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The role of receptor tyrosine kinases in the control of early mammalian development
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Dissertation
The role of receptor tyrosine kinases in the control of early mammalian development
1995
Overview
A major aim of the vertebrate embryologist is to understand the mechanisms by which a group of cells controls the developmental fate of neighbouring cells. In this thesis, I have undertaken a study of the roles that growth factor signalling pathways play in the regulation of murine gastrulation and the formation and patterning of the embryonic mesoderm. A polymerase chain reaction screen was used to amplify tyrosine kinase mRNA expressed in in vitro-differentiated embryonic stem (ES) cells to initially define the growth factor receptor tyrosine kinases (RTKs) expressed during mammalian mesoderm formation. Fourteen different cytoplasmic or receptor tyrosine kinases were identified. Novel RTKs belonging to the Insulin, Ret, Kit/Pdgf, and FGF receptor families were isolated. Characterization centred around members of the FGFR and Kit/Pdgfr families of RTKs due to their demonstrated roles in a number of developmental systems. Both whole-mount and standard in situ hybridization analysis revealed that members of the Fgfr family were expressed early in development and suggested that they may play roles in controlling the development and segmentation of specific germ layers. The expression patterns of a Kit/Pdgfr-related RTK, now known as Flk1, and a novel RTK, designated Tek, correlate with the determination and differentiation, respectively, of the endothelial cell lineage. This work has identified the earliest known markers of endothelial cell precursors, blood vessels and the endocardium and has paved the way for a genetic and biochemical analysis of vasculogenesis. Functional characterization of Fgfr1 was carried out by gene targeting methodology. In the absence of Fgfr1 signalling, embryos displayed early growth defects, however they remained capable of gastrulating and generating mesoderm. The nascent mesoderm of Fgfr1 homozygous mutant embryos differentiated into diverse mesodermal subtypes, but mesodermal patterning was aberrant. Somites were never generated and axial mesoderm was greatly expanded at the expense of paraxial mesoderm. Embryos did not survive past embryonic day 9.5. These results suggest that FGFR1 transduces signals that specify mesodermal cell fates and regional patterning of the mesoderm during gastrulation and highlights the essential roles that growth signalling pathways play during embryogenesis.
Publisher
ProQuest Dissertations & Theses
Subject
ISBN
0612074099, 9780612074095
