Gerton G L, Millette C F
Biol Reprod. 1986 Nov;35(4):1025-35. doi: 10.1095/biolreprod35.4.1025.
Little is known about the ability of mammalian spermatogenic cells to synthesize plasma membrane components in the presence or absence of Sertoli cells. In this study, purified populations (greater than 90%) of pachytene spermatocytes or round spermatids were isolated by unit gravity sedimentation and cultured for 20-24 h in the presence of [35S]methionine or [3H]fucose. Cell viabilities remained over 90% during the course of these experiments. Plasma membranes were purified from these cells and analyzed by two-dimensional gel electrophoresis. Qualitatively, the same plasma membrane proteins were synthesized by both cell types with the exception of the major Concanavalin A-binding glycoprotein, p151; the synthesis of p151 is greatly diminished or inhibited after meiosis. [3H]Fucose was incorporated into at least 6 common glycoproteins of both cells. Eight components fucosylated with molecular weights from 35,000 to 120,000 were specific to pachytene spermatocyte membranes. One fast-migrating fucosylated component may represent an uncharacterized lipid whose synthesis is terminated after meiosis. Round spermatids specifically fucosylated two components with molecular weights of 45,000 and 80,000. These results demonstrate the viability of germ cells of the male mouse in short-term culture and show that they are capable of synthesizing and fucosylating plasma membrane components in the absence of Sertoli cells.
关于哺乳动物生精细胞在有或没有支持细胞的情况下合成质膜成分的能力，人们了解甚少。在本研究中，通过单位重力沉降法分离出纯化群体（大于90%）的粗线期精母细胞或圆形精子细胞，并在[35S]甲硫氨酸或[3H]岩藻糖存在的情况下培养20 - 24小时。在这些实验过程中，细胞活力保持在90%以上。从这些细胞中纯化质膜，并通过二维凝胶电泳进行分析。定性地说，除了主要的伴刀豆球蛋白A结合糖蛋白p151外，两种细胞类型合成的质膜蛋白相同；减数分裂后，p151的合成大大减少或受到抑制。[3H]岩藻糖被掺入到两种细胞的至少6种常见糖蛋白中。八种分子量从35,000到120,000的岩藻糖基化成分是粗线期精母细胞质膜特有的。一种快速迁移的岩藻糖基化成分可能代表一种未鉴定的脂质，其合成在减数分裂后终止。圆形精子细胞特异地使两种分子量分别为45,000和80,000的成分岩藻糖基化。这些结果证明了雄性小鼠生殖细胞在短期培养中的活力，并表明它们在没有支持细胞的情况下能够合成和岩藻糖基化质膜成分。
