Gene homologues were identified by carrying out BLAST searches of the chicken genome and EST databases. Following identification, an expression profile of each gene was obtained using RT-PCR, Northern and <i>in situ </i>hybridisation analyses carried out on embryos at stages of embryonic development which PGC specification; PGC migration to the gonads and PGC development in the gonads occurred. Analyses focused on the genes <i>bruno, germ cell-less, oct4, mago nashi, nanog, nanos, piwi, pumilio </i>and <i>staufen </i>and aimed to determine (1) whether gene expression was comparable to the expression of homologues in other species, (2) whether the gene was expressed in areas known to contain PGCs, (3) whether the gene was expressed specifically in PGCs and (4) whether the gene could have a potential role in PGC development. Of the genes chosen to be investigated, homologues of <i>bruno, germ cell-less, mago nashi, piwi, pumilio </i>and <i>staufen </i>were identified in the genome and in EST databases. No <i>nanos </i>homologues were identified by this method. Homologues of <i>oct4 </i>and <i>nanog </i>were identified by Dr B.Pain who provided sequence data for both genes. analyses showed that the expression of all the genes were comparable to expression patterns in other species. <i>Nanog </i>and <i>oct4 </i>were expressed in a pattern that indicated germ cell specific expression. This led to additional <i>in situ</i> hybridisation, PGC transfer and qPCR analyses at later stages of development. These data showed how expression of <i>oct4 </i>and <i>nanog </i>changed in relation to (1) the loss of PGC motility in males and females and (2) the onset of meiosis in females. Of the remaining genes, <i>germ cell-less </i>and <i>piwi </i>were detected specifically in adult testes, whereas the rest were detected in areas of the embryo known to contain PGCs. However, the gene expression was not PGC specific.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:654399 |
| Date | January 2007 |
| Creators | MacLeod, David J. |
| Publisher | University of Edinburgh |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://hdl.handle.net/1842/17025 |
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