Parker B Antin

PMID: 8647921;Abstract:

Endoderm within the heart forming regions of vertebrate embryos has pronounced effects on myocardial cell development. Previous studies have suggested that these effects are mediated by soluble growth factors, in particular fibroblast growth factor 2 (FGF-2) and activin-A. Since both insulin and insulin-like growth factors (IGFs) are present in developing avian embryos at the time of heart formation, we have investigated the potential role of these molecules in promoting development of premyocardial cells in quail. Culture of precardiac mesoderm explants from stage 5 quail embryos in medium containing insulin, IGF-I, or IGF-II increased proliferation of premyocardial cells, with maximal stimulation observed at approximately 25 nM for each ligand. A direct comparison of the proliferative response of precardiac mesoderm to endoderm, fetal calf serum, insulin, IGF-I, IGF-II, activin-A, and FGF-2 showed that FGF-2 and activin-A increased proliferation of premyocardial cells approximately 2-fold, while insulin, IGF-I, and IGF-II stimulated proliferation approximately 3-fold. Insulin and IGF-I enhanced the rate of myocyte differentiation, similar to previously reported effects of endoderm. In contrast, exposure of precardiac mesoderm explants to transforming growth factor beta (TGFβ) reduced proliferation of premyocardial cells and moderated the proliferative effects of IGF-I. TGFβ did not block the differentiation of stage 5 premyocardial cells. Reverse transcription-polymerase chain reaction (RT-PCR) analyses showed that mRNAs encoding insulin, IGF-II, insulin receptor, and IGF-I receptor were present in both precardiac mesoderm and endoderm, as well as in the forming heart at stage 8. Since premyocardial cells can survive and differentiate in a defined medium lacking these factors precardiac mesoderm may produce IGF-II and insulin at levels that are sufficient to stimulate myocyte development. Taken together, these results suggest that insulin and/or IGF-II may promote cardiac development in vivo by both autocrine and paracrine mechanisms. Cardiogenesis may therefore be promoted by the combined action of several classes of growth factors.

PMID: 19607656;PMCID: PMC2966335;Abstract:

Comparative genomics is an essential component of the post-genomic era. The chicken genome is the first avian genome to be sequenced and it will serve as a model for other avian species. Moreover, due to its unique evolutionary niche, the chicken genome can be used to understand evolution of functional elements and gene regulation in mammalian species. However comparative biology both within avian species and within amniotes is hampered due to the difficulty of recognising functional orthologs. This problem is compounded as different databases and sequence repositories proliferate and the names they assign to functional elements proliferate along with them. Currently, genes can be published under more than one name and one name sometimes refers to unrelated genes. Standardized gene nomenclature is necessary to facilitate communication between scientists and genomic resources. Moreover, it is important that this nomenclature be based on existing nomenclature efforts where possible to truly facilitate studies between different species. We report here the formation of the Chicken Gene Nomenclature Committee (CGNC), an international and centralized effort to provide standardized nomenclature for chicken genes. The CGNC works in conjunction with public resources such as NCBI and Ensembl and in consultation with existing nomenclature committees for human and mouse. The CGNC will develop standardized nomenclature in consultation with the research community and relies on the support of the research community to ensure that the nomenclature facilitates comparative and genomic studies. © 2009 Burt et al; licensee BioMed Central Ltd.

PMID: 10096065;Abstract:

In a screen for novel sequences expressed during embryonic heart development we have isolated a gene which encodes a putative RNA-binding protein. This protein is a member of one of the largest families of RNA- binding proteins, the RRM (RNA Recognition Motif) family. The gene has been named hermes (for HEart, RRM Expressed Sequence). The hermes protein is 197- amino acids long and contains a single RRM domain. In situ hybridization analysis indicates that hermes is expressed at highest levels in the myocardium of the heart and to a lesser extent in the ganglion layer of the retina, the pronephros and the epiphysis. Expression of hermes in each of these tissues begins at approximately the time of differentiation and is maintained throughout development analysis of the RNA expression of the hermes orthologues from chicken and mouse reveals that, like Xenopus, the most prominent tissue of expression is the developing heart. The sequence and expression pattern of hermes suggests a role in post-transcriptional regulation of heart development.

PMID: 19608613;PMCID: PMC3071757;Abstract:

Islet replacement is a promising therapy for treating diabetes mellitus, but the supply of donor tissue for transplantation is limited. To overcome this limitation, endocrine tissue can be expanded, but this requires an understanding of normal developmental processes that regulate islet formation. In this study, we compare pancreas development in sheep and human, and provide evidence that an epithelial-mesenchymal transition (EMT) is involved in β-cell differentiation and islet formation. Transcription factors know to regulate pancreas formation, pancreatic duodenal homeobox factor 1, neurogenin 3, NKX2-2, and NKX6-1, which were expressed in the appropriate spatial and temporal pattern to coordinate pancreatic bud outgrowth and direct endocrine cell specifi-cation in sheep. Immunofluorescence staining of the developing pancreas was used to co-localize insulin and epithelial proteins (cytokeratin, E-cadherin, and β-catenin) or insulin and a mesenchymal protein (vimentin). In sheep, individual β-cells become insulin-positive in the progenitor epithelium, then lose epithelial characteristics, and migrate out of the epithelial layer to form islets. As β-cells exit the epithelial progenitor cell layer, they acquire mesenchymal characteristics, shown by their acquisition of vimentin. In situ hybridization expression analysis of the SNAIL family members of transcriptional repressors (SNAIL1, -2, and -3; listed as SNAI1, -2, -3 in the HUGO Database) showed that each of the SNAIL genes was expressed in the ductal epithelium during development, and SNAIL-1 and -2 were co-expressed with insulin. Our findings provide strong evidence that the movement of β-cells from the pancreatic ductal epithelium involves an EMT. © 2009 Society for Endocrinology.

PMID: 20086052;PMCID: PMC2822927;Abstract:

In situ hybridization is widely used to visualize transcribed sequences in embryos, tissues, and cells. For whole mount detection of mRNAs in embryos, hybridization with an antisense RNA probe is followed by visual or fluorescence detection of target mRNAs. A limitation of this approach is that a cDNA template of the target RNA must be obtained in order to generate the antisense RNA probe. Here we investigate the use of short (12-24 nucleotides) locked nucleic acid (LNA) containing DNA probes for whole mount in situ hybridization detection of mRNAs. Following extensive protocol optimization, we show that LNA probes can be used to localize several mRNAs of varying abundances in chicken embryos. LNA probes also detected alternatively spliced exons that are processed in a tissue specific manner. The use of LNA probes for whole mount in situ detection of mRNAs will enable in silico design and chemical synthesis and will expand the general use of in situ hybridization for studies of transcriptional regulation and alternative splicing. Published by Cold Spring Harbor Laboratory Press. Copyright © 2010 RNA Society.