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Sara Lynn N Farwell, Kimberly G Reylander, M Kathryn Iovine, Linda J Lowe-Krentz
Transmembrane protein 184A (TMEM184A) was recently identified as the heparin receptor in vascular cells. Heparin binds specifically to TMEM184A and induces anti-proliferative signaling in vitro. Though it is highly conserved, the physiological function of TMEM184A remains unknown. The objective of this study was to investigate the expression and effects on vascular regeneration of TMEM184A using the adult zebrafish regenerating caudal fin as an in vivo model. Here, we show that Tmem184a is expressed in vascular endothelial cells (ECs) of mature and regenerating zebrafish fins...
2017: Frontiers in Physiology
Sara Lynn N Farwell, Joshua B Slee, Yaqiu Li, Linda J Lowe-Krentz
When novel proteins are identified through affinity-based isolation and bioinformatics analysis, they are often largely uncharacterized. Antibodies against specific peptides within the predicted sequence allow some localization experiments. However, other possible interactions with the antibodies often cannot be excluded. This situation provided an opportunity to develop a set of assays dependent on the protein sequence. Specifically, a construct containing the gene sequence coupled to the GFP coding sequence at the C-terminal end of the protein was obtained and employed for these purposes...
February 17, 2017: Journal of Visualized Experiments: JoVE
Soichiro Ogaki, Seiko Harada, Nobuaki Shiraki, Kazuhiko Kume, Shoen Kume
BACKGROUND: We developed an efficient in vitro method to differentiate mouse ES cells into the definitive endoderm (DE) and then Pdx1-expressing pancreatic lineages using mesodermal-derived supporting cells, M15. Using this method, resulting ES cell-derived DE and Pdx1-expressing cells were isolated by cell sorting, and their gene expression profiles were investigated with DNA microarray. Genes that were specifically expressed in DE and/or in Pdx1-expressing cells were extracted and their expression patterns in normal embryonic development were studied...
2011: BMC Developmental Biology
Terje Svingen, Dagmar Wilhelm, Alexander N Combes, Brett Hosking, Vincent R Harley, Andrew H Sinclair, Peter Koopman
Gene function during mouse development is often studied through the production and analysis of transgenic and knockout models. However, these techniques are time- and resource-consuming, and require specialized equipment and expertise. We have established a new protocol for functional studies that combines organ culture of explanted fetal tissues with microinjection and magnetically induced transfection ("magnetofection") of gene expression constructs. As proof-of-principle, we magnetofected cDNA constructs into genital ridge tissue as a means of gain-of-function analysis, and shRNA constructs for loss-of-function analysis...
April 2009: Developmental Dynamics: An Official Publication of the American Association of Anatomists
Terje Svingen, Annemiek Beverdam, Pascal Bernard, Peter McClive, Vincent R Harley, Andrew H Sinclair, Peter Koopman
During mouse embryogenesis, the fate of the bipotential gonads is sealed around 10.5 days post coitum (dpc) when the Y-linked gene Sry specifies the differentiation of testes in males, whereas in females, absence of Sry results in ovary formation. Apart from the pivotal action of Sry, many other genes are known to be involved in sex determination and subsequent differentiation. Much is still unknown regarding the regulatory hierarchy governing these events and many more sex differentiation genes are yet to be discovered...
May 2007: Reproduction: the Official Journal of the Society for the Study of Fertility
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