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Browsing by Subject "microsatellite"

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  • Alitalo, Annamari (Helsingin yliopistoUniversity of HelsinkiHelsingfors universitet, 2003)
    Primordial germ cells are the embryonic precursor cells of sperm and ova. They are concluded to differentiate outside the gonadal area probably in the extraembryonic mesoderm. Their invasive migration through the stromal tissues of the dorsal mesentery adherent to the extracellular matrix is a well accepted fact in mice. The aim of this study was to set up a technique based on the use of individual-specific markers to test the hypothesis on the migration of primordial germ cells in a large animal model. The hypothesis, supported by feeble proof until now, states that there would be substantial chimerism in the progeny of primordial germ cells in bovine twins originating from transmission of the cells through placental vascular anastomoses during fetal life. Our aim was pursued by microsatellite analysis of the highly polymorphic bovine MHC locus DRB3. Polymerase chain reaction and capillary electrophoresis were optimized using 21 non-chimeric animals, ten cows and eleven bulls. The DRB3 microsatellite of one bull was cloned to isolate single alleles. Whole blood-derived DNA from 11 chimeric animals, the leukocyte chimerims of which had earlier been quantified by in situ hybridisation, and 47 semen-derived DNA samples from twin borne AI bulls were analysed in the last phase of this study. Primordial germ cell chimerism did not seem to occur in a substantial quantity in twin borne bovine bulls. Extension of the analysis described here to other polymorphic repeat loci should clarify the cases were traces of possible chimerism was found by analysing the above locus.