more morphologically normal conceptuses. Recoverable uterine flushings (i.e., ULF) were then subjected to metabolomic analysis by LC-MS. Overall, 222 metabolites were detected in the ULF of which 102 were altered by status and 63 were altered by days (P < 0.05; fold change>1.5). Comparing to cyclic day 10 (10C), pregnancy stimulated increases in citric acid, lysine, arginine and other 19 metabolites in the ULF at pregnant day 10 (10P). At 12P when porcine conceptus initiates its morphological changes, ornithine, α-D-glucose, phenylalanine and other 13 metabolites were up-regulated as compared with 12C. At 14P when conceptus initiates implantation, 15 metabolites changed in ULF. At 16P, 123 metabolites were altered in ULF as compared with 16C. Interestingly, of 15 altered metabolites in ULF, 12 were downregulated including asparagine, nicotinamide riboside, and citrulline at 14P. Whereas 102 of 123 altered metabolites were upregulated including phosphorylcholine, 6-phosphate fructose and 6-phosphate glucose in ULF at 16P. To determine the absolute amount of amino acids in the ULF, we performed the targeted metabolomic analyses for 19 amino acids. As a result, arginine, asparagine, glutamic acid, glutamine, histidine, leucine/isoleucine, lysine, phenylalanine, proline, and valine were increased between 10P and 16P. They were less expressed in the ULF of estrus cycle as compared with pregnancy, and remained unchanged between 10C and 16C. Pathway analysis based on KEGG database indicating that arginine biosynthesis, alanine, aspartate and glutamate metabolism, and aminoacyl-tRNA biosynthesis have the highest degree of enrichment. These novel findings provide the foundation for future investigation of metabolomic and functional studies on both endometrium and conceptus required for elongation of porcine conceptus during peri-implantation period of pregnancy. This research was supported by Agriculture and Food Research Initiative Competitive Grant no. 2022-67015-36491 from the USDA National Institute of Food and Agriculture}, journal={JOURNAL OF ANIMAL SCIENCE}, author={Liu, Bangmin and Duan, Likun and Liu, Xiaojing and Wang, Xiaoqiu}, year={2023}, month={May} }
 @article{wang_man_wang_odle_maltecca_lin_2023, title={MicroRNA and mRNA sequencing analyses reveal key hepatic metabolic and signaling pathways responsive to maternal undernutrition in full-term fetal pigs}, volume={116}, ISSN={["1873-4847"]}, url={https://doi.org/10.1016/j.jnutbio.2023.109312}, DOI={10.1016/j.jnutbio.2023.109312}, abstractNote={Maternal undernutrition is highly prevalent in developing countries, leading to severe fetus/infant mortality, intrauterine growth restriction, stunting, and severe wasting. However, the potential impairments of maternal undernutrition to metabolic pathways in offspring are not defined completely. In this study, 2 groups of pregnant domestic pigs received nutritionally balanced gestation diets with or without 50% feed intake restriction from 0 to 35 gestation days and 70% from 35 to 114 gestation days. Full-term fetuses were collected via C-section on day 113/114 of gestation. MicroRNA and mRNA deep sequencing were analyzed using the Illumina GAIIx system on fetal liver samples. The mRNA-miRNA correlation and associated signaling pathways were analyzed via CLC Genomics Workbench and Ingenuity Pathway Analysis Software. A total of 1189 and 34 differentially expressed mRNA and miRNAs were identified between full-nutrition (F) and restricted-nutrition (R) groups. The correlation analyses showed that metabolic and signaling pathways such as oxidative phosphorylation, death receptor signaling, neuroinflammation signaling pathway, and estrogen receptor signaling pathways were significantly modified, and the gene modifications in these pathways were associated with the miRNA changes induced by the maternal undernutrition. For example, the upregulated (P<.05) oxidative phosphorylation pathway in R group was validated using RT-qPCR, and the correlational analysis indicated that miR-221, 103, 107, 184, and 4497 correlate with their target genes NDUFA1, NDUFA11, NDUFB10 and NDUFS7 in this pathway. These results provide the framework for further understanding maternal malnutrition's negative impacts on hepatic metabolic pathways via miRNA-mRNA interactions in full-term fetal pigs.}, journal={JOURNAL OF NUTRITIONAL BIOCHEMISTRY}, author={Wang, Feng and Man, Chaolai and Wang, Xiaoqiu and Odle, Jack and Maltecca, Christian and Lin, Xi}, year={2023}, month={Jun} }
 @article{liu_paudel_flowers_piedrahita_wang_2023, title={Uterine histotroph and conceptus development: III. Adrenomedullin stimulates proliferation, migration and adhesion of porcine trophectoderm cells via AKT-TSC2-MTOR cell signaling pathway}, volume={4}, ISSN={["1438-2199"]}, DOI={10.1007/s00726-023-03265-6}, abstractNote={Adrenomedullin (ADM) as a highly conserved peptide hormone has been reported to increase significantly in the uterine lumen during the peri-implantation period of pregnancy in pigs, but its functional roles in growth and development of porcine conceptus (embryonic/fetus and its extra-embryonic membranes) as well as underlying mechanisms remain largely unknown. Therefore, we conducted in vitro experiments using our established porcine trophectoderm cell line (pTr2) isolated from Day-12 porcine conceptuses to test the hypothesis that porcine ADM stimulates cell proliferation, migration and adhesion via activation of mechanistic target of rapamycin (MTOR) cell signaling pathway in pTr2 cells. Porcine ADM at 10–7 M stimulated (P < 0.05) pTr2 cell proliferation, migration and adhesion by 1.4-, 1.5- and 1.2-folds, respectively. These ADM-induced effects were abrogated (P < 0.05) by siRNA-mediated knockdown of ADM (siADM) and its shared receptor component calcitonin-receptor-like receptor (CALCRL; siCALCRL), as well as by rapamycin, the inhibitor of MTOR. Using siRNA-mediated knockdown of CALCRL coupled with Western blot analyses, ADM signaling transduction was determined in which ADM binds to CALCRL to increase phosphorylation of MTOR, its downstream effectors (4EBP1, P70S6K, and S6), and upstream regulators (AKT and TSC2). Collectively, these results suggest that porcine ADM in histotroph acts on its receptor component CALCRL to activate AKT-TSC2-MTOR, particularly MTORC1 signaling cascade, leading to elongation, migration and attachment of conceptuses.}, journal={AMINO ACIDS}, author={Liu, Bangmin and Paudel, Sudikshya and Flowers, William L. and Piedrahita, Jorge A. and Wang, Xiaoqiu}, year={2023}, month={Apr} }
 @article{duan_scheidemantle_lodge_cummings_pham_wang_kennedy_liu_2022, title={Prioritize biologically relevant ions for data-independent acquisition (BRI-DIA) in LC-MS/MS-based lipidomics analysis}, volume={18}, ISSN={["1573-3890"]}, DOI={10.1007/s11306-022-01913-8}, abstractNote={Data-dependent acquisition (DDA) is the most commonly used MS/MS scan method for lipidomics analysis on orbitrap-based instrument. However, MS instrument associated software decide the top N precursors for fragmentation, resulting in stochasticity of precursor selection and compromised consistency and reproducibility. We introduce a novel workflow using biologically relevant lipids to construct inclusion list for data-independent acquisition (DIA), named as BRI-DIA workflow.To ensure consistent coverage of biologically relevant lipids in LC-MS/MS-based lipidomics analysis.Biologically relevant ion list was constructed based on LIPID MAPS and lipidome atlas in MS-DIAL 4. Lipids were extracted from mouse tissues and used to assess different MS/MS scan workflow (DDA, BRI-DIA, and hybrid mode) on LC-Orbitrap Exploris 480 mass spectrometer.DDA resulted in more MS/MS events, but the total number of unique lipids identified by three methods (DDA, BRI-DIA, and hybrid MS/MS scan mode) is comparable (580 unique lipids across 44 lipid subclasses in mouse liver). Major cardiolipin molecular species were identified by data generated using BRI-DIA and hybrid methods and allowed calculation of cardiolipin compositions, while identification of the most abundant cardiolipin CL72:8 was missing in data generated using DDA method, leading to wrong calculation of cardiolipin composition.The method of using inclusion list comprised of biologically relevant lipids in DIA MS/MS scan is as efficient as traditional DDA method in profiling lipids, but offers better consistency of lipid identification, compared to DDA method. This study was performed using Orbitrap Exploris 480, and we will further evaluate this workflow on other platforms, and if verified by future work, this biologically relevant ion fragmentation workflow could be routinely used in many studies to improve MS/MS identification capacities.}, number={8}, journal={METABOLOMICS}, author={Duan, Likun and Scheidemantle, Grace and Lodge, Mareca and Cummings, Magdalina J. and Pham, Eva and Wang, Xiaoqiu and Kennedy, Arion and Liu, Xiaojing}, year={2022}, month={Jul} }
 @article{cummings_yu_paudel_hu_li_hemberger_wang_2022, title={Uterine-specific SIRT1 deficiency confers premature uterine aging and impairs invasion and spacing of blastocyst, and stromal cell decidualization, in mice}, volume={28}, ISSN={["1460-2407"]}, DOI={10.1093/molehr/gaac016}, abstractNote={A distinct age-related alteration in the uterine environment has recently been identified as a prevalent cause of the reproductive decline in older female mice. However, the molecular mechanisms that underlie age-associated uterine adaptability to pregnancy are not known. Sirtuin 1 (SIRT1), a multifunctional NAD+-dependent deacetylase that regulates cell viability, senescence and inflammation during aging, is reduced in aged decidua. Thus, we hypothesize that SIRT1 plays a critical role in uterine adaptability to pregnancy and that uterine-specific ablation of Sirt1 gene accelerates premature uterine aging. Female mice with uterine ablation of Sirt1 gene using progesterone receptor Cre (PgrCre) exhibit subfertility and signs of premature uterine aging. These Sirt1-deficient mothers showed decreases in litter size from their 1st pregnancy and became sterile (25.1 ± 2.5 weeks of age) after giving birth to the third litter. We report that uterine-specific Sirt1 deficiency impairs invasion and spacing of blastocysts, and stromal cell decidualization, leading to abnormal placentation. We found that these problems traced back to the very early stages of hormonal priming of the uterus. During the window of receptivity, Sirt1 deficiency compromises uterine epithelial-stromal crosstalk, whereby estrogen, progesterone and Indian hedgehog signaling pathways are dysregulated, hampering stromal cell priming for decidualization. Uterine transcriptomic analyses also link these causes to perturbations of histone proteins and epigenetic modifiers, as well as adrenomedullin signaling, hyaluronic acid metabolism, and cell senescence. Strikingly, our results also identified genes with significant overlaps with the transcriptome of uteri from aged mice and transcriptomes related to master regulators of decidualization (e.g. Foxo1, Wnt4, Sox17, Bmp2, Egfr and Nr2f2). Our results also implicate accelerated deposition of aging-related fibrillar Type I and III collagens in Sirt1-deficient uteri. Collectively, SIRT1 is an important age-related regulator of invasion and spacing of blastocysts, as well as decidualization of stromal cells.}, number={7}, journal={MOLECULAR HUMAN REPRODUCTION}, author={Cummings, Magdalina J. and Yu, Hongyao and Paudel, Sudikshya and Hu, Guang and Li, Xiaoling and Hemberger, Myriam and Wang, Xiaoqiu}, year={2022}, month={Jun} }
 @article{paudel_wu_wang_2021, title={Amino Acids in Cell Signaling: Regulation and Function}, volume={1332}, ISBN={["978-3-030-74179-2"]}, ISSN={["2214-8019"]}, DOI={10.1007/978-3-030-74180-8_2}, abstractNote={Amino acids are the main building blocks for life. Aside from their roles in composing proteins, functional amino acids and their metabolites play regulatory roles in key metabolic cascades, gene expressions, and cell-to-cell communication via a variety of cell signaling pathways. These metabolic networks are necessary for maintenance, growth, reproduction, and immunity in humans and animals. These amino acids include, but are not limited to, arginine, glutamine, glutamate, glycine, leucine, proline, and tryptophan. We will discuss these functional amino acids in cell signaling pathways in mammals with a particular emphasis on mTORC1, AMPK, and MAPK pathways for protein synthesis, nutrient sensing, and anti-inflammatory responses, as well as cell survival, growth, and development.}, journal={AMINO ACIDS IN NUTRITION AND HEALTH: AMINO ACIDS IN GENE EXPRESSION, METABOLIC REGULATION, AND EXERCISING PERFORMANCE}, author={Paudel, Sudikshya and Wu, Guoyao and Wang, Xiaoqiu}, year={2021}, pages={17–33} }
 @article{paudel_liu_cummings_quinn_bazer_caron_wang_2021, title={Temporal and spatial expression of adrenomedullin and its receptors in the porcine uterus and peri-implantation conceptuses}, volume={105}, ISSN={["1529-7268"]}, DOI={10.1093/biolre/ioab110}, abstractNote={Abstract Adrenomedullin (ADM) is an evolutionarily conserved multifunctional peptide hormone that regulates implantation, embryo spacing, and placentation in humans and rodents. However, the potential roles of ADM in implantation and placentation in pigs, as a litter-bearing species, are not known. This study determined abundances of ADM in uterine luminal fluid, and the patterns of expression of ADM and its receptor components (CALCRL, RAMP2, RAMP3, and ACKR3) in uteri from cyclic and pregnant gilts, as well as conceptuses (embryonic/fetus and its extra-embryonic membranes) during the peri-implantation period of pregnancy. Total recoverable ADM was greater in the uterine fluid of pregnant compared with cyclic gilts between Days 10 and 16 post-estrus and was from uterine luminal epithelial (LE) and conceptus trophectoderm (Tr) cells. Uterine expression of CALCRL, RAMP2, and ACKR3 were affected by day (P < 0.05), pregnant status (P < 0.01) and/or day x status (P < 0.05). Within porcine conceptuses, the expression of CALCRL, RAMP2, and ACKR3 increased between Days 10 and 16 of pregnancy. Using an established porcine trophectoderm (pTr1) cell line, it was determined that 10–7 M ADM stimulated proliferation of pTr1 cells (P < 0.05) at 48 h, and increased phosphorylated mechanistic target of rapamycin (p-MTOR) and 4E binding protein 1 (p-4EBP1) by 6.1- and 4.9-fold (P < 0.0001), respectively. These novel results indicate a significant role for ADM in uterine receptivity for implantation and conceptus growth and development in pigs. They also provide a framework for future studies of ADM signaling to affect proliferation and migration of Tr cells, spacing of blastocysts, implantation, and placentation in pigs. Summary sentence Our study suggests that adrenomedullin plays a significant role in uterine receptivity and conceptus growth and development, via the stimulation of cell proliferation and activation of MTORC1 signaling pathway in the porcine conceptus trophectoderm. Graphical Abstract}, number={4}, journal={BIOLOGY OF REPRODUCTION}, author={Paudel, Sudikshya and Liu, Bangmin and Cummings, Magdalina J. and Quinn, Kelsey E. and Bazer, Fuller W. and Caron, Kathleen M. and Wang, Xiaoqiu}, year={2021}, month={Oct}, pages={876–891} }
 @article{moustafa_young_wang_wu_li_wang_spencer_demayo_2019, title={ENDOMETRIAL EPITHELIAL FOXO1 DIRECTLY MODULATES SIGNALING PATHWAYS NECESSARY FOR UTERINE RECEPTIVITY.}, volume={112}, ISSN={["1556-5653"]}, DOI={10.1016/j.fertnstert.2019.07.913}, abstractNote={We have previously shown that endometrial FOXO1 transcription factor protein expression is indispensable for murine embryo implantation. We also demonstrated that FOXO1 protein expression is concentrated in the uterine epithelial nuclei during the window of receptivity1. The objective of this study is to better understand the relevance of FOXO1 in human endometrial receptivity. Differentially expressed genes (DEGs) from proliferative (non-receptive) versus mid-secretory (receptive) human endometrial epithelium were compared with DEGs of uterine-specific FOXO1d/d versus FOXO1f/f mice to determine which FOXO1-regulated murine genes are also regulated during the transition to receptive human endometrium. Ingenuity Pathway Analysis (IPA) identified regulated pathway and FOXO1 ChIP-seq identified genes directly bound by FOXO1. Mouse uterine epithelium was isolated by laser capture on day 4.5 of natural pregnancy. The transcriptome of FOXO1 knockout epithelium was compared to that of wildtype mice using RNA sequencing, identifying DEGs defined by at least a 2-fold change. RNA sequencing was also used to generate DEGs from enzymatically separated human endometrial epithelium between non-receptive and receptive by at least a 1.5-fold change2. IPA of the FOXO1 altered murine genes conserved in human phase-related DEGs was used to identify altered pathways with known roles in endometrial receptivity. Subsequent comparison of the components of each pathway with FOXO1 ChIP-seq identified the direct role of FOXO1 on the altered pathway. 1301 RNA species were common to both the mouse and human DEGs. 318 of these genes were directly bound by FOXO1. Pathway analysis identified Wnt/B-catenin signaling (-log(p-value) 4.14, Z score -0.626), estrogen mediated proliferation (-log(p-value) 10, Z score -3.05), and IL-6 signaling (-log(p-value) 4.73, Z score 0.943) as significantly altered by both human cycle phase and murine FOXO1 deletion, strongly suggesting a role of FOXO1 in these critical pathways for normal endometrial function. ChIP-Seq demonstrated direct FOXO1 binding to multiple regulated genes involved in estrogen-mediated proliferation, IL-6 signaling, and Wnt/b-catenin signaling, supporting a direct action of FOXO1 on these essential signaling pathways. FOXO1 was also found to directly bind several upstream regulators critical to endometrial receptivity, including CEBPB and CCND1. Epithelial FOXO1 directly regulates key pathways necessary for human uterine receptivity.}, number={3}, journal={FERTILITY AND STERILITY}, author={Moustafa, Sarah M. and Young, Steven L. and Wang, Tianyuan and Wu, Steve and Li, Rong and Wang, Xiaoqiu and Spencer, Tom and Demayo, Francesco}, year={2019}, month={Sep}, pages={E314–E315} }
 @article{vasquez_wang_wetendorf_franco_mo_wang_lanz_young_lessey_spencer_et al._2018, title={FOXO1 regulates uterine epithelial integrity and progesterone receptor expression critical for embryo implantation}, volume={14}, ISSN={["1553-7404"]}, DOI={10.1371/journal.pgen.1007787}, abstractNote={Successful embryo implantation requires a receptive endometrium. Poor uterine receptivity can account for implantation failure in women who experience recurrent pregnancy loss or multiple rounds of unsuccessful in vitro fertilization cycles. Here, we demonstrate that the transcription factor Forkhead Box O1 (FOXO1) is a critical regulator of endometrial receptivity in vivo. Uterine ablation of Foxo1 using the progesterone receptor Cre (PgrCre) mouse model resulted in infertility due to altered epithelial cell polarity and apoptosis, preventing the embryo from penetrating the luminal epithelium. Analysis of the uterine transcriptome after Foxo1 ablation identified alterations in gene expression for transcripts involved in the activation of cell invasion, molecular transport, apoptosis, β-catenin (CTNNB1) signaling pathway, and an increase in PGR signaling. The increase of PGR signaling was due to PGR expression being retained in the uterine epithelium during the window of receptivity. Constitutive expression of epithelial PGR during this receptive period inhibited expression of FOXO1 in the nucleus of the uterine epithelium. The reciprocal expression of PGR and FOXO1 was conserved in human endometrial samples during the proliferative and secretory phase. This demonstrates that expression of FOXO1 and the loss of PGR during the window of receptivity are interrelated and critical for embryo implantation.}, number={11}, journal={PLOS GENETICS}, author={Vasquez, Yasmin M. and Wang, Xiaoqiu and Wetendorf, Margeaux and Franco, Heather L. and Mo, Qianxing and Wang, Tianyuan and Lanz, Rainer B. and Young, Steven L. and Lessey, Bruce A. and Spencer, Thomas E. and et al.}, year={2018}, month={Nov} }
 @article{lenis_johnson_wang_tang_dunlap_satterfield_wu_hansen_bazer_2018, title={Functional roles of ornithine decarboxylase and arginine decarboxylase during the peri-implantation period of pregnancy in sheep}, volume={9}, ISSN={2049-1891}, url={http://dx.doi.org/10.1186/s40104-017-0225-x}, DOI={10.1186/s40104-017-0225-x}, abstractNote={Polyamines stimulate DNA transcription and mRNA translation for protein synthesis in trophectoderm cells, as well as proliferation and migration of cells; therefore, they are essential for development and survival of conceptuses (embryo/fetus and placenta). The ovine conceptus produces polyamines via classical and non-classical pathways. In the classical pathway, arginine (Arg) is transformed into ornithine, which is then decarboxylated by ornithine decarboxylase (ODC1) to produce putrescine which is the substrate for the production of spermidine and spermine. In the non-classical pathway, Arg is converted to agmatine (Agm) by arginine decarboxylase (ADC), and Agm is converted to putrescine by agmatinase (AGMAT).Morpholino antisense oligonucleotides (MAOs) were designed and synthesized to inhibit translational initiation of the mRNAs for ODC1 and ADC, in ovine conceptuses.The morphologies of MAO control, MAO-ODC1, and MAO-ADC conceptuses were normal. Double knockdown of ODC1 and ADC (MAO-ODC1:ADC) resulted in two phenotypes of conceptuses; 33% of conceptuses appeared to be morphologically and functionally normal (phenotype a) and 67% of the conceptuses presented an abnormal morphology and functionality (phenotype b). Furthermore, MAO-ODC1:ADC (a) conceptuses had greater tissue concentrations of Agm, putrescine, and spermidine than MAO control conceptuses, while MAO-ODC1:ADC (b) conceptuses only had greater tissue concentrations of Agm . Uterine flushes from ewes with MAO-ODC1:ADC (a) had greater amounts of arginine, aspartate, tyrosine, citrulline, lysine, phenylalanine, isoleucine, leucine, and glutamine, while uterine flushes of ewes with MAO-ODC1:ADC (b) conceptuses had lower amount of putrescine, spermidine, spermine, alanine, aspartate, glutamine, tyrosine, phenylalanine, isoleucine, leucine, and lysine.The double-knockdown of translation of ODC1 and ADC mRNAs was most detrimental to conceptus development and their production of interferon tau (IFNT). Agm, polyamines, amino acids, and adequate secretion of IFNT are critical for establishment and maintenance of pregnancy during the peri-implantation period of gestation in sheep.}, number={1}, journal={Journal of Animal Science and Biotechnology}, publisher={Springer Science and Business Media LLC}, author={Lenis, Yasser Y. and Johnson, Gregory A. and Wang, Xiaoqiu and Tang, Wendy W. and Dunlap, Kathrin A. and Satterfield, M. Carey and Wu, Guoyao and Hansen, Thomas R. and Bazer, Fuller W.}, year={2018}, month={Jan} }
 @article{wang_zhu_feng_lin_wu_li_wang_2018, title={Innate differences and colostrum-induced alterations of jejunal mucosal proteins in piglets with intra-uterine growth restriction}, volume={119}, ISSN={0007-1145 1475-2662}, url={http://dx.doi.org/10.1017/s0007114518000375}, DOI={10.1017/s0007114518000375}, abstractNote={Mammalian neonates undergo rapid transitions from a sterile uterine environment with a continuous intravenous supply of nutrients to a microbe-rich environment with intermittent ingesting of colostrum/milk via the gut. Currently, little is known about the colostrum-induced alterations of intestinal mucosal proteins in piglets with intra-uterine growth restriction (IUGR). In this study, we sought to investigate the innate differences and effects of colostrum on alterations in small-intestinal proteomes of IUGR piglets. Two IUGR (approximately 0·9 kg) and two normal-birth weight (NBW; approximately 1·3 kg) piglets were obtained from each of six sows at birth. One half (n 12; 6 IUGR v. 6 NBW) of the selected newborn piglets were killed to obtain jejunum samples, and the other half (n 12; 6 IUGR v. 6 NBW) of the newborn piglets were allowed to suckle colostrum from their own mothers for 24 h before jejunum sample collection. On the basis of proteomic analysis, we identified thirty-one differentially expressed proteins in the jejunal mucosa between IUGR and normal neonates before or after colostrum consumption. The intestinal proteins altered by colostrum feeding play important roles in the following: (1) increasing intestinal integrity, transport of nutrients, energy metabolism, protein synthesis, immune response and, therefore, cell proliferation; and (2) decreasing oxidative stress, and therefore cell apoptosis, in IUGR neonates. However, colostrum only partially ameliorated the inferior status of the jejunal mucosa in IUGR neonates. These findings provide the first evidence in intestinal protein alterations of IUGR neonates in response to colostrum ingestion, and thus render new insights into the mechanisms responsible for impaired growth in IUGR neonates and into new nutritional intervention strategies.}, number={7}, journal={British Journal of Nutrition}, publisher={Cambridge University Press (CUP)}, author={Wang, Xiaoqiu and Zhu, Yuhua and Feng, Cuiping and Lin, Gang and Wu, Guoyao and Li, Defa and Wang, Junjun}, year={2018}, month={Mar}, pages={734–747} }
 @article{wang_li_wang_wu_jeong_kim_young_lessey_lanz_lydon_et al._2018, title={SOX17 regulates uterine epithelial-stromal cross-talk acting via a distal enhancer upstream of Ihh}, volume={9}, ISSN={["2041-1723"]}, DOI={10.1038/s41467-018-06652-w}, abstractNote={Mammalian pregnancy depends on the ability of the uterus to support embryo implantation. Previous studies reveal the Sox17 gene as a downstream target of the Pgr-Gata2-dependent transcription network that directs genomic actions in the uterine endometrium receptive for embryo implantation. Here, we report that ablating Sox17 in the uterine epithelium impairs leukemia inhibitory factor (LIF) and Indian hedgehog homolog (IHH) signaling, leading to failure of embryo implantation. In vivo deletion of the SOX17-binding region 19 kb upstream of the Ihh locus by CRISPR-Cas technology reduces Ihh expression specifically in the uterus and alters proper endometrial epithelial-stromal interactions, thereby impairing pregnancy. This SOX17-binding interval is also bound by GATA2, FOXA2, and PGR. This cluster of transcription factor binding is common in 737 uterine genes and may represent a key regulatory element essential for uterine epithelial gene expression.}, journal={NATURE COMMUNICATIONS}, author={Wang, Xiaoqiu and Li, Xilong and Wang, Tianyuan and Wu, San-Pin and Jeong, Jae-Wook and Kim, Tae Hoon and Young, Steven L. and Lessey, Bruce A. and Lanz, Rainer B. and Lydon, John P. and et al.}, year={2018}, month={Oct} }
 @article{woods_perez-garcia_kieckbusch_wang_demayo_colucci_hemberger_2017, title={Decidualisation and placentation defects are a major cause of age-related reproductive decline}, volume={8}, ISSN={2041-1723}, url={http://dx.doi.org/10.1038/s41467-017-00308-x}, DOI={10.1038/s41467-017-00308-x}, abstractNote={Mammalian reproductive performance declines rapidly with advanced maternal age. This effect is largely attributed to the exponential increase in chromosome segregation errors in the oocyte with age. Yet many pregnancy complications and birth defects that become more frequent in older mothers, in both humans and mice, occur in the absence of karyotypic abnormalities. Here, we report that abnormal embryonic development in aged female mice is associated with severe placentation defects, which result from major deficits in the decidualisation response of the uterine stroma. This problem is rooted in a blunted hormonal responsiveness of the ageing uterus. Importantly, a young uterine environment can restore normal placental as well as embryonic development. Our data highlight the pivotal, albeit under-appreciated, impact of maternal age on uterine adaptability to pregnancy as major contributor to the decline in reproductive success in older females.Advanced maternal age has been associated with lower reproductive success and higher risk of pregnancy complications. Here the authors show that maternal ageing-related embryonic abnormalities in mouse are caused by decidualisation and placentation defects that can be rescued by transferring the embryo from an old to a young uterus.}, number={1}, journal={Nature Communications}, publisher={Springer Science and Business Media LLC}, author={Woods, Laura and Perez-Garcia, Vicente and Kieckbusch, Jens and Wang, Xiaoqiu and DeMayo, Francesco and Colucci, Francesco and Hemberger, Myriam}, year={2017}, month={Sep} }
 @article{wetendorf_wu_wang_creighton_wang_lanz_blok_tsai_tsai_lydon_et al._2017, title={Decreased epithelial progesterone receptor A at the window of receptivity is required for preparation of the endometrium for embryo attachment†}, volume={96}, ISSN={0006-3363 1529-7268}, url={http://dx.doi.org/10.1095/biolreprod.116.144410}, DOI={10.1095/biolreprod.116.144410}, abstractNote={Abstract 
 The precise timing of progesterone signaling through its cognate receptor, the progesterone receptor (PGR), is critical for the establishment and maintenance of pregnancy. Loss of PGR expression in the murine uterine epithelium during the preimplantation period is a marker for uterine receptivity and embryo attachment. We hypothesized that the decrease in progesterone receptor A (PGRA) expression is necessary for successful embryo implantation. To test this hypothesis, a mouse model constitutively expressing PGRA (mPgrA LsL/+) was generated. Expression of PGRA in all uterine compartments (Pgrcre ) or uterine epithelium (Wnt7acre ) resulted in infertility with defects in embryo attachment and stromal decidualization. Expression of critical PGRA target genes, indian hedgehog, and amphiregulin (Areg), wasmaintained through the window of receptivity while the estrogen receptor target gene, the leukemia inhibitory factor (Lif), a key regulator of embryo receptivity, was decreased. Transcriptomic and cistromic analyses of the mouse uterus at day 4.5 of pregnancy identified an altered group of genes regulating molecular transport in the control of fluid and ion levels within the uterine interstitial space. Additionally, LIF and its cognate receptor, the leukemia inhibitory factor receptor (LIFR), exhibited PGR-binding events in regions upstream of the transcriptional start sites, suggesting PGRA is inhibiting transcription at these loci. Therefore, downregulation of the PGRA isoform at the window of receptivity is necessary for the attenuation of hedgehog signaling, transcriptional activation of LIF signaling, and modulation of solutes and fluid, producing a receptive environment for the attaching embryo. Summary Sentence Expression of PGRA at the window of receptivity transcriptionally represses LIF signaling and aberrantly regulates hedgehog and solute signaling rendering the uterus unreceptive to the implanting embryo.}, number={2}, journal={Biology of Reproduction}, publisher={Oxford University Press (OUP)}, author={Wetendorf, Margeaux and Wu, San-Pin and Wang, Xiaoqiu and Creighton, Chad J. and Wang, Tianyuan and Lanz, Rainer B. and Blok, Leen and Tsai, Sophia Y. and Tsai, Ming-Jer and Lydon, John P. and et al.}, year={2017}, month={Jan}, pages={313–326} }
 @inbook{wang_dunlap_2017, title={Delivery of Morpholino Antisense Oligonucleotides to a Developing Ovine Conceptus via Luminal Injection into a Ligated Uterine Horn}, ISBN={9781493968152 9781493968176}, ISSN={1064-3745 1940-6029}, url={http://dx.doi.org/10.1007/978-1-4939-6817-6_20}, DOI={10.1007/978-1-4939-6817-6_20}, abstractNote={In vivo delivery of morpholino antisense oligonucleotides (MAO) directly into the uterine lumen of a peri-implantation period pregnant sheep is an effective technique for evaluation of gene products for conceptus development. The highly phagocytic conceptus is undergoing rapid morphological change, thereby the available MAO are readily consumed and delivered to developing cells. Here, we describe the method for preparation and surgical delivery of MAO—Endo-Porter complex to developing ovine conceptus on day 8 postmating. Also outlined are methods for posttreatment sample recovery on day 16 postmating.}, booktitle={Morpholino Oligomers}, publisher={Springer New York}, author={Wang, Xiaoqui and Dunlap, Kathrin A.}, year={2017}, pages={241–250} }
 @article{wang_wu_demayo_2017, title={Hormone dependent uterine epithelial-stromal communication for pregnancy support}, volume={60}, ISSN={0143-4004}, url={http://dx.doi.org/10.1016/j.placenta.2017.07.003}, DOI={10.1016/j.placenta.2017.07.003}, abstractNote={Human fertility is a relatively inefficient process. Despite the presence of visibly healthy embryos, 30% of pregnancies generated by assisted reproductive technology (ART) fail before the second trimester. The uterine microenvironment plays a critical role in establishing and maintaining a successful pregnancy that requires coordinated communication between the epithelial and stromal cells of the endometrium. The epithelial cells must cease proliferation and become permissive for the conceptus (embryo and associated extraembryonic membranes), while the stromal cells undergoes mesenchymal-to-epithelioid transformation to form the decidua in support of subsequent embryo development. The ovarian steroids Estrogen (E2) and Progesterone (P4) are the major hormones governing these processes. These hormones act via their nuclear receptors, the estrogen receptor, ESR1, and progesterone receptor, PGR, to direct the transcription of genes that orchestrate epithelial and stromal cell communication. This review will discuss the molecular mechanisms utilized by steroid hormones that regulate uterine receptivity, as well, establish and maintain pregnancy.}, journal={Placenta}, publisher={Elsevier BV}, author={Wang, Xiaoqiu and Wu, San-Pin and DeMayo, Francesco J.}, year={2017}, month={Dec}, pages={S20–S26} }
 @article{lenis_wang_tang_wu_bazer_2016, title={Effects of agmatine on secretion of interferon tau and catecholamines and expression of genes related to production of polyamines by ovine trophectoderm cells}, volume={48}, ISSN={0939-4451 1438-2199}, url={http://dx.doi.org/10.1007/s00726-016-2216-1}, DOI={10.1007/s00726-016-2216-1}, number={10}, journal={Amino Acids}, publisher={Springer Science and Business Media LLC}, author={Lenis, Yasser Y. and Wang, Xiaoqiu and Tang, Wanjin and Wu, Guoyao and Bazer, Fuller W.}, year={2016}, month={Apr}, pages={2389–2399} }
 @article{wang_li_wu_bazer_2016, title={Functional Roles of Fructose: Crosstalk between O-Linked Glycosylation and Phosphorylation of Akt-TSC2-MTOR Cell Signaling Cascade in Ovine Trophectoderm Cells}, volume={95}, ISSN={0006-3363 1529-7268}, url={http://dx.doi.org/10.1095/biolreprod.116.142281}, DOI={10.1095/biolreprod.116.142281}, abstractNote={ABSTRACT During pregnancy, the placentae of ungulate mammals (e.g., cows, sheep, and pigs) convert glucose into fructose, which is the most abundant hexose sugar in fetal fluids and blood. However, the role of fructose, the most enigmatic component of carbohydrate metabolism in fetal-placental tissues, is largely ignored because it is not metabolized via the glycolytic pathway or the Krebs cycle as an energy source. Here we provided evidence for biological functions of fructose that affect proliferative behavior of the conceptus trophectoderm/chorion via activation of the Akt-TSC2-MTOR signaling cascade. The phosphorylation for activation of this cascade is mediated by O-linked glycosylation with UDP-N-acetylglucosamine, a primary product of the hexosamine biosynthesis pathway. These results reveal novel functional roles of fructose in promoting embryonic/fetal growth and development during pregnancy, and also provide new insight into understanding the relationship between excessive fructose intake and metabolic disorders.}, number={5}, journal={Biology of Reproduction}, publisher={Oxford University Press (OUP)}, author={Wang, X. and Li, D. and Wu, G. and Bazer, F. W.}, year={2016}, month={Sep}, pages={102–102} }
 @article{wang_johnson_burghardt_wu_bazer_2016, title={Uterine Histotroph and Conceptus Development. II. Arginine and Secreted Phosphoprotein 1 Cooperatively Stimulate Migration and Adhesion of Ovine Trophectoderm Cells via Focal Adhesion-MTORC2 Mediated Cytoskeleton Reorganization}, volume={95}, ISSN={0006-3363 1529-7268}, url={http://dx.doi.org/10.1095/biolreprod.115.137943}, DOI={10.1095/biolreprod.115.137943}, abstractNote={ABSTRACT In all mammalian species, critical events, including uterine receptivity and development of the conceptus (embryo/fetus and its associated extraembryonic membranes), must be intricately orchestrated and carefully timed during the window of implantation. Otherwise, failure of conceptuses to implant is inevitable, which accounts for 50%–75% of failures to establish pregnancy. Unlike human and rodent blastocysts, the blastocysts of pigs and ruminants undergo rapid transitions from spherical to tubular and filamentous conceptuses in response to histotroph during the peri-implantation period of pregnancy. Both arginine and secreted phosphoprotein 1 (SPP1; also known as osteopontin) are multifunctional molecules that increase significantly in ovine uterine histotroph during early pregnancy; however, little is known about their relationship and synergistic effects on conceptus development. Therefore, we conducted in vitro experiments using our established ovine trophectoderm cell line (oTr1) isolated from Day 15 ovine conceptuses to determine their migratory and adhesive responses to individual and combined effects of arginine and recombinant SPP1 (rSPP1) that contains an Arg-Gly-Asp (RGD) binding sequence. Migration and adhesion of oTr1 cells were significantly stimulated by rSPP1, whereas arginine alone only induced a significant increase in cell migration. However, the combination of arginine and rSPP1 had an additive effect on migration, and a synergistic effect on adhesion of oTr1 cells. Those cooperative effects of arginine and SPP1 were mediated by focal adhesion assembly-MTORC2-cytoskeletal reorganization and MAPK pathways. Collectively, results suggest that arginine and SPP1 in histotroph affect cellular events required for rapid elongation of ovine conceptuses during the peri-implantation period of pregnancy.}, number={3}, journal={Biology of Reproduction}, publisher={Oxford University Press (OUP)}, author={Wang, X. and Johnson, G. A. and Burghardt, R. C. and Wu, G. and Bazer, F. W.}, year={2016}, month={Aug}, pages={71–71} }
 @inbook{wang_wu_bazer_2016, place={Cambridge, MA}, title={mTOR: The Master Regulator of Conceptus Development in Response to Uterine Histotroph During Pregnancy in Ungulates}, ISBN={9780128027332}, DOI={10.1016/B978-0-12-802733-2.00016-5}, abstractNote={Embryonic mortality is a major constraint to reproductive performance in all mammals. Estimates of embryonic death in most mammals, including swine, sheep, cattle, and humans, ranges from 20% to 40%, with two-thirds of the losses occurring during the peri-implantation period of pregnancy. During that stage of pregnancy, the dialog between the mammalian conceptus (embryo/fetus and associated membranes) and maternal uterus involves signaling for pregnancy recognition and maintenance of pregnancy as the stage is set for implantation and placentation that precedes fetal development. Uterine epithelial cells secrete and/or transport a wide range of molecules, including nutrients, collectively referred to as histotroph, that are transported into the fetal–placental vascular system to support growth and development of the conceptus. This review provides a framework for studies of constituents, including hexose sugars (i.e., glucose and fructose), extracellular matrix proteins (e.g., secreted phosphoprotein 1), amino acids (e.g., arginine, leucine, and glutamine), and their metabolites (e.g., nitric oxide and polyamines) that independently and cooperatively activate nutrient-sensing cell signaling pathways, particularly mechanistic target of rapamycin (mTOR), the master regulator of cell growth (proliferative growth via mTORC1 and spatial growth via mTORC2), for growth, development, and survival of conceptuses, as well as for the optimization of culture media for in vitro studies of conceptus development. Understanding the relationships between cell signaling pathways and developmental events is critical for enhancing conceptus development, implantation, and placentation, thereby increasing the probability for birth of healthy offspring.}, booktitle={Molecules to Medicine with mTOR: Translating Critical Pathways Into Novel Therapeutic Strategies}, publisher={Elsevier}, author={Wang, X. and Wu, G. and Bazer, F.W.}, editor={Maiese, K.Editor}, year={2016}, pages={23–35} }
 @article{ruiz-gonzález_xu_wang_burghardt_dunlap_bazer_2015, title={Exosomes, endogenous retroviruses and toll-like receptors: pregnancy recognition in ewes}, volume={149}, ISSN={1470-1626 1741-7899}, url={http://dx.doi.org/10.1530/rep-14-0538}, DOI={10.1530/rep-14-0538}, abstractNote={Conceptus-endometrial communication during the peri-implantation period of pregnancy ensures establishment of pregnancy. We hypothesized that this dialog involves exosomes, ovine endogenous jaagsiekte retroviruses (enJSRV) and toll-like receptors (TLR) which regulate the secretion of interferon tau (IFNT), the pregnancy recognition signal in ruminants. First, exosomes isolated from uterine flushings from cyclic and pregnant ewes were analyzed for exosomal content and uterine expression of heat shock protein 70 (HSC70). Then, conceptus trophectoderm cells (oTr1) treated with different doses of exosomes were analyzed for the expression of genes involved in TLR-mediated cell signaling. The results revealed that exosomes contain mRNAs for enJSRV-ENV, HSC70, interleukins, and interferon (IFN)-regulatory factors. Exosomal content of enJSRV-ENV mRNA and protein decreased from days 10 and 12 to day 16 of gestation, and uterine expression of HSC70 increased in pregnant ewes compared with cyclic ewes. The oTr1 cells proliferated and secreted IFNT in a dose-dependent manner in response to exosomes from cyclic ewes. The expression of CD14, CD68, IRAK1, TRAF6, IRF6, and IRF7 mRNAs that are key to TLR-mediated expression of type 1 IFNs was significantly influenced by day of pregnancy. This study demonstrated that exosomes are liberated into the uterine lumen during the estrous cycle and early pregnancy; however, in pregnant ewes, exosomes stimulate trophectoderm cells to proliferate and secrete IFNT coordinately with regulation of TLR-mediated cell signaling. These results support our hypothesis that free and/or exosomal enJSRV act on the trophectoderm via TLR to induce the secretion of IFNT in a manner similar to that for innate immune responses of macrophages and plasmacytoid dendritic cells to viral pathogens.}, number={3}, journal={REPRODUCTION}, publisher={Bioscientifica}, author={Ruiz-González, Irene and Xu, Jing and Wang, Xiaoqiu and Burghardt, Robert C and Dunlap, Kathrin A and Bazer, Fuller W}, year={2015}, month={Mar}, pages={281–291} }
 @article{wang_burghardt_romero_hansen_wu_bazer_2015, title={Functional Roles of Arginine During the Peri-Implantation Period of Pregnancy. III. Arginine Stimulates Proliferation and Interferon Tau Production by Ovine Trophectoderm Cells via Nitric Oxide and Polyamine-TSC2-MTOR Signaling Pathways}, volume={92}, ISSN={0006-3363 1529-7268}, url={http://dx.doi.org/10.1095/biolreprod.114.125989}, DOI={10.1095/biolreprod.114.125989}, abstractNote={ABSTRACT In mammal species, arginine is a multifunctional amino acid required for survival, growth, and development of conceptuses (embryo/fetus and associated extraembryonic membranes) during the peri-implantation period of pregnancy. However, functional roles of arginine with respect to it being a substrate for production of nitric oxide (NO) and polyamines on trophectoderm cell proliferation and function remain largely unknown. To systematically assess roles of arginine in conceptus development and its effect on interferon tau (IFNT) production for pregnancy recognition signaling in ruminants, an established ovine trophectoderm (oTr1) cell line isolated from Day-15 ovine conceptuses were used to determine their response to arginine, putrescine, and NO donors, as well as their associated inhibitors. Arginine at physiological concentration (0.2 mM) stimulated maximum oTr cell proliferation (increased 2.0-fold at 48 h and 2.6-fold at 96 h; P < 0.05), stimulated IFNT production (IFNT/cell increased 3.1-fold; P < 0.05), and increased total protein per cell by more than 1.5-fold (P < 0.05). It also increased phosphorylated tuberous sclerosis protein (p-TSC2) and phosphorylated mechanistic target of rapamycin (MTOR) abundance by more than 2.7- and 4.3-fold (P < 0.0001) after long-term incubation, respectively. When Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME; NO synthase inhibitor), DL-α-difluoromethylornithine hydrochloride hydrate (DFMO; ornithine decarboxylase inhibitor), and the combination (L-NAME + DFMO) were added, the effects of arginine on cell proliferation was reduced by 10.7%, 16.1%, and 22.3% (P < 0.05) at 48 h, and 15.3%, 27.2%, and 39.1% (P < 0.05) at 96 h of incubation, respectively, but values remained 1.5-fold higher (P < 0.05) than for the arginine-free control, which suggests that arginine, per se, serves as a growth factor. Both putrescine and NO stimulate cell proliferation via activation of the TSC2-MTOR signaling cascade, whereas only putrescine increased IFNT production. Collectively, our results indicate that arginine is essential for oTr1 cell proliferation and IFNT production via the NO/polyamine-TSC2-MTOR signaling pathways, particularly the pathway involving polyamine biosynthesis.}, number={3}, journal={Biology of Reproduction}, publisher={Oxford University Press (OUP)}, author={Wang, Xiaoqiu and Burghardt, Robert C. and Romero, Jared J. and Hansen, Thomas R. and Wu, Guoyao and Bazer, Fuller W.}, year={2015}, month={Mar} }
 @article{ruiz-gonzález_minten_wang_dunlap_bazer_2015, title={Involvement of TLR7 and TLR8 in conceptus development and establishment of pregnancy in sheep}, volume={149}, ISSN={1470-1626 1741-7899}, url={http://dx.doi.org/10.1530/rep-14-0537}, DOI={10.1530/rep-14-0537}, abstractNote={Toll-like receptors (TLRs) belong to the innate immune system and regulate inflammatory events that affect mammalian reproduction. In Study 1, we demonstrated that abundance of ovine TLR1-TLR9 mRNAs in the uterus differs due to reproductive status (TLR2, TLR3, TLR7, and TLR8) and the day of the estrous cycle and pregnancy (TLR1-TLR3, TLR5-TLR7, and TLR9). Expression of TLR7 and TLR8 proteins was localized primarily to uterine epithelia and stroma and regulated in a temporal manner. In Study 2, we determined that ovine conceptuses express TLR7 and TLR8 on all days studied and that expression of the envelope protein of ovine endogenous retrovirus (enJSRV-Env) declined in conceptus trophectoderm from Day 13 to Day 16 of pregnancy. In Study 3, loss-of-function experiments were conducted in vivo using morpholino antisense oligonucleotides (MAOs) injected into the uterine lumen to block synthesis of TLR7 and TLR8 proteins, individually and jointly. Conceptuses were recovered on Day 16 to assess their morphology. MAO-treated conceptuses were developmentally retarded, produced less interferon tau (IFNT), and had fewer binucleate cells (BNCs) compared with MAO-Controls. Moreover, expression of enJSRV-Env mRNA in MAO-TLR7 conceptuses was greater than that for MAO-Control and MAO-TLR8 conceptuses, but similar to MAO-TLR7/TLR8 conceptuses. Results of this study indicated differences in TLR1-TLR9 expression due to reproductive status and the day of the estrous cycle and pregnancy. TLR7 and TLR8 also influence development, enJSRV-Env abundance, secretion of IFNT, and formation of BNCs by conceptuses. These findings corroborate our hypothesis that TLR7 and TLR8 mediate pathways whereby enJSRV-Env regulates key peri-implantation events in conceptus development and differentiated functions of trophectoderm cells.}, number={4}, journal={REPRODUCTION}, publisher={Bioscientifica}, author={Ruiz-González, Irene and Minten, Megan and Wang, Xiaoqiu and Dunlap, Kathrin A and Bazer, Fuller W}, year={2015}, month={Apr}, pages={305–316} }
 @article{bazer_wang_johnson_wu_2015, title={Select nutrients and their effects on conceptus development in mammals}, volume={1}, ISSN={2405-6545}, url={http://dx.doi.org/10.1016/j.aninu.2015.07.005}, DOI={10.1016/j.aninu.2015.07.005}, abstractNote={The dialogue between the mammalian conceptus (embryo/fetus and associated membranes) involves signaling for pregnancy recognition and maintenance of pregnancy during the critical peri-implantation period of pregnancy when the stage is set for implantation and placentation that precedes fetal development. Uterine epithelial cells secrete and/or transport a wide range of molecules, including nutrients, collectively referred to as histotroph that are transported into the fetal-placental vascular system to support growth and development of the conceptus. The availability of uterine-derived histotroph has long-term consequences for the health and well-being of the fetus and the prevention of adult onset of metabolic diseases. Histotroph includes numerous amino acids, but arginine plays a particularly important role as a source of nitric oxide and polyamines required for fetal-placental development in rodents, swine and humans through mechanisms that remain to be fully elucidated. Mechanisms whereby arginine regulates expression of genes via the mechanistic target of rapamycin cell signaling pathways critical to conceptus development, implantation and placentation are discussed in detail in this review.}, number={3}, journal={Animal Nutrition}, publisher={Elsevier BV}, author={Bazer, Fuller W. and Wang, Xiaoqiu and Johnson, Greg A. and Wu, Guoyao}, year={2015}, month={Sep}, pages={85–95} }
 @article{wu_bazer_wang_johnson_hou_dai_wang_wu_2015, title={Synthesis of polyamines from L-proline in the porcine placenta and neonatal enterocytes}, volume={47}, number={8}, journal={Amino Acids}, author={Wu, G. and Bazer, F.W. and Wang, X. and Johnson, G.A. and Hou, Y. and Dai, Z. and Wang, J. and Wu, Z.}, year={2015}, pages={1669} }
 @article{bazer_ying_wang_dunlap_zhou_johnson_wu_2015, title={The many faces of interferon tau}, volume={47}, ISSN={0939-4451 1438-2199}, url={http://dx.doi.org/10.1007/s00726-014-1905-x}, DOI={10.1007/s00726-014-1905-x}, number={3}, journal={Amino Acids}, publisher={Springer Science and Business Media LLC}, author={Bazer, Fuller W. and Ying, Wei and Wang, Xiaoqiu and Dunlap, Kathrin A. and Zhou, Beiyan and Johnson, Greg A. and Wu, Guoyao}, year={2015}, month={Jan}, pages={449–460} }
 @article{bazer_ruiz-gonzalez_xu_wang_dunlap_2015, title={The many faces of interferon tau at the maternal-conceptus interface}, volume={73}, number={S2}, journal={American Journal of Reproductive Immunology}, author={Bazer, F.W. and Ruiz-Gonzalez, I. and Xu, J. and Wang, X. and Dunlap, K.A.}, year={2015}, pages={21} }
 @article{wang_johnson_burghardt_wu_bazer_2015, title={Uterine Histotroph and Conceptus Development. I. Cooperative Effects of Arginine and Secreted Phosphoprotein 1 on Proliferation of Ovine Trophectoderm Cells via Activation of the PDK1-Akt/PKB-TSC2-MTORC1 Signaling Cascade}, volume={92}, ISSN={0006-3363 1529-7268}, url={http://dx.doi.org/10.1095/biolreprod.114.125971}, DOI={10.1095/biolreprod.114.125971}, abstractNote={ABSTRACT The greatest limitation to reproductive performance in most mammals, including humans, is embryonic mortality, which, in general, claims 20%–40% of the embryos during the peri-implantation period of pregnancy. Both arginine and secreted phosphoprotein 1 (SPP1) are multifunctional molecules that increase significantly in ovine uterine histotroph during early pregnancy. However, little is known about the relationship and underlying mechanisms for synergistic effects of arginine and SPP1, if any, on conceptus (embryo/fetus and associated extraembryonic membranes) development. Therefore, we conducted in vitro experiments using our established ovine trophectoderm cell line (oTr1) isolated from Day 15 ovine conceptuses to determine their proliferative response to individual and synergistic effects of arginine and recombinant SPP1 (rSPP1) that contains an RGD binding sequence. At physiological concentrations, arginine (0.2 mM) stimulated oTr1 cell proliferation 1.7-fold (P < 0.05) at 48 h, whereas rSPP1 (10 ng/ml) had no such effect. However, an additive effect on oTr1 cell proliferation was induced by combination of arginine and SPP1 as compared to the control (2.1-fold increase; P < 0.01), arginine alone (1.3-fold increase; P < 0.05), and rSPP1 alone (1.5-fold increase; P < 0.01). This additive effect was mediated through cooperative activation of the PDK1-Akt/PKB-TSC2-MTORC1 cell signaling cascade. Collectively, results suggest that arginine and SPP1 in histotroph act cooperatively to enhance survival, growth, and development of ovine conceptuses.}, number={2}, journal={Biology of Reproduction}, publisher={Oxford University Press (OUP)}, author={Wang, Xiaoqiu and Johnson, Greg A. and Burghardt, Robert C. and Wu, Guoyao and Bazer, Fuller W.}, year={2015}, month={Feb} }
 @article{dai_wu_wang_wang_jia_bazer_wu_2014, title={Analysis of polyamines in biological samples by HPLC involving pre-column derivatization with o-phthalaldehyde and N-acetyl-l-cysteine}, volume={46}, ISSN={0939-4451 1438-2199}, url={http://dx.doi.org/10.1007/s00726-014-1717-z}, DOI={10.1007/s00726-014-1717-z}, number={6}, journal={Amino Acids}, publisher={Springer Science and Business Media LLC}, author={Dai, Zhaolai and Wu, Zhenlong and Wang, Junjun and Wang, Xiaoqiu and Jia, Sichao and Bazer, Fuller W. and Wu, Guoyao}, year={2014}, month={Mar}, pages={1557–1564} }
 @article{wang_ying_dunlap_lin_satterfield_burghardt_wu_bazer_2014, title={Arginine Decarboxylase and Agmatinase: An Alternative Pathway for De Novo Biosynthesis of Polyamines for Development of Mammalian Conceptuses}, volume={90}, ISSN={0006-3363 1529-7268}, url={http://dx.doi.org/10.1095/biolreprod.113.114637}, DOI={10.1095/biolreprod.113.114637}, abstractNote={ABSTRACT Ornithine decarboxylase (ODC1) is considered the rate-controlling enzyme for the classical de novo biosynthesis of polyamines (putrescine, spermidine, and spermine) in mammals. However, metabolism of arginine to agmatine via arginine decarboxylase (ADC) and conversion of agmatine to polyamines via agmatinase (AGMAT) is an alternative pathway long recognized in lower organisms, but only recently suggested for neurons and liver cells of mammals. We now provide evidence for a functional ADC/AGMAT pathway for the synthesis of polyamines in mammalian reproductive tissue for embryonic survival and development. We first investigated cellular functions of polyamines by in vivo knockdown of translation of mRNA for ODC1 in ovine conceptus trophectoderm using morpholino antisense oligonucleotides (MAOs) and found that one-half of the conceptuses were morphologically and functionally either normal or abnormal. Furthermore, we found that increases in ADC/AGMAT mRNA levels and in the translation of AGMAT mRNA among conceptuses in MAO-ODC1 knockdown compensated for the loss of ODC1, supporting polyamine synthesis from arginine and accounting for the normal and abnormal phenotypes of conceptuses. We conclude that the majority of polyamine synthesis is by the conventional ODC1-dependent pathway (arginine-ornithine-putrescine) and that deficiencies in ODC1 result in increased activity of the rescue ADC/AGMAT-dependent pathway (arginine-agmatine-putrescine) for production of polyamines. The presence of an alternative ADC/AGMAT pathway for converting arginine into putrescine is functionally important for supporting survival and development of mammalian conceptuses.}, number={4}, journal={Biology of Reproduction}, publisher={Oxford University Press (OUP)}, author={Wang, Xiaoqiu and Ying, Wei and Dunlap, Kathrin A. and Lin, Gang and Satterfield, M. Carey and Burghardt, Robert C. and Wu, Guoyao and Bazer, Fuller W.}, year={2014}, month={Apr} }
 @article{bazer_wu_johnson_wang_2014, title={Environmental factors affecting pregnancy: Endocrine disrupters, nutrients and metabolic pathways}, volume={398}, ISSN={0303-7207}, url={http://dx.doi.org/10.1016/j.mce.2014.09.007}, DOI={10.1016/j.mce.2014.09.007}, abstractNote={Uterine adenogenesis, a unique post-natal event in mammals, is vulnerable to endocrine disruption by estrogens and progestins resulting in infertility or reduced prolificacy. The absence of uterine glands results in insufficient transport of nutrients into the uterine lumen to support conceptus development. Arginine, a component of histotroph, is substrate for production of nitric oxide, polyamines and agmatine and, with secreted phosphoprotein 1, it affects cytoskeletal organization of trophectoderm. Arginine is critical for development of the conceptus, pregnancy recognition signaling, implantation and placentation. Conceptuses of ungulates and cetaceans convert glucose to fructose which is metabolized via multiple pathways to support growth and development. However, high fructose corn syrup in soft drinks and foods may increase risks for metabolic disorders and increase insulin resistance in adults. Understanding endocrine disrupters and dietary substances, and novel pathways for nutrient metabolism during pregnancy can improve survival and growth, and prevent chronic metabolic diseases in offspring.}, number={1-2}, journal={Molecular and Cellular Endocrinology}, publisher={Elsevier BV}, author={Bazer, Fuller W. and Wu, Guoyao and Johnson, Gregory A. and Wang, Xiaoqiu}, year={2014}, month={Dec}, pages={53–68} }
 @article{wang_frank_xu_dunlap_satterfield_burghardt_romero_hansen_wu_bazer_2014, title={Functional Role of Arginine During the Peri-implantation Period of Pregnancy. II. Consequences of Loss of Function of Nitric Oxide Synthase NOS3 mRNA in Ovine Conceptus Trophectoderm}, volume={91}, ISSN={0006-3363 1529-7268}, url={http://dx.doi.org/10.1095/biolreprod.114.121202}, DOI={10.1095/biolreprod.114.121202}, abstractNote={ABSTRACT Nitric oxide (NO) is a gaseous molecule that regulates angiogenesis and vasodilation via activation of the cGMP pathway. However, functional roles of NO during embryonic development from spherical blastocysts to elongated filamentous conceptuses (embryo and extraembryonic membrane) during the peri-implantation period of pregnancy have not been elucidated in vivo. In order to assess roles of NO production in survival and development of the ovine conceptus, we conducted an in vivo morpholino antisense oligonucleotide (MAO)-mediated knockdown trial of nitric oxide synthase-3 (NOS3) mRNA, the major isoform of NO synthase, in ovine conceptus trophectoderm (Tr). Translational knockdown of NOS3 mRNA results in small, thin, and underdeveloped conceptuses, but normal production of interferon-tau, the pregnancy recognition signal in sheep. MAO-NOS3 knockdown in conceptuses decreased the abundance of NOS3 (72%, P < 0.05) and the arginine transporter SLC7A1 proteins in conceptus Tr. Furthermore, the amounts of ornithine and polyamines were less (P < 0.01) in uterine fluid, whereas the amounts of arginine (58%, P < 0.01), citrulline (68%, P < 0.05), ornithine (68%, P < 0.001), glutamine (78%, P < 0.001), glutamate (68%, P < 0.05), and polyamines (P < 0.01) were less in conceptuses, which likely accounts for the failure of MAO-NOS3 conceptuses to develop normally. For MAO-NOS3 conceptuses, there were no compensatory increases in the expression levels of either nitric oxide synthase-1 (NOS1) or nitric oxide synthase-2 (NOS2) or in expression of enzymes for synthesis of polyamines (ornithine decarboxylase, arginine decarboxylase, agmatinase) from arginine or ornithine with which to rescue development of MAO-NOS3 conceptuses. Thus, the adverse effect of MAO-NOS3 to reduce NO generation and the transport of arginine and ornithine into conceptuses is central to an explanation for failure of normal development of MAO-NOS3, compared to control conceptuses. The study, for the first time, created an NO-deficient mammalian conceptus model in vivo and provided new insights into the orchestrated events of conceptus development during the peri-implantation period of pregnancy. Our data suggest that NOS3 is the key enzyme for NO production by conceptus Tr and that this protein also regulates the availability of arginine in conceptus tissues for synthesis of polyamines that are essential for conceptus survival and development.}, number={3}, journal={Biology of Reproduction}, publisher={Oxford University Press (OUP)}, author={Wang, Xiaoqiu and Frank, James W. and Xu, Jing and Dunlap, Kathrin A. and Satterfield, M. Carey and Burghardt, Robert C. and Romero, Jared J. and Hansen, Thomas R. and Wu, Guoyao and Bazer, Fuller W.}, year={2014}, month={Sep} }
 @article{wang_frank_little_dunlap_satterfield_burghardt_hansen_wu_bazer_2014, title={Functional role of arginine during the peri‐implantation period of pregnancy. I. Consequences of loss of function of arginine transporter
            SLC7A1
            mRNA in ovine conceptus trophectoderm}, volume={28}, ISSN={0892-6638 1530-6860}, url={http://dx.doi.org/10.1096/fj.13-248757}, DOI={10.1096/fj.13-248757}, abstractNote={Arginine, the common substrate for production of nitric oxide (NO) and polyamines in mammals, increases in the uterine lumen during the peri‐implantation period of pregnancy. However, functional roles of arginine within the uterine lumen for conceptus (embryo and extraembryonic membranes) development have not been elucidated in vivo. To assess roles of arginine in reproductive tissue for survival and development of the conceptus, we conducted an in vivo morpholino antisense oligonucleotide (MAO)‐mediated knockdown of SLC7A1 mRNA, the arginine transporter in ovine conceptus trophectoderm (Tr). Translational knockdown of SLC7A1 mRNA resulted in retarded conceptus development and abnormal function compared to MAO control. Use of MAO‐SLC7A1 knockdown in conceptuses decreased arginine transport (73%, P<0.01), the abundance of ornithine decarboxylase, and nitric oxide synthase (NOS3) proteins, arginine‐related amino acids [citrulline (76%, P<0.05) and ornithine (40%, P<0.05)], and polyamines, which likely accounts for their retarded development. Also, no alternative arginine precursors (glutamine and glutamate), isoforms of nitric oxide synthase (NOS1 and NOS2), or alternative pathways for polyamine biosynthesis via arginine decarboxylase and agmatinase were activated to rescue conceptus development. Collectively, SLC7A1 is the key transporter of arginine by conceptus Tr, and arginine is essential for conceptus survival and development.—Wang, X., Frank, J. W., Little, D. R., Dunlap, K. A., Satterfield, M. C., Burghardt, R. C., Hansen, T. R., Wu, G., and Bazer, F. W. Functional role of arginine during the peri‐implantation period of pregnancy. I. Consequences of loss of function of arginine transporter SLC7A1 mRNA in ovine conceptus trophectoderm. FASEB J. 28, 2852–2863 (2014). www.fasebj.org}, number={7}, journal={The FASEB Journal}, publisher={Wiley}, author={Wang, Xiaoqiu and Frank, James W. and Little, Danielle R. and Dunlap, Kathrin A. and Satterfield, M. Carey and Burghardt, Robert C. and Hansen, Thomas R. and Wu, Guoyao and Bazer, Fuller W.}, year={2014}, month={Mar}, pages={2852–2863} }
 @article{lin_wang_wu_feng_zhou_li_wang_2014, title={Improving amino acid nutrition to prevent intrauterine growth restriction in mammals}, volume={46}, ISSN={0939-4451 1438-2199}, url={http://dx.doi.org/10.1007/s00726-014-1725-z}, DOI={10.1007/s00726-014-1725-z}, number={7}, journal={Amino Acids}, publisher={Springer Science and Business Media LLC}, author={Lin, Gang and Wang, Xiaoqiu and Wu, Guoyao and Feng, Cuiping and Zhou, Huaijun and Li, Defa and Wang, Junjun}, year={2014}, month={Mar}, pages={1605–1623} }
 @article{kong_wang_yin_li_gao_bazer_wu_2014, title={Putrescine Stimulates the mTOR Signaling Pathway and Protein Synthesis in Porcine Trophectoderm Cells}, volume={91}, ISSN={0006-3363 1529-7268}, url={http://dx.doi.org/10.1095/biolreprod.113.113977}, DOI={10.1095/biolreprod.113.113977}, abstractNote={ABSTRACT Insufficient placental growth is a major factor contributing to intrauterine growth retardation in mammals. There is growing evidence that putrescine produced from arginine (Arg) and proline via ornithine decarboxylase is a key regulator of angiogenesis, embryogenesis, as well as placental and fetal growth. However, the underlying mechanisms are largely unknown. The present study tested the hypothesis that putrescine stimulates protein synthesis by activating the mechanistic target of rapamycin (mTOR) signaling pathway in porcine trophectoderm cell line 2 cells. The cells were cultured for 2 to 4 days in customized Arg-free Dulbecco modified Eagle Ham medium containing 0, 10, 25, or 50 μM putrescine or 100 μM Arg. Cell proliferation, protein synthesis, and degradation, as well as the abundance of total and phosphorylated mTOR, ribosomal protein S6 kinase 1, and eukaryotic initiation factor 4E-binding protein-1 (4EBP1), were determined. Our results indicate that putrescine promotes cell proliferation and protein synthesis in a dose- and time-dependent manner, which was inhibited by difluoro-methylornithine (an inhibitor of ornithine decarboxylase). Moreover, supplementation of culture medium with putrescine increased the abundance of phosphorylated mTOR and its downstream targets, 4EBP1 and p70 S6K1 proteins. Collectively, these findings reveal a novel and important role for putrescine in regulating the mTOR signaling pathway in porcine placental cells. We suggest that dietary supplementation with or intravenous administration of putrescine may provide a new and effective strategy to improve survival and growth of embryos/fetuses in mammals.}, number={5}, journal={Biology of Reproduction}, publisher={Oxford University Press (OUP)}, author={Kong, Xiangfeng and Wang, Xiaoqiu and Yin, Yulong and Li, Xilong and Gao, Haijun and Bazer, Fuller W. and Wu, Guoyao}, year={2014}, month={Nov} }
 @article{wang_lin_liu_feng_zhou_wang_li_wu_wang_2014, title={Temporal proteomic analysis reveals defects in small-intestinal development of porcine fetuses with intrauterine growth restriction}, volume={25}, ISSN={0955-2863}, url={http://dx.doi.org/10.1016/j.jnutbio.2014.03.008}, DOI={10.1016/j.jnutbio.2014.03.008}, abstractNote={The fetus/neonate with intrauterine growth restriction (IUGR) has a high perinatal mortality and morbidity rate, as well as reduced efficiency for nutrients utilization. Our previous studies showed alterations of intestinal proteome in IUGR piglets both at birth and during the nursing period. Considering the potential long-term impacts of fetal programming and substantial increases in amounts of amniotic fluid nutrients from mid-gestation in pigs, the present study involved IUGR porcine fetuses from days 60 to 110 of gestation (mid to late gestation). We identified 59 differentially expressed proteins in the fetal small intestine that are related to intestinal growth, development and reprogramming. Our results further indicated increased abundances of proteins and enzymes associated with oxidative stress, apoptosis and protein degradation, as well as decreased abundances of proteins that are required for maintenance of cell structure and motility, absorption and transport of nutrients, energy metabolism, and protein synthesis in the fetal gut. Moreover, IUGR from middle to late gestation was associated with reduced expression of intestinal proteins that participate in regulation of gene expression and signal transduction. Collectively, these findings provide the first evidence for altered proteomes in the small intestine of IUGR fetuses, thereby predisposing the gut to metabolic defects during gestation and neonatal periods.}, number={7}, journal={The Journal of Nutritional Biochemistry}, publisher={Elsevier BV}, author={Wang, Xiaoqiu and Lin, Gang and Liu, Chuang and Feng, Cuiping and Zhou, Huaijun and Wang, Taiji and Li, Defa and Wu, Guoyao and Wang, Junjun}, year={2014}, month={Jul}, pages={785–795} }
 @article{wang_liu_feng_wang_lin_zhu_yin_li_wang_2013, title={IUGR alters muscle fiber development and proteome in fetal pigs }, volume={18}, ISSN={1093-9946 1093-4715}, url={http://dx.doi.org/10.2741/4123}, DOI={10.2741/4123}, abstractNote={Intrauterine growth restriction (IUGR) may have permanent stunting effects on muscle growth and development of the progeny. However, underlying mechanisms are largely unknown. Recent studies comparing muscle fiber development and proteomes in IUGR and normal-body-weight (NBW) fetal pigs indicated that muscle fiber diameter were smaller in IUGR fetal pigs than in NBW fetal pigs on all three stages (d 60, d 90 and d 110) of gestation. Although the number of primary fibers did not differ between these two fetal groups on d 60 of gestation, the total number of muscle fibers in IUGR fetal pigs was lower on d 90 and 110 of gestation, when compared with NBW fetal pigs. Further proteomic analysis has shown that 37 proteins involved in energy supply and protein metabolism, structure and type of muscle fibers, proliferation and differentiation of muscle fibers, nutrient transport, intracellular environment, and tissue integrity were differentially expressed between IUGR and NBW fetal pigs. These novel findings provide some implications on the mechanisms of reduced growth and impaired development of skeletal muscle in IUGR piglets.}, number={2}, journal={Frontiers in Bioscience}, publisher={Frontiers in Bioscience}, author={Wang, Taji and Liu, Chuang and Feng, Cuiping and Wang, Xiaoqiu and Lin, Gang and Zhu, Yuhua and Yin, Jingdong and Li, Defa and Wang, Junjun}, year={2013}, pages={598–607} }
 @article{wu_bazer_satterfield_li_wang_johnson_burghardt_dai_wang_wu_2013, title={Impacts of arginine nutrition on embryonic and fetal development in mammals}, volume={45}, ISSN={0939-4451 1438-2199}, url={http://dx.doi.org/10.1007/s00726-013-1515-z}, DOI={10.1007/s00726-013-1515-z}, number={2}, journal={Amino Acids}, publisher={Springer Science and Business Media LLC}, author={Wu, Guoyao and Bazer, Fuller W. and Satterfield, M. Carey and Li, Xilong and Wang, Xiaoqiu and Johnson, Gregory A. and Burghardt, Robert C. and Dai, Zhaolai and Wang, Junjun and Wu, Zhenlong}, year={2013}, month={Jun}, pages={241–256} }
 @article{liu_lin_wang_wang_wu_li_wang_2013, title={Intrauterine growth restriction alters the hepatic proteome in fetal pigs}, volume={24}, ISSN={0955-2863}, url={http://dx.doi.org/10.1016/j.jnutbio.2012.06.016}, DOI={10.1016/j.jnutbio.2012.06.016}, abstractNote={Intrauterine growth restriction (IUGR) is a major problem in both humans and animals. The IUGR fetus has abnormal metabolism of nutrients in the liver. This study was conducted with comparative proteomic approach and biochemical analyses to test the hypothesis that IUGR alters the hepatic proteome in the fetal liver. Livers were obtained from IUGR and normal-weight fetal pigs at Day 110 of gestation. Twenty-two differentially expressed proteins in the liver were identified between IUGR and normal fetal pigs. These proteins participate in the intermediary metabolism of nutrients (including glucose, amino acids, protein, lipids, vitamins and minerals), oxidative stress, as well as cell structure and growth. Of particular interest, the IUGR fetus had a higher activity of glutamate oxaloacetate transaminase and a lower activity of lipoprotein lipase than the normal ones. These results indicate altered metabolism of nutrients, abnormal ammonia utilization, and reduced capacity for detoxification in the liver of IUGR fetus. Collectively, the findings have important implication for explaining low food efficiency and understanding the mechanism responsible for impaired growth in IUGR neonates.}, number={6}, journal={The Journal of Nutritional Biochemistry}, publisher={Elsevier BV}, author={Liu, Chuang and Lin, Gang and Wang, Xiaoqiu and Wang, Taiji and Wu, Guoyao and Li, Defa and Wang, Junjun}, year={2013}, month={Jun}, pages={954–959} }
 @article{lin_wang_wang_wu_lai_2013, title={T Cells Development Is Different between Thymus from Normal and Intrauterine Growth Restricted Pig Fetus at Different Gestational Stage}, volume={26}, ISSN={1011-2367 1976-5517}, url={http://dx.doi.org/10.5713/ajas.2012.12132}, DOI={10.5713/ajas.2012.12132}, abstractNote={This experiment was conducted to evaluate the development of T cells in intrauterine growth retarded (IUGR) piglets at different gestational stages, and tentatively explore the relationship between T cells development and the Notch signaling pathway. A total of 18 crossbred (Landrace×Large white) primiparous sows were mated at similar weights and estruses and euthanized at d 60, 90 and 110 of gestation with six replicates for each time point. One IUGR and one normal fetus were picked from each litter. The T-cell subsets, mRNA expression of Delta-like1, Delta-like4, Jagged1, and Notch2 genes in the thymus were investigated. Compared to normal piglets, CD3+CD4−CD8+ cells in IUGR fetuses at d 90 was 0.13% lower (p<0.05). At d 110 of gestation CD8+ T cells in IUGR fetuses was 0.19% lower (p<0.05). The percentage of CD8+ T cells was 3.14% lower (p<0.05) of the total T cells in IUGR pigs at d 60. The abundance of Notch2 and Delta-like4 mRNA at d 110 was 20.93% higher and 0.77% (p<0.05) lower, and Delta-like1 mRNA at d 90 was 0.19% (p<0.05) higher compared to normal pigs. These results suggested that normal fetuses had a greater proportion of T-cell subsets at earlier gestation periods, and the Notch signaling pathway was likely partially responsible for these differences to some degree.}, number={3}, journal={Asian-Australasian Journal of Animal Sciences}, publisher={Asian Australasian Association of Animal Production Societies}, author={Lin, Yan and Wang, Junjun and Wang, Xiaoqiu and Wu, Weizong and Lai, Changhua}, year={2013}, month={Mar}, pages={343–348} }
 @article{wang_yang_liu_zhou_wu_qiao_li_wang_2011, title={Dietary Supplementation with the Probiotic Lactobacillus fermentum I5007 and the Antibiotic Aureomycin Differentially Affects the Small Intestinal Proteomes of Weanling Piglets}, volume={142}, ISSN={0022-3166 1541-6100}, url={http://dx.doi.org/10.3945/jn.111.147074}, DOI={10.3945/jn.111.147074}, abstractNote={Antibiotics have long been used in animal production and medication to alleviate weaning stress. However, due to the concerns over food safety and human health, its use in animal production has been prohibited in many countries. Therefore, there is growing interest in developing alternative additives, such as a probiotic Lactobacillus. In this study, a proteomic approach coupled with biochemical analysis was applied to investigate alterations of proteomes in the small intestinal mucosa of weanling piglets after a 13-d period of feeding with supplemental L. fermentum I5007 or aureomycin (an antibiotic). We indentified 27 differentially expressed protein spots that participated in 7 key biological processes, including: 1) energy metabolism; 2) lipid metabolism; 3) protein synthesis; 4) cell structure and mobility; 5) cellular proliferation and apoptosis; 6) immune response; and 7) stress response and detoxification. Both L. fermentum I5007 and aureomycin decreased the expression of proteins related to apoptosis, stress response, and increased the expression of proteins related to detoxification in the gastrointestinal (GI) tract of weanling piglets. L. fermentum I5007 exhibited additional effects in alleviating weaning stress syndrome by enhancing the levels of proteins involved in energy metabolism, lipid metabolism, cell structure and mobility, protein synthesis, and immune response, thereby facilitating cellular proliferation and depressing apoptosis. In contrast, aureomycin reduced the levels of proteins related to energy metabolism, protein synthesis, cell structure, motility, and immunity. These novel findings have important implications for understanding the mechanisms whereby L. fermentum I5007 can improve the GI health of postweaning piglets.}, number={1}, journal={The Journal of Nutrition}, publisher={Oxford University Press (OUP)}, author={Wang, Xiaoqiu and Yang, Fang and Liu, Chuang and Zhou, Huaijun and Wu, Guoyao and Qiao, Shiyan and Li, Defa and Wang, Junjun}, year={2011}, month={Nov}, pages={7–13} }
 @article{wu_wang_wu_kim_chen_wang_2010, title={Differential composition of proteomes in sow colostrum and milk from anterior and posterior mammary glands1}, volume={88}, ISSN={0021-8812 1525-3163}, url={http://dx.doi.org/10.2527/jas.2010-2972}, DOI={10.2527/jas.2010-2972}, abstractNote={Piglets obtaining milk from anterior and middle mammary glands (MG) grow faster than those suckling posterior MG, but the underlying mechanisms are not clear. The purpose of this study was to investigate the differential proteomes of colostrum and milk secreted by anterior and posterior MG. Six healthy primiparous sows with 7 pairs of MG were used; the first and the second pairs were defined as anterior MG and the sixth and seventh pairs as posterior MG. Colostrum and milk were collected at d 1 and 14 after parturition, respectively. Comparative proteomics analysis was performed to identify the differentially expressed proteins in colostrum and milk secreted by anterior and posterior MG. Results show that protein composition in colostrum and milk varied markedly with the anatomical location of MG. Immunoglobulins, lactadherin, and haptoglobin were upregulated (P < 0.05) in colostrum from anterior MG compared with posterior MG. Concentrations of immunoglobulins and lactoferrin in milk from anterior MG were greater (P < 0.05) than milk from posterior MG. Moreover, concentration of proteins from somatic cells was greater (P < 0.05) in milk from posterior MG compared with anterior MG. Most proteins, in which abundance was upregulated in colostrum and milk from anterior MG, contribute to passive immunity, intestinal development of suckling piglets and epithelial integrity, and the health of MG. Collectively, these results indicate that in comparison with posterior MG, anterior MG are more active in protein synthesis and produce more immunoglobulins and lactoferrin in colostrum and milk.}, number={8}, journal={Journal of Animal Science}, publisher={Oxford University Press (OUP)}, author={Wu, W. Z. and Wang, X. Q. and Wu, G. Y. and Kim, S. W. and Chen, F. and Wang, J. J.}, year={2010}, month={Aug}, pages={2657–2664} }
 @article{wang_wu_lin_li_wu_wang_2010, title={Temporal Proteomic Analysis Reveals Continuous Impairment of Intestinal Development in Neonatal Piglets with Intrauterine Growth Restriction}, volume={9}, ISSN={1535-3893 1535-3907}, url={http://dx.doi.org/10.1021/pr900747d}, DOI={10.1021/pr900747d}, abstractNote={Efficiency of nutrient utilization is reduced in neonates with intrauterine growth restriction (IUGR) compared with those with a normal birth weight (NBW). However, the underlying mechanisms are largely unknown. In this study, we applied temporal proteomic approach, coupled with histological and biochemical analyses, to study dynamic changes of the proteome in the small intestinal mucosa of IUGR piglets during the nursing period (Days 1, 7 and 21). We identified 56 differentially expressed protein spots between IUGR and NBW piglets. These proteins participate in key biological processes, including (1) absorption, digestion and transport of nutrients; (2) cell structure and motility; (3) glucose and energy metabolism; (4) lipid metabolism; (5) amino acid metabolism; (6) mineral and vitamin metabolism; (7) cellular redox homeostasis; (8) stress response; and (9) apoptosis. The results of our temporal proteomics analysis reveal continuous impairment of intestinal development in neonatal piglets with IUGR. The findings have important implications for understanding metabolic defects in the small intestine of IUGR neonates and are expected to provide new strategies to improve their survival and growth.}, number={2}, journal={Journal of Proteome Research}, publisher={American Chemical Society (ACS)}, author={Wang, Xiaoqiu and Wu, Weizong and Lin, Gang and Li, Defa and Wu, Guoyao and Wang, Junjun}, year={2010}, month={Feb}, pages={924–935} }
 @article{wu_wang_wang_2009, title={Advance in maternal bioactive compounds in promoting growth and development of piglets}, volume={45}, number={9}, journal={Chinese Journal of Animal Science}, author={Wu, W. and Wang, X. and Wang, J.}, year={2009}, pages={61–64} }
 @article{liu_han_huang_wang_wang_wang_2009, title={Dietary L-arginine Supplementation Improves Intestinal Function in Weaned Pigs after an Escherichia coli Lipopolysaccharide Challenge}, volume={22}, ISSN={1011-2367 1976-5517}, url={http://dx.doi.org/10.5713/ajas.2009.90100}, DOI={10.5713/ajas.2009.90100}, abstractNote={This study was conducted to determine whether L-arginine (Arg) supplementation could improve intestinal function in weaned pigs after an Escherichia coli lipopolysaccharide (LPS) challenge. Treatments included: i) non-challenged control (CONTR, pigs fed a control diet and injected with sterile saline); ii) LPS-challenged control (LPS, pigs fed the same control diet and challenged by injection with Escherichia coli LPS); iii) LPS+0.5% Arg (pigs fed a 0.5% Arg diet and challenged with LPS); and iv) LPS+1.0% Arg (pigs fed a 1.0% Arg diet and challenged with LPS). On d 16, pigs were administrated with LPS or sterile saline. D-xylose was orally administrated at 2 h following LPS challenge, and blood samples were collected at 3 h following LPS challenge. At 6 h post-challenge, pigs were sacrificed and intestinal mucosa samples were collected. Supplementation of Arg attenuated LPS-induced damage in gut digestive and barrier functions, as indicated by an increase in ileal lactase activity, and duodenal and ileal diamine oxidase activities (p<0.05). Arg administration also prevented the increase of jejunal malondialdehyde content and the decrease of ileal superoxide dismutase activity by LPS challenge (p<0.05). Furthermore, the jejunal nitric oxide level and inducible nitric oxide synthase activity were also improved after Arg supplementation (p<0.05). These results indicate that Arg supplementation has beneficial effects in alleviating the impairment of gut function induced by LPS challenge.}, number={12}, journal={Asian-Australasian Journal of Animal Sciences}, publisher={Asian Australasian Association of Animal Production Societies}, author={Liu, Yulan and Han, Jie and Huang, Jingjing and Wang, Xiaoqiu and Wang, Fenglai and Wang, Junjun}, year={2009}, month={Oct}, pages={1667–1675} }
 @article{wu_wang_wang_2009, title={Progress in proteome research of breast milk}, volume={21}, number={6}, journal={Chinese Journal of Animal Nutrition}, author={Wu, W. and Wang, X. and Wang, J.}, year={2009}, pages={809–815} }
 @article{wang_ou_yin_wu_wang_2009, title={Proteomic analysis reveals altered expression of proteins related to glutathione metabolism and apoptosis in the small intestine of zinc oxide-supplemented piglets}, volume={37}, ISSN={0939-4451 1438-2199}, url={http://dx.doi.org/10.1007/s00726-009-0242-y}, DOI={10.1007/s00726-009-0242-y}, number={1}, journal={Amino Acids}, publisher={Springer Science and Business Media LLC}, author={Wang, Xiaoqiu and Ou, Deyuan and Yin, Jingdong and Wu, Guoyao and Wang, Junjun}, year={2009}, month={Jan}, pages={209–218} }
 @article{lin_wang_wang_wu_lai_2009, title={The development of T lymphocytes in intrauterine growth retardation piglets}, volume={45}, number={21}, journal={Chinese Journal of Animal Science}, author={Lin, Y. and Wang, J. and Wang, X. and Wu, W. and Lai, C.}, year={2009}, pages={13–15} }
 @article{wang_chen_li_yin_wang_li_dangott_hu_wu_2008, title={Intrauterine Growth Restriction Affects the Proteomes of the Small Intestine, Liver, and Skeletal Muscle in Newborn Pigs}, volume={138}, ISSN={0022-3166 1541-6100}, url={http://dx.doi.org/10.1093/jn/138.1.60}, DOI={10.1093/jn/138.1.60}, abstractNote={Efficiency of nutrient utilization is high in neonates with normal birth weights but is reduced in those with intrauterine growth restriction (IUGR). However, the underlying mechanisms are largely unknown. This study was conducted with the piglet model and proteomics technology to test the hypothesis that IUGR affects expression of key proteins that regulate growth and development of the small intestine, liver, and muscle, the major organs involved in the digestion, absorption, and metabolism of dietary nutrients. Jejunum, liver, and gastrocnemius muscle were obtained from IUGR and normal birth-weight piglets at birth for analysis of proteomes using the 2-dimensional-PAGE MS technology. The results indicate that IUGR decreased the levels of proteins that regulate immune function (immunoglobulins and annexin A1), oxidative defense (peroxiredoxin 1, transferrin, and zeta-crystallin), intermediary metabolism (creatine kinase, alcohol dehydrogenase, L-lactate dehydrogenase, prostaglandin F synthase, apolipoprotein AI, catecho O-methyltransferase, and phosphoglycerate kinase 1), protein synthesis (eukaryotic translation initiation factor-3), and tissue growth (beta-actin, desmin, and keratin 10) in a tissue-specific manner. In addition, IUGR increased the levels of proteins that are involved in proteolysis (proteasome alpha-5 and alpha-1 subunits), response to oxidative stress (scavenger-receptor protein and alpha-1 acid glycoprotein), and ATP hydrolysis (F1-ATPase). These novel findings suggest that cellular signaling defects, redox imbalance, reduced protein synthesis, and enhanced proteolysis may be the major mechanisms responsible for abnormal absorption and metabolism of nutrients, as well as reduced growth and impaired development of the small intestine, liver, and muscle in IUGR neonates.}, number={1}, journal={The Journal of Nutrition}, publisher={Oxford University Press (OUP)}, author={Wang, Junjun and Chen, Lixiang and Li, Defa and Yin, Yulong and Wang, Xiaoqiu and Li, Peng and Dangott, Lawrence J. and Hu, Weixin and Wu, Guoyao}, year={2008}, month={Jan}, pages={60–66} }