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Title: Characterisation of the receptor tyrosine kinase, Sek-1, during vertebrate embryogenesis
Author: Irving, Carol
ISNI:       0000 0001 3587 0901
Awarding Body: University of London
Current Institution: University College London (University of London)
Date of Award: 1997
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During segmentation of the vertebrate hindbrain expression of a number of genes is precisely restricted within the rhombomeres. One such gene is Sek-1, a member of the Eph subfamily of receptor tyrosine kinases, originally identified through its striking segmental expression pattern in both the rhombomeres and the presomitic mesoderm in the mouse. I raised antibodies against mouse Sek-1 protein that are specific for a protein of 110 KDa, and cross-react with a protein of identical size in chick, zebrafish and Xenopus. I used the anti-Sek antibodies to carry out a spatio-temporal analysis of the protein in comparison to the gene transcript. Cross-reactivity of the antibodies allowed a comparison of Sek-1 protein expression between species, revealing a high level of conservation of expression, but with some differences. Sek-1 RNA is detected in the presomitic mesoderm in two stripes immediately caudal to the latest somite formed, corresponding to rostral somitomeres. I found that Sek-1 protein is also expressed here, but expression persists in the newly formed somites, becoming apically localised in the epithelium. In the rhombomeres, Sek-1 protein and RNA expression domains arise in pre- r3 and pre-r5 in narrow stripes that broaden. Expression initially occurs in fuzzy domains that sharpen as the rhombomere boundaries form. I propose that the establishment and maintainance of the rhombomere segments may involve both a dynamic regulation of r3/r5 identity and the restriction of cell movement across rhombomere boundaries. I also carried out experiments to attempt to interfere with the function of Sek-1 in the hindbrain using transgenic approaches. I designed constructs for the ectopic expression of Sek-1 cDNA specifically in r4, and for the expression of a truncated form of Sek-1 in r3/5 in a dominant negative strategy to specifically inactivate Sek-1 in the hindbrain.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available
Keywords: Biochemistry