van Winkoop A, Timmermans L P
Department of Experimental Animal Morphology and Cell Biology, Agricultural University, Wageningen, The Netherlands.
Histochemistry. 1992 Dec;98(5):289-98. doi: 10.1007/BF00270012.
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.
本文描述了一种从鲤鱼睾丸中分离大型早期精原细胞的方法，并通过在连续Percoll梯度上进行平衡密度离心，将其从最初4-5%的回收率纯化至60-70%。通过脾内途径用精原细胞免疫小鼠。选择并进一步分析了六种产生与生殖细胞选择性反应的单克隆抗体（MAb）的杂交瘤培养物。在增殖开始时，即在7周龄时，观察到其中五种MAb与发育中未分化性腺中的原始生殖细胞（PGC）发生反应。一种编码为WCG6的MAb似乎定义了PGC上一种新的表面标志物，该标志物在2至4周龄时在表面膜上逐渐表达，同时这些有丝分裂静止细胞的大小增加。在精子发生过程中，生殖细胞与其中五种MAb的反应完全消失（WCG 7、12、15、21）或几乎完全消失（WCG 6），这与先前针对鲤鱼精子产生的MAb所获得的模式形成了显著差异。在性腺发育过程中，用WCG-MAb未观察到雄性和雌性生殖细胞之间的差异，这表明在精原细胞和卵原细胞之前及包括它们在内的两性生殖细胞之间共享一组共同的表面抗原。
