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SPERMATID METABOLISM SYMBIOSIS WITH SERTOLI CELL

精子细胞代谢与支持细胞的共生

基本信息

批准号：
3313728
负责人：
PETER F HALL
金额：
$ 15.18万
依托单位：
WORCESTER FOUNDATION FOR BIOMEDICAL RES
依托单位国家：
美国
项目类别：
财政年份：
1983
资助国家：
美国
起止时间：
1983-02-01 至 1986-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3313728
关键词：
Sertoli cells androgens autoradiography cell differentiation cell growth regulation cell population study cyclic AMP electrofocusing electron microscopy follicle stimulating hormone gel electrophoresis genetic markers hormone regulation /control mechanism intercellular connection radiotracer sperm spermatogenesis tissue /cell culture

项目摘要

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.

建议扩展早期对精子细胞代谢的研究，这种细胞的功能与塞尔托利的所谓“护理”功能细胞建立了一种在胶原上培养睾丸支持细胞的系统使支持细胞在体外呈现真正的柱状，两个隔室（基底和近腔）可以发展，细胞可以区别于塑料表面上的扁平形式。的将细胞灌注在为此目的设计的白榴石室中。的细胞的经典特性（形态、蛋白质合成-包括雄激素结合蛋白，核酸合成，环腺苷酸，乳酸盐，以及对激素的反应）进行检查，在常规培养系统中培养支持细胞。我们将调查离子梯度产生的机制生精小管近腔高K+、低Na+ 隔室），以及激素对梯度的影响。的性质和分泌到近腔室中的蛋白质的数量将是测定最后，将尝试共培养支持细胞有生殖细胞（精原细胞;精母细胞）被引入基底层车厢光学和电子显微镜（包括延时电影摄影），将用于检测生殖细胞的分化（精母细胞-精子细胞）和残留体的发生共培养后。生殖细胞将用（3 H）胸苷标记，通过遗传标记来区分来自供体大鼠的细胞，受体支持细胞来自粘附的污染生殖细胞，受体大鼠