Phenotypic changes in germ cells during gonadal development of the common carp (Cyprinus carpio). An immunohistochemical study with anti-carp spermatogonia monoclonal antibodies. |
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Authors: | A van Winkoop L P Timmermans |
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Institution: | Department of Experimental Animal Morphology and Cell Biology, Agricultural University, Wageningen, The Netherlands. |
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Abstract: | A procedure is described in which large early spermatogonia were isolated from carp testes and purified from an initial 4-5% recovery up to 60-70% using equilibrium density centrifugation on a continuous Percoll gradient. Mice were immunized with the spermatogonia via the intrasplenic route. Six hybridoma cultures, producing monoclonal antibodies (MAbs) reacting selectively with germ cells, were selected and further analysed. Reactivity with five of these MAbs was observed on primordial germ cells (PGCs) in the developing indifferent gonads at the onset of proliferation, i.e. the age of 7 weeks. One MAb, encoded WCG6, appeared to define a new surface marker on PGCs being gradually expressed on the surface membrane between the age of 2 and 4 weeks, concomitantly with an increase in size of these mitotically silent cells. The reactivity of germ cells with five of the MAbs disappeared completely (WCG 7, 12, 15, 21) or nearly completely (WCG 6) during spermatogenesis, providing a striking difference from patterns obtained with MAbs raised previously against carp spermatozoa. Differences between male and female germ cells were not observed with the WCG-MAbs during gonad development, indicating that a common set of surface antigens is shared between germ cells of both sexes up to and including spermatogonia and oogonia. |
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