@article{grzymkowski_chiu_jima_wyatt_jayachandran_stutts_nascone-yoder_2024, title={Developmental regulation of cellular metabolism is required for intestinal elongation and rotation}, volume={151}, ISSN={["1477-9129"]}, url={https://doi.org/10.1242/dev.202020}, DOI={10.1242/dev.202020}, abstractNote={ABSTRACT Malrotation of the intestine is a prevalent birth anomaly, the etiology of which remains poorly understood. Here, we show that late-stage exposure of Xenopus embryos to atrazine, a widely used herbicide that targets electron transport chain (ETC) reactions, elicits intestinal malrotation at high frequency. Interestingly, atrazine specifically inhibits the cellular morphogenetic events required for gut tube elongation, including cell rearrangement, differentiation and proliferation; insufficient gut lengthening consequently reorients the direction of intestine rotation. Transcriptome analyses of atrazine-exposed intestines reveal misexpression of genes associated with glycolysis and oxidative stress, and metabolomics shows that atrazine depletes key glycolytic and tricarboxylic acid cycle metabolites. Moreover, cellular bioenergetics assays indicate that atrazine blocks a crucial developmental transition from glycolytic ATP production toward oxidative phosphorylation. Atrazine-induced defects are phenocopied by rotenone, a known ETC Complex I inhibitor, accompanied by elevated reactive oxygen species, and rescued by antioxidant supplementation, suggesting that malrotation may be at least partly attributable to redox imbalance. These studies reveal roles for metabolism in gut morphogenesis and implicate defective gut tube elongation and/or metabolic perturbations in the etiology of intestinal malrotation.}, number={1}, journal={DEVELOPMENT}, author={Grzymkowski, Julia K. and Chiu, Yu-Chun and Jima, Dereje D. and Wyatt, Brent H. and Jayachandran, Sudhish and Stutts, Whitney L. and Nascone-Yoder, Nanette M.}, year={2024}, month={Jan} } @article{barnes_rodriguez-zapata_juarez-nunez_gates_janzen_kur_wang_jensen_estevez-palmas_crow_et al._2022, title={An adaptive teosinte mexicana introgression modulates phosphatidylcholine levels and is associated with maize flowering time}, volume={119}, ISSN={["1091-6490"]}, url={http://dx.doi.org/10.1073/pnas.2100036119}, DOI={10.1073/pnas.2100036119}, abstractNote={Native Americans domesticated maize ( Zea mays ssp. mays ) from lowland teosinte parviglumis ( Zea mays ssp. parviglumis) in the warm Mexican southwest and brought it to the highlands of Mexico and South America where it was exposed to lower temperatures that imposed strong selection on flowering time. Phospholipids are important metabolites in plant responses to low-temperature and phosphorus availability and have been suggested to influence flowering time. Here, we combined linkage mapping with genome scans to identify High PhosphatidylCholine 1 ( HPC1 ), a gene that encodes a phospholipase A1 enzyme, as a major driver of phospholipid variation in highland maize. Common garden experiments demonstrated strong genotype-by-environment interactions associated with variation at HPC1, with the highland HPC1 allele leading to higher fitness in highlands, possibly by hastening flowering. The highland maize HPC1 variant resulted in impaired function of the encoded protein due to a polymorphism in a highly conserved sequence. A meta-analysis across HPC1 orthologs indicated a strong association between the identity of the amino acid at this position and optimal growth in prokaryotes. Mutagenesis of HPC1 via genome editing validated its role in regulating phospholipid metabolism. Finally, we showed that the highland HPC1 allele entered cultivated maize by introgression from the wild highland teosinte Zea mays ssp. mexicana and has been maintained in maize breeding lines from the Northern United States, Canada, and Europe. Thus, HPC1 introgressed from teosinte mexicana underlies a large metabolic QTL that modulates phosphatidylcholine levels and has an adaptive effect at least in part via induction of early flowering time.}, number={27}, journal={PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA}, publisher={Proceedings of the National Academy of Sciences}, author={Barnes, Allison C. and Rodriguez-Zapata, Fausto and Juarez-Nunez, Karla A. and Gates, Daniel J. and Janzen, Garrett M. and Kur, Andi and Wang, Li and Jensen, Sarah E. and Estevez-Palmas, Juan M. and Crow, Taylor M. and et al.}, year={2022}, month={Jul} } @book{barnes_rodrı́guez-zapata fausto_blöcher-juárez_gates_kur_wang_janzen_jensen_estévez-palmas_crow_et al._2021, title={Teosinte introgression modulates phosphatidylcholine levels and induces early maize flowering time}, volume={1}, url={http://dx.doi.org/10.1101/2021.01.25.426574}, DOI={10.1101/2021.01.25.426574}, abstractNote={After domestication from lowland teosinte parviglumis (Zea mays ssp.parviglumis) in the warm Mexican southwest, maize (Zea mays ssp. mays) colonized the highlands of México and South America where it was exposed to lower temperatures that imposed strong selection on flowering time. Phospholipids are important metabolites in plant responses to low-temperature and low phosphorus availability, and have also been suggested to influence flowering time. Here, we combined linkage mapping with genome scans to identify High PhosphatidylCholine 1 (HPC1), a gene that encodes a phospholipase A1 enzyme, as a major driver of phospholipid variation in highland maize. Common garden experiments demonstrated strong genotype-by-environment interactions associated with variation at HPC1, with the highland HPC1 allele leading to higher fitness in highlands, possibly by hastening flowering. The highland maize HPC1 variant resulted in impaired function of the encoded protein due to a polymorphism in a highly conserved sequence. A meta-analysis across HPC1 orthologs indicated a strong association between the identity of the amino acid at this position and optimal growth in prokaryotes. Mutagenesis of HPC1 via genome editing validated its role in regulating phospholipid metabolism. Finally, we showed that the highland HPC1 allele entered cultivated maize by introgression from the wild highland teosinte Zea mays ssp. mexicana and has been maintained in maize breeding lines from the Northern US, Canada and Europe. Thus, HPC1 introgressed from teosinte mexicana underlies a large metabolic QTL that modulates phosphatidylcholine levels and has an adaptive effect at least in part via induction of early flowering time. Significance Statement Despite more than a century of genetic research, our understanding of the genetic basis of the astounding capacity of maize to adapt to new environments is in its infancy. Recent work in many crops has pointed to the potentially important role for introgression in underpinning adaptation, but clear examples of adaptive loci arising via introgression are lacking. Here, we elucidate the evolutionary history of a novel, major metabolic quantitative trait locus (QTL) that we mapped down to a single gene, HPC1. Alterations in highland HPC1 are the result of a teosinte mexicana introgression in maize, leading to high phosphatidylcholine levels and improving fitness by accelerating flowering.}, journal={[]}, institution={Cold Spring Harbor Laboratory}, author={Barnes, Allison C and Rodrı́guez-Zapata Fausto and Blöcher-Juárez, Karla A and Gates, Dan J and Kur, Andi and Wang, Li and Janzen, Garrett M and Jensen, Sarah E and Estévez-Palmas, Juan M and Crow, Taylor M and et al.}, year={2021}, month={Jan} } @article{knuth_stutts_ritter_garrard_kullman_2021, title={Vitamin D deficiency promotes accumulation of bioactive lipids and increased endocannabinoid tone in zebrafish}, volume={62}, ISSN={["1539-7262"]}, url={https://doi.org/10.1016/j.jlr.2021.100142}, DOI={10.1016/j.jlr.2021.100142}, abstractNote={Vitamin D is well known for its traditional role in bone mineral homeostasis; however, recent evidence suggests that vitamin D also plays a significant role in metabolic control. This study served to investigate putative linkages between vitamin D deficiency (VDD) and metabolic disruption of bioactive lipids by MS imaging. Our approach employed infrared-matrix-assisted laser desorption electrospray ionization MS imaging for lipid metabolite profiling in 6-month-old zebrafish fed either a VDD or a vitamin D-sufficient (VDS) diet. Using a lipidomics pipeline, we found that VDD zebrafish had a greater abundance of bioactive lipids (N-acyls, endocannabinoids [ECs], diacylglycerols/triacylglycerols, bile acids/bile alcohols, and vitamin D derivatives) suggestive of increased EC tone compared with VDS zebrafish. Tandem MS was performed on several differentially expressed metabolites with sufficient ion abundances to aid in structural elucidation and provide additional support for MS annotations. To confirm activation of the EC pathways, we subsequently examined expression of genes involved in EC biosynthesis, metabolism, and receptor signaling in adipose tissue and liver from VDD and VDS zebrafish. Gene expression changes were congruent with increased EC tone, with VDD zebrafish demonstrating increased synthesis and metabolism of anandamide compared with VDS zebrafish. Taken together, our data suggest that VDD may promote accumulation of bioactive lipids and increased EC tone in zebrafish.}, journal={JOURNAL OF LIPID RESEARCH}, publisher={Elsevier BV}, author={Knuth, Megan M. and Stutts, Whitney L. and Ritter, Morgan M. and Garrard, Kenneth P. and Kullman, Seth W.}, year={2021} } @article{wolny_egerton_handy_stutts_smith_whereat_bachvaroff_henrichs_campbell_deeds_2020, title={Characterization of Dinophysis spp. (Dinophyceae, Dinophysiales) from the mid-Atlantic region of the United States}, volume={56}, ISSN={["1529-8817"]}, url={http://dx.doi.org/10.1111/jpy.12966}, DOI={10.1111/jpy.12966}, abstractNote={Due to the increasing prevalence of Dinophysis spp. and their toxins on every US coast in recent years, the need to identify and monitor for problematic Dinophysis populations has become apparent. Here, we present morphological analyses, using light and scanning electron microscopy, and rDNA sequence analysis, using a ~2‐kb sequence of ribosomal ITS1, 5.8S, ITS2, and LSU DNA, of Dinophysis collected in mid‐Atlantic estuarine and coastal waters from Virginia to New Jersey to better characterize local populations. In addition, we analyzed for diarrhetic shellfish poisoning (DSP) toxins in water and shellfish samples collected during blooms using liquid‐chromatography tandem mass spectrometry and an in vitro protein phosphatase inhibition assay and compared this data to a toxin profile generated from a mid‐Atlantic Dinophysis culture. Three distinct morphospecies were documented in mid‐Atlantic surface waters: D. acuminata, D. norvegica, and a “small Dinophysis sp.” that was morphologically distinct based on multivariate analysis of morphometric data but was genetically consistent with D. acuminata. While mid‐Atlantic D. acuminata could not be distinguished from the other species in the D. acuminata‐complex (D. ovum from the Gulf of Mexico and D. sacculus from the western Mediterranean Sea) using the molecular markers chosen, it could be distinguished based on morphometrics. Okadaic acid, dinophysistoxin 1, and pectenotoxin 2 were found in filtered water and shellfish samples during Dinophysis blooms in the mid‐Atlantic region, as well as in a locally isolated D. acuminata culture. However, DSP toxins exceeded regulatory guidance concentrations only a few times during the study period and only in noncommercial shellfish samples.}, number={2}, journal={JOURNAL OF PHYCOLOGY}, author={Wolny, Jennifer L. and Egerton, Todd A. and Handy, Sara M. and Stutts, Whitney L. and Smith, Juliette L. and Whereat, Edward B. and Bachvaroff, Tsvetan R. and Henrichs, Darren W. and Campbell, Lisa and Deeds, Jonathan R.}, year={2020}, month={Apr}, pages={404–424} } @article{dihydrodinophysistoxin-1 produced by dinophysis norvegica in the gulf of maine, usa and its accumulation in shellfish._2020, url={https://europepmc.org/articles/PMC7551465}, DOI={10.3390/toxins12090533}, abstractNote={Dihydrodinophysistoxin-1 (dihydro-DTX1, (M-H)− m/z 819.5), described previously from a marine sponge but never identified as to its biological source or described in shellfish, was detected in multiple species of commercial shellfish collected from the central coast of the Gulf of Maine, USA in 2016 and in 2018 during blooms of the dinoflagellate Dinophysis norvegica. Toxin screening by protein phosphatase inhibition (PPIA) first detected the presence of diarrhetic shellfish poisoning-like bioactivity; however, confirmatory analysis using liquid chromatography-tandem mass spectrometry (LC-MS/MS) failed to detect okadaic acid (OA, (M-H)− m/z 803.5), dinophysistoxin-1 (DTX1, (M-H)− m/z 817.5), or dinophysistoxin-2 (DTX2, (M-H)− m/z 803.5) in samples collected during the bloom. Bioactivity-guided fractionation followed by liquid chromatography-high resolution mass spectrometry (LC-HRMS) tentatively identified dihydro-DTX1 in the PPIA active fraction. LC-MS/MS measurements showed an absence of OA, DTX1, and DTX2, but confirmed the presence of dihydro-DTX1 in shellfish during blooms of D. norvegica in both years, with results correlating well with PPIA testing. Two laboratory cultures of D. norvegica isolated from the 2018 bloom were found to produce dihydro-DTX1 as the sole DSP toxin, confirming the source of this compound in shellfish. Estimated concentrations of dihydro-DTX1 were >0.16 ppm in multiple shellfish species (max. 1.1 ppm) during the blooms in 2016 and 2018. Assuming an equivalent potency and molar response to DTX1, the authority initiated precautionary shellfish harvesting closures in both years. To date, no illnesses have been associated with the presence of dihydro-DTX1 in shellfish in the Gulf of Maine region and studies are underway to determine the potency of this new toxin relative to the currently regulated DSP toxins in order to develop appropriate management guidance.}, journal={Toxins}, year={2020}, month={Aug} } @article{stutts_knuth_ekelöf_mahapatra_kullman_muddiman_2020, title={Methods for Cryosectioning and Mass Spectrometry Imaging of Whole-Body Zebrafish}, volume={31}, ISSN={1044-0305 1879-1123}, url={http://dx.doi.org/10.1021/jasms.9b00097}, DOI={10.1021/jasms.9b00097}, abstractNote={The zebrafish (Danio rerio) is an ideal model for whole animal studies of lipid metabolism and lipid-related disease. In this work, infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) mass spectrometry imaging (MSI) was applied for direct visualization of lipid and metabolite distributions across various organs in whole-body zebrafish tissue sections. Detailed methods for overcoming the challenges of cryosectioning adult male zebrafish for MSI and complementary histological imaging are described. Representative two-dimensional ion maps demonstrated organ specific localization of lipid analytes allowing for visualization of areas of interest including the brain, liver, intestines, and skeletal muscle. A high resolving power mass spectrometer was utilized for accurate mass measurements, which permitted the use of open-source, web-based tools for MS1 annotations including METASPACE and METLIN. Whole-body MSI with IR-MALDESI allowed for broad lipid coverage with high spatial resolution, illustrating the potential of this technique for studying lipid-related diseases using zebrafish as a model organism.}, number={4}, journal={Journal of the American Society for Mass Spectrometry}, publisher={American Chemical Society (ACS)}, author={Stutts, Whitney L. and Knuth, Megan M. and Ekelöf, Måns and Mahapatra, Debabrata and Kullman, Seth W. and Muddiman, David C.}, year={2020}, month={Feb}, pages={768–772} } @article{microcystin toxins at potentially hazardous levels in algal dietary supplements revealed by a combination of bioassay, immunoassay, and mass spectrometric methods._2020, url={https://doi.org/10.1021/acs.jafc.0c02024}, DOI={10.1021/acs.jafc.0c02024}, abstractNote={Microcystins (MCs) are hepatotoxic heptapeptides produced by cyanobacteria and are potent inhibitors of protein phosphatases in eukaryotic cells. Algae for dietary supplements are harvested from outdoor environments and can be contaminated with MCs. Monitoring of MCs in these products is necessary, but is complicated by their structural diversity (> 250 congeners). We used a combination of protein phosphatase inhibition assay (PPIA), ELISA, LC-MS/MS and nontargeted LC-high resolution MS (LC-HRMS) with thiol derivatization to characterize the total MCs in eighteen algal dietary supplements. LC-MS/MS revealed some products contained >40 times the maximum acceptable concentration (MAC) of 1 μg/g MCs, but ELISA and PPIA showed up to 50-60 times the MAC. LC-HRMS identified all congeners targeted by LC-MS/MS plus MC-(H4)YR contributing up to 18% of total MCs, along with numerous minor MCs. Recommended dosages of the products with >MAC would result in 2.6-75 times the tolerable daily intake, presenting a risk to consumers. This study confirms the need for monitoring these products, and presents strategies to fully describe the total MC pool in environmental samples and algal products.}, journal={Journal of agricultural and food chemistry}, year={2020}, month={Jul} } @article{mazzola_deeds_stutts_ridge_dickey_white_williamson_martin_2019, title={Elucidation and partial NMR assignment of monosulfated maitotoxins from the Caribbean}, volume={164}, ISSN={["0041-0101"]}, url={http://www.scopus.com/inward/record.url?eid=2-s2.0-85064154618&partnerID=MN8TOARS}, DOI={10.1016/j.toxicon.2019.03.026}, abstractNote={Compounds similar to maitotoxin (MTX) have been isolated from several laboratory strains of the dinoflagellate Gambierdiscus spp. from the Caribbean. Mass spectral results suggest that these compounds differ from MTX by the loss of one sulfate group and, in some cases, the loss of one methyl group with the addition of one degree of unsaturation. NMR experiments, using approximately 50 nmol of one of these compounds, have demonstrated that the 9-sulfo group of MTX is still present, suggesting that these compounds are 40-desulfo congeners of MTX.}, journal={TOXICON}, author={Mazzola, Eugene P. and Deeds, Jonathan R. and Stutts, Whitney L. and Ridge, Clark D. and Dickey, Robert W. and White, Kevin D. and Williamson, R. Thomas and Martin, Gary E.}, year={2019}, month={Jun}, pages={44–50} } @article{marsan_conrad_stutts_parker_deeds_2018, title={Evaluation of microcystin contamination in blue-green algal dietary supplements using a protein phosphatase inhibition-based test kit}, volume={4}, url={http://www.scopus.com/inward/record.url?eid=2-s2.0-85043994824&partnerID=MN8TOARS}, DOI={10.1016/j.heliyon.2018.e00573}, abstractNote={