@article{kouprianov_selmek_ferguson_mo_shive_2022, title={brca2-mutant zebrafish exhibit context- and tissue-dependent alterations in cell phenotypes and response to injury}, volume={12}, ISSN={["2045-2322"]}, DOI={10.1038/s41598-022-04878-9}, abstractNote={AbstractCancer cells frequently co-opt molecular programs that are normally activated in specific contexts, such as embryonic development and the response to injury. Determining the impact of cancer-associated mutations on cellular phenotypes within these discrete contexts can provide new insight into how such mutations lead to dysregulated cell behaviors and subsequent cancer onset. Here we assess the impact of heritable BRCA2 mutation on embryonic development and the injury response using a zebrafish model (Danio rerio). Unlike most mouse models for BRCA2 mutation, brca2-mutant zebrafish are fully viable and thus provide a unique tool for assessing both embryonic and adult phenotypes. We find that maternally provided brca2 is critical for normal oocyte development and embryonic survival in zebrafish, suggesting that embryonic lethality associated with BRCA2 mutation is likely to reflect defects in both meiotic and embryonic developmental programs. On the other hand, we find that adult brca2-mutant zebrafish exhibit aberrant proliferation of several cell types under basal conditions and in response to injury in tissues at high risk for cancer development. These divergent effects exemplify the often-paradoxical outcomes that occur in embryos (embryonic lethality) versus adult animals (cancer predisposition) with mutations in cancer susceptibility genes such as BRCA2. The altered cell behaviors identified in brca2-mutant embryonic and adult tissues, particularly in adult tissues at high risk for cancer, indicate that the effects of BRCA2 mutation on cellular phenotypes are both context- and tissue-dependent.}, number={1}, journal={SCIENTIFIC REPORTS}, author={Kouprianov, Vassili A. and Selmek, Aubrie A. and Ferguson, Jordan L. and Mo, Xiaokui and Shive, Heather R.}, year={2022}, month={Jan} }
 @article{womble_lewbart_shive_2020, title={Pathologic Lesions of the Budgett Frog (Lepidobatrachus laevis), an Emerging Laboratory Animal Model}, volume={70}, ISSN={["1532-0820"]}, DOI={10.30802/AALAS-CM-19-000071}, abstractNote={Lepidobatrachus laevis, commonly called the Budgett frog, is a member of the horned frog family (Ceratophryidae), which has become increasingly popular among amphibian hobbyists. L. laevis is also used in biologic research on embryonic development, providing a novel model
 species for the study of organogenesis, regeneration, evolution, and biologic scaling. However, little scientific literature details disease processes or histologic lesions in this species. Our objective was to describe spontaneous pathologic lesions in L. laevis to identify disease
 phenotypes. We performed a retrospective analysis of 14 captive L. laevis frogs (wild-caught and captive-bred), necropsied at the NC State University College of Veterinary Medicine between 2008 and 2018. The majority of frogs exhibited renal changes, including varying combinations of
 tubular epithelial binucleation, karyomegaly, and cytoplasmic vacuolation; polycystic kidney disease; and renal carcinoma. Many of the renal changes are reminiscent of a condition described in Japanese (Bufo japonicus) and Chinese (Bufo raddei) toad hybrids that progresses from
 tubular epithelial atypia and tubular dilation to polycystic kidney disease to renal carcinoma. A second common finding was variably sized, randomly distributed bile duct clusters (biliary proliferation). Other noteworthy findings included regional or generalized edema, intestinal adenocarcinoma,
 aspiration pneumonia, and parasitism. This retrospective analysis is the first description of histologic lesions identified in captive L. laevis populations, providing new insight into spontaneous disease processes occurring in this species for use in disease diagnosis and clinical
 management.}, number={3}, journal={COMPARATIVE MEDICINE}, author={Womble, Mandy A. and Lewbart, Gregory A. and Shive, Heather R.}, year={2020}, month={Jun}, pages={239–247} }
 @article{mensah_ferguson_shive_2019, title={Genotypic and Phenotypic Variables Affect Meiotic Cell Cycle Progression, Tumor Ploidy, and Cancer-Associated Mortality in a brca2-Mutant Zebrafish Model}, volume={2019}, ISSN={["1687-8469"]}, DOI={10.1155/2019/9218251}, abstractNote={Successful cell replication requires both cell cycle completion and accurate chromosomal segregation. The tumor suppressor BRCA2 is positioned to influence both of these outcomes, and thereby influence genomic integrity, during meiotic and mitotic cell cycles. Accordingly, mutations in BRCA2 induce chromosomal abnormalities and disrupt cell cycle progression in both germ cells and somatic cells. Despite these findings, aneuploidy is not more prevalent in BRCA2-associated versus non-BRCA2-associated human cancers. More puzzlingly, diploidy in BRCA2-associated cancers is a negative prognostic factor, unlike non-BRCA2-associated cancers and many other human cancers. We used a brca2-mutant/tp53-mutant cancer-prone zebrafish model to explore the impact of BRCA2 mutation on cell cycle progression, ploidy, and cancer-associated mortality by performing DNA content/cell cycle analysis on zebrafish germ cells, somatic cells, and cancer cells. First, we determined that combined brca2/tp53 mutations uniquely disrupt meiotic progression. Second, we determined that sex significantly influences ploidy outcome in zebrafish cancers. Third, we determined that brca2 mutation and female sex each significantly reduce survival time in cancer-bearing zebrafish. Finally, we provide evidence to support a link between BRCA2 mutation, tumor diploidy, and poor survival outcome. These outcomes underscore the utility of this model for studying BRCA2-associated genomic aberrations in normal and cancer cells.}, journal={JOURNAL OF ONCOLOGY}, author={Mensah, L. and Ferguson, J. L. and Shive, H. R.}, year={2019} }
 @article{ferguson_shive_2019, title={Sequential Immunofluorescence and Immunohistochemistry on Cryosectioned Zebrafish Embryos}, ISSN={["1940-087X"]}, DOI={10.3791/59344}, abstractNote={Investigation of intercellular interactions often requires discrete labeling of specific cell populations and precise protein localization. The zebrafish embryo is an excellent tool for examining such interactions with an in vivo model. Whole-mount immunohistochemical and immunofluorescence assays are frequently applied in zebrafish embryos to assess protein expression. However, it can be difficult to achieve accurate mapping of co-localized proteins in three-dimensional space. In addition, some studies may require the use of two antibodies that are not compatible with the same technique (e.g., antibody 1 is only suitable for immunohistochemistry and antibody 2 is only suitable for immunofluorescence). The purpose of the method described herein is to perform sequential immunofluorescence and/or immunohistochemistry on individual cryosections derived from early-stage zebrafish embryos. Here we describe the use of sequential rounds of immunofluorescence, imaging, immunohistochemistry, imaging for a single cryosection in order to achieve precise identification of protein expression at the single-cell level. This methodology is suitable for any study in early-stage zebrafish embryos that requires accurate identification of multiple protein targets in individual cells.}, number={147}, journal={JOVE-JOURNAL OF VISUALIZED EXPERIMENTS}, author={Ferguson, Jordan L. and Shive, Heather R.}, year={2019}, month={May} }
 @article{schultz_haltom_almeida_wierson_solin_weiss_helmer_sandquist_shive_mcgrail_2018, title={Epigenetic regulators Rbbp4 and Hdac1 are overexpressed in a zebrafish model of RB1 embryonal brain tumor, and are required for neural progenitor survival and proliferation}, volume={11}, ISSN={["1754-8411"]}, DOI={10.1242/dmm.034124}, abstractNote={ABSTRACT
               In this study, we used comparative genomics and developmental genetics to identify epigenetic regulators driving oncogenesis in a zebrafish retinoblastoma 1 (rb1) somatic-targeting model of RB1 mutant embryonal brain tumors. Zebrafish rb1 brain tumors caused by TALEN or CRISPR targeting are histologically similar to human central nervous system primitive neuroectodermal tumors (CNS-PNETs). Like the human oligoneural OLIG2+/SOX10+ CNS-PNET subtype, zebrafish rb1 tumors show elevated expression of neural progenitor transcription factors olig2, sox10, sox8b and the receptor tyrosine kinase erbb3a oncogene. Comparison of rb1 tumor and rb1/rb1 germline mutant larval transcriptomes shows that the altered oligoneural precursor signature is specific to tumor tissue. More than 170 chromatin regulators were differentially expressed in rb1 tumors, including overexpression of chromatin remodeler components histone deacetylase 1 (hdac1) and retinoblastoma binding protein 4 (rbbp4). Germline mutant analysis confirms that zebrafish rb1, rbbp4 and hdac1 are required during brain development. rb1 is necessary for neural precursor cell cycle exit and terminal differentiation, rbbp4 is required for survival of postmitotic precursors, and hdac1 maintains proliferation of the neural stem cell/progenitor pool. We present an in vivo assay using somatic CRISPR targeting plus live imaging of histone-H2A.F/Z-GFP fusion protein in developing larval brain to rapidly test the role of chromatin remodelers in neural stem and progenitor cells. Our somatic assay recapitulates germline mutant phenotypes and reveals a dynamic view of their roles in neural cell populations. Our study provides new insight into the epigenetic processes that might drive pathogenesis in RB1 brain tumors, and identifies Rbbp4 and its associated chromatin remodeling complexes as potential target pathways to induce apoptosis in RB1 mutant brain cancer cells.
               This article has an associated First Person interview with the first author of the paper.}, number={6}, journal={DISEASE MODELS & MECHANISMS}, author={Schultz, Laura E. and Haltom, Jeffrey A. and Almeida, Maira P. and Wierson, Wesley A. and Solin, Staci L. and Weiss, Trevor J. and Helmer, Jordan A. and Sandquist, Elizabeth J. and Shive, Heather R. and McGrail, Maura}, year={2018}, month={Jun} }
 @misc{cohen_shive_borst_almeida_2018, title={Lameness in a 3-year-old backyard chicken Response}, volume={252}, number={6}, journal={Journal of the American Veterinary Medical Association}, author={Cohen, E. B. and Shive, H. R. and Borst, L. B. and Almeida, S. M. B.}, year={2018}, pages={648–648} }
 @article{roode_shive_hoorntje_bernard_stowe_pool_grindem_2018, title={Multiloculated solitary (unicameral) bone cyst in a young dog}, volume={47}, ISSN={0275-6382}, url={http://dx.doi.org/10.1111/vcp.12618}, DOI={10.1111/vcp.12618}, abstractNote={AbstractA 20‐month‐old female spayed Staffordshire Terrier (22.3 kg) presented to the Orthopedic Surgery Service at North Carolina State University Veterinary Teaching Hospital for evaluation of a 6‐week history of toe‐touching to nonweight‐bearing lameness in the right hind limb. Radiographs of the right stifle revealed a multiloculated lytic lesion of the distal femur, with a large open lytic zone centrally, numerous osseous septations peripherally, and focal areas of cortical thinning and loss. An aspirate of the right distal femoral lesion yielded mildly cloudy serosanguineous fluid. Cytologic examination of the fluid revealed a pleomorphic population of discrete cells that exhibited marked anisocytosis and anisokaryosis and a variable nuclear‐to‐cytoplasmic (N:C) ratio, which were interpreted as probable neoplastic cells, with few macrophages, and evidence of hemorrhage. Given the clinical signs of pain, lesion size, and concern for malignant neoplasia, amputation of the right hind limb was performed. Histologically, the lesion had undulating walls 1‐3 mm thick with a continuous outer layer of dense fibrous tissue and an inner layer composed of reactive cancellous bone with no cortical compacta remaining. Remnants of thin fibrous or fibro‐osseous septa projected from the bony wall into the cyst lumen. The final histologic diagnosis was a benign multiloculated solitary (unicameral) bone cyst of the distal right femur. Based on the histopathologic findings, it was speculated that the cells identified on cytology were a mixture of developing osteoclasts, osteoblasts, endothelial, and stromal cells. This is the first report describing the cytologic examination of a solitary bone cyst in veterinary medicine.}, number={3}, journal={Veterinary Clinical Pathology}, publisher={Wiley}, author={Roode, Sarah C. and Shive, Heather R. and Hoorntje, Willemijn and Bernard, Jennifer and Stowe, Devorah M. and Pool, Roy R. and Grindem, Carol B.}, year={2018}, month={May}, pages={484–488} }
 @article{almeida_shive_harvey_borst_cohen_2018, title={What is your diagnosis?}, volume={252}, url={http://www.scopus.com/inward/record.url?eid=2-s2.0-85040519711&partnerID=MN8TOARS}, DOI={10.2460/javma.252.2.173}, abstractNote={On clinical examination, the dog was bright, alert and responsive with a body weight of 17.2kg and a body condition score of 5/9. The mucous membranes were pink and moist, with a capillary refill time of one second. The heart rate was 140 beats/min but the dog was quite nervous. The dog was panting but no adventitious lung sounds were auscultated. A clear abdominal fluid thrill was detected but there were no other specific finding on palpation. The rectal temperature was normal at 38.9°C.}, number={2}, journal={Journal of the American Veterinary Medical Association}, author={Almeida, S.M. Bessauer and Shive, H.R. and Harvey, J.B. and Borst, Luke and Cohen, Eli}, year={2018}, pages={173–175} }
 @article{williams_long_durrant_mckeon_shive_griffith_messenger_fish_2017, title={Oral transmucosal detomidine gel in New Zealand white rabbits (Oryctolagus cuniculus)}, volume={56}, number={4}, journal={Journal of the American Association for Laboratory Animal Science}, author={Williams, M. D. and Long, C. T. and Durrant, J. R. and McKeon, G. P. and Shive, H. R. and Griffith, E. H. and Messenger, K. M. and Fish, R. E.}, year={2017}, pages={436–442} }
 @article{white_sexton_shive_2017, title={Histologic and immunohistochemical analyses of soft tissue sarcomas from brca2-mutant/tp53-mutant zebrafish are consistent with neural crest (Schwann Cell) origin}, volume={54}, DOI={10.1177/0300985816669406}, abstractNote={The zebrafish ( Danio rerio) provides a powerful model for analyzing genetic contributors to cancer. Multiple zebrafish lines with cancer-associated genetic mutations develop soft tissue sarcomas that are histologically consistent with malignant peripheral nerve sheath tumor (MPNST). The goal of this study was to determine the phenotype of soft tissue sarcomas in a brca2-mutant/ tp53-mutant zebrafish line using immunohistochemical markers that are commonly expressed in mammalian MPNST. We classified 70 soft tissue sarcomas from a brca2-mutant/ tp53-mutant zebrafish cohort as MPNST, undifferentiated sarcoma, or other tumor based on histologic features. The expression of S100, CD57, and glial fibrillary acidic protein (GFAP) was analyzed in nonneoplastic neural tissues and tumor specimens by immunohistochemistry. Each marker was expressed in nonneoplastic neural tissues. In MPNST, S100 and CD57 were widely expressed in neoplastic cells, with greater consistency observed for CD57 expression. In undifferentiated sarcomas, results were variable and correlated to anatomic location. Coelomic undifferentiated sarcomas often exhibited widespread CD57 expression but limited S100 expression. In comparison, ocular undifferentiated sarcomas exhibited limited expression of both CD57 and S100. Overall, CD57 and S100 expression was significantly higher in MPNST than in undifferentiated sarcomas. GFAP was not expressed in any of the tumors. This study identified commercially available antibodies that are useful for analyzing S100, CD57, and GFAP expression in zebrafish. This study further shows a correlation between degree of histologic differentiation and expression of these markers in soft tissue sarcomas from brca2-mutant/ tp53-mutant zebrafish and suggests that these cancers are derived from the neural crest with differentiation toward myelinating Schwann cells.}, number={2}, journal={Veterinary Pathology}, author={White, L. A. and Sexton, J. M. and Shive, H. R.}, year={2017}, pages={320–327} }
 @article{shive_west_embree_sexton_hickstein_2015, title={Expression of KRAS(G12V) in Zebrafish Gills Induces Hyperplasia and CXCL8-Associated Inflammation}, volume={12}, ISSN={["1557-8542"]}, DOI={10.1089/zeb.2014.1038}, abstractNote={Abstract The zebrafish (Danio rerio) represents an important animal model for analyzing genetic contributors to carcinogenesis. To assess the role for mutationally activated Ras in ovarian cancer, we developed a transgenic zebrafish model using the putative promoter for zebrafish insulin-like growth factor 3 (igf3) to drive expression of the human oncogene KRASG12V fused to EGFP. A member of the IGF family, igf3 is unique to teleosts and reportedly exhibits gonad-specific expression in fish species. In contrast to previous studies, we observed igf3 expression in wild-type zebrafish gills in addition to gonads, indicating that igf3 expression is not necessarily gonad specific. In transgenic zebrafish, expression of EGFP-KRASG12V driven by the igf3 promoter occurred only in the gills and resulted in proliferation of a putative progenitor cell population, chondroid hyperplasia, and localized inflammation. KRASG12V-transformed cells in transgenic zebrafish showed activation of the ERK-MAP kinase pathway and e...}, number={3}, journal={ZEBRAFISH}, author={Shive, Heather R. and West, Robert R. and Embree, Lisa J. and Sexton, Jamie M. and Hickstein, Dennis D.}, year={2015}, month={Jun}, pages={221–229} }
 @article{solin_shive_woolard_essner_mcgrail_2015, title={Rapid tumor induction in zebrafish by TALEN-mediated somatic inactivation of the retinoblastoma1 tumor suppressor rb1}, volume={5}, ISSN={["2045-2322"]}, DOI={10.1038/srep13745}, abstractNote={AbstractInvestigating the in vivo role of tumor suppressor genes in cancer is technically challenging due to their essential requirement during early animal development. To address this bottleneck, we generated genetic mosaic adult zebrafish using TALEN genome editing and demonstrate somatic inactivation of the tumor suppressor retinoblastoma1 (rb1) induces tumorigenesis at high frequency. 11–33% of 1-cell stage embryos injected with TALEN mRNAs targeting rb1 exon 2 or 3 develop tumors beginning as early as 3.5 months of age. Lesions predominantly arise in the brain and show features of neuroectodermal-like and glial-like tumors. Mutant allele analysis is consistent with tumor initiation due to somatic inactivation of rb1, revealing a conserved role for rb1 in tumor suppression across vertebrates. In contrast to genetic mosaics, heterozygous rb1−/+ adults show no evidence of neoplasia, while homozygous mutant rb1−/− are larval lethal. This is the first demonstration that somatic inactivation of a tumor suppressor causes cancer in zebrafish and highlights the utility of site-specific nucleases to create genetic mosaic zebrafish for tumor suppressor gene discovery. Somatic inactivation with site-directed nucleases in zebrafish presents a rapid and scalable strategy to study tumor suppressor gene function in cancer.}, journal={SCIENTIFIC REPORTS}, author={Solin, Staci L. and Shive, Heather R. and Woolard, Kevin D. and Essner, Jeffrey J. and McGrail, Maura}, year={2015}, month={Sep} }
 @article{bauer_tuschong_calvo_shive_burkholder_karlsson_west_russell_hickstein_2013, title={Long-Term Follow-up of Foamy Viral Vector-Mediated Gene Therapy for Canine Leukocyte Adhesion Deficiency}, volume={21}, ISSN={1525-0016}, url={http://dx.doi.org/10.1038/MT.2013.34}, DOI={10.1038/MT.2013.34}, abstractNote={The development of leukemia following gammaretroviral vector-mediated gene therapy for X-linked severe combined immunodeficiency disease and chronic granulomatous disease (CGD) has emphasized the need for long-term follow-up in animals treated with hematopoietic stem cell gene therapy. In this study, we report the long-term follow-up (4–7 years) of four dogs with canine leukocyte adhesion deficiency (CLAD) treated with foamy viral (FV) vector-mediated gene therapy. All four CLAD dogs previously received nonmyeloablative conditioning with 200 cGy total body irradiation followed by infusion of autologous, CD34+ hematopoietic stem cells transduced by a FV vector expressing canine CD18 from an internal Murine Stem Cell Virus (MSCV) promoter. CD18+ leukocyte levels were >2% following infusion of vector-transduced cells leading to ongoing reversal of the CLAD phenotype for >4 years. There was no clinical development of lymphoid or myeloid leukemia in any of the four dogs and integration site analysis did not reveal insertional oncogenesis. These results showing disease correction/amelioration of disease in CLAD without significant adverse events provide support for the use of a FV vector to treat children with leukocyte adhesion deficiency type 1 (LAD-1) in a human gene therapy clinical trial. The development of leukemia following gammaretroviral vector-mediated gene therapy for X-linked severe combined immunodeficiency disease and chronic granulomatous disease (CGD) has emphasized the need for long-term follow-up in animals treated with hematopoietic stem cell gene therapy. In this study, we report the long-term follow-up (4–7 years) of four dogs with canine leukocyte adhesion deficiency (CLAD) treated with foamy viral (FV) vector-mediated gene therapy. All four CLAD dogs previously received nonmyeloablative conditioning with 200 cGy total body irradiation followed by infusion of autologous, CD34+ hematopoietic stem cells transduced by a FV vector expressing canine CD18 from an internal Murine Stem Cell Virus (MSCV) promoter. CD18+ leukocyte levels were >2% following infusion of vector-transduced cells leading to ongoing reversal of the CLAD phenotype for >4 years. There was no clinical development of lymphoid or myeloid leukemia in any of the four dogs and integration site analysis did not reveal insertional oncogenesis. These results showing disease correction/amelioration of disease in CLAD without significant adverse events provide support for the use of a FV vector to treat children with leukocyte adhesion deficiency type 1 (LAD-1) in a human gene therapy clinical trial.}, number={5}, journal={Molecular Therapy}, publisher={Elsevier BV}, author={Bauer, Thomas R, Jr and Tuschong, Laura M and Calvo, Katherine R and Shive, Heather R and Burkholder, Tanya H and Karlsson, Eleanor K and West, Robert R and Russell, David W and Hickstein, Dennis D}, year={2013}, month={May}, pages={964–972} }
 @article{wei_simpson_johann_dwyer_prieto_kumar_ye_luke_shive_webster_et al._2012, title={Proteomic Profiling of H-Ras-G12V Induced Hypertrophic Cardiomyopathy in Transgenic Mice Using Comparative LC-MS Analysis of Thin Fresh-Frozen Tissue Sections}, volume={11}, ISSN={1535-3893 1535-3907}, url={http://dx.doi.org/10.1021/pr200612y}, DOI={10.1021/pr200612y}, abstractNote={Determination of disease-relevant proteomic profiles from limited tissue specimens, such as pathological biopsies and tissues from small model organisms, remains an analytical challenge and a much needed clinical goal. In this study, a transgenic mouse disease model of cardiac-specific H-Ras-G12V induced hypertrophic cardiomyopathy provided a system to explore the potential of using mass spectrometry (MS)-based proteomics to obtain a disease-relevant molecular profile from amount-limited specimens that are routinely used in pathological diagnosis. Our method employs a two-stage methanol-assisted solubilization to digest lysates prepared from 8-μm-thick fresh-frozen histological tissue sections of diseased/experimental and normal/control hearts. Coupling this approach with a nanoflow reversed-phase liquid chromatography (LC) and a hybrid linear ion trap/Fourier transform-ion cyclotron resonance MS resulted in the identification of 704 and 752 proteins in hypertrophic and wild-type (control) myocardium, respectively. The disease driving H-Ras protein along with vimentin were unambiguously identified by LC-MS in hypertrophic myocardium and cross-validated by immunohistochemistry and western blotting. The pathway analysis involving proteins identified by MS showed strong association of proteomic data with cardiovascular disease. More importantly, the MS identification and subsequent cross-validation of Wnt3a and β-catenin, in conjunction with IHC identification of phosphorylated GSK-3β and nuclear localization of β-catenin, provided evidence of Wnt/β-catenin canonical pathway activation secondary to Ras activation in the course of pathogenic myocardial hypertrophic transformation. Our method yields results indicating that the described proteomic approach permits molecular discovery and assessment of differentially expressed proteins regulating H-Ras induced hypertrophic cardiomyopathy. Selected proteins and pathways can be further investigated using immunohistochemical techniques applied to serial tissue sections of similar or different origin.}, number={3}, journal={Journal of Proteome Research}, publisher={American Chemical Society (ACS)}, author={Wei, Bih-Rong and Simpson, R. Mark and Johann, Donald J., Jr. and Dwyer, Jennifer E. and Prieto, DaRue A. and Kumar, Mia and Ye, Xiaoying and Luke, Brian and Shive, Heather R. and Webster, Joshua D. and et al.}, year={2012}, month={Feb}, pages={1561–1570} }
 @article{pepperberg_shive_2001, title={Simultaneous development of vocal and physical object combinations by a grey parrot (Psittacus erithacus): Bottle caps, lids, and labels}, volume={115}, DOI={10.1037//0735-7036.115.4.376-384}, abstractNote={On the basis of primarily behavioral data, researchers (e.g., P. M. Greenfield, 1991) have argued (a) that parallel development of communicative and physical object (manual) combinatorial abilities exists in young children; (b) that these abilities initially have a common neural substrate; (c) that a homologous substrate in great apes allows for similar, if limited, parallel development of these 2 abilities; and (d) that such abilities thus may indicate a shared evolutionary history for both communicative and physical behavior (J. Johnson-Pynn, D. M. Fragaszy, E. M. Hirsh, K. E. Brakke, & P. M. Greenfield, 1999). The authors of the present study found a comparable, if limited, parallel combinatorial development in a Grey parrot (Psittacus erithacus). Given the evolutionary distance between parrots and primates, the authors suggest that the search for and arguments concerning responsible substrates and common behavior should be approached with care and should not be restricted to the primate line.}, number={4}, journal={Journal of Comparative Pathology}, author={Pepperberg, I. M. and Shive, H. R.}, year={2001}, pages={376–384} }