SPERMATID METABOLISM SYMBIOSIS WITH SERTOLI CELL

建议扩展对精子细胞新陈代谢的早期研究，以与 该细胞对Sertoli“哺乳”功能的作用 细胞。一种以胶原为基质的支持细胞培养体系的建立 为了允许支持细胞在体外呈现其真实的柱状形状，因此 两个隔室(基底室和侧腔室)可以发育，细胞可以 区别于塑料表面上看到的扁平形式。这个 细胞将被灌流到专门为此目的而设计的白榴石室中。这个 细胞的经典特性(形态、蛋白质合成--包括 雄激素结合蛋白、核酸合成、环磷酸腺苷和 乳酸盐，以及对激素的反应)将被检查以比较 在常规培养系统中培养支持细胞。我们会调查的 离子梯度特征产生的机制 生精小管的活体(高钾，低钠 隔室)，以及激素对梯度的影响。自然界和 分泌到侧脑室的蛋白质的数量将是 下定决心。最后，将尝试对Sertoli细胞进行共培养 将生殖细胞(精原细胞；精母细胞)引入基底部 车厢。光学显微镜和电子显微镜(包括延时显微镜 电影摄影)，将被用于检测生殖细胞的分化 (精母细胞-精子细胞)与残余体的发生 遵循共同文化。生殖细胞将用(~3H)胸腺嘧啶核苷标记 体内的通过遗传标记来区分细胞和供体大鼠加入 受体支持细胞来自贴壁的污染生殖细胞 受体大鼠。

It is proposed to extend earlier studies on spermatid metabolism to relate the functions of this cell to the so-called "nursing" function of Sertoli cells. A system for culturing Sertoli cells on collagen has been developed to permit the Sertoli cell to assume its true columnar shape in vitro so that two compartments (basal and adluminal) can develop and the cell can differentiate beyond the flattened form seen on plastic surfaces. The cells will be perfused in a leucite chamber designed for the purpose. The classical properties of the cell (morphology, protein synthesis - including androgen-binding protein, synthesis of nucleic acid, cyclic AMP and lactate, together with responses to hormones) will be examined to compare with Sertoli cells in the conventional culture system. We will investigate the mechanisms involved in production of the ionic gradients characteristic of the seminiferous tubule in vivo (high K+; low Na+ in adluminal compartment), and the effect of hormones on the gradient. The nature and number of proteins secreted into the adluminal compartment will be determined. Finally, attempts will be made to co-culture Sertoli cells with germ cells (spermatogonia; spermatocytes) introduced into the basal compartment. Light and electron microscopy (including time-lapse cinematography), will be used to detect differentiation of germ cells (spermatocytes - spermatids) and the occurrence of residual bodies following co-culture. Germ cells will be labeled with (3H) thymidine in vivo of by genetic markers to distinguish cells from donor rats added to recipient Sertoli cells from adherent contaminating germ cells from the recipient rat.