Davis J C, Himmelstein A, Bordy M, Desjardins C
Endocrinology. 1979 Dec;105(6):1419-25. doi: 10.1210/endo-105-6-1419.
Sertoli cells dissociated from 10-day-old rat testes form colonies in primary culture in response to FSH. FSH and dibutyryl cAMP stimulated the attachment of Sertoli cells to an equal extent; however, FSH-treated cultures contained a small number of large colonies while dibutyryl cAMP-treated cultures contained a large number of small colonies. This relationship was not altered by the addition of a number of other peptide or steroid hormones. Extracellular cAMP levels and colony density were negatively correlated. Colony size at 24 h of culture was diminished in FSH-treated cultures by the addition of a cAMP antibody at 6 or 12 h of incubation. The addition of cAMP at 24 h to FSH-treated cultures caused a dose-dependent stimulation of colony size but not colony density at 48 h of culture. A point source of cAMP (4 x 10(-3) M in agar) inhibited migration of cells toward the agar spot. An agar spot on the dish substratum containing Sepharose-bound FSH exhibited a halo of cells next to the spot, with a zone lacking cells distal to it. Radioautographs of [125I]iodo-FSH-treated cultures exhibited a nonhomogeneous distribution of silver grains; colony size increased faster than the number of labeled cells. Taken together, the results suggest that 1) FSH is stimulating the attachment of Sertoli cells through an increase in intracellular cAMP, 2) FSH is promoting active aggregation of Sertoli cells in culture through a modulation of extracellular cAMP, and 3) cells with a large amount of bound FSH are acting as centers for aggregation.
从10日龄大鼠睾丸中分离出的支持细胞在原代培养中对促卵泡激素(FSH)产生反应而形成集落。FSH和二丁酰环磷腺苷(dibutyryl cAMP)同等程度地刺激支持细胞的附着;然而,经FSH处理的培养物中含有少量大集落,而经二丁酰环磷腺苷处理的培养物中含有大量小集落。添加多种其他肽类或甾体激素不会改变这种关系。细胞外环磷腺苷水平与集落密度呈负相关。在培养6小时或12小时时添加环磷腺苷抗体,可使FSH处理的培养物在培养24小时时的集落大小减小。在培养24小时时向FSH处理的培养物中添加环磷腺苷,在培养48小时时可引起集落大小的剂量依赖性刺激,但不影响集落密度。环磷腺苷的点源(琼脂中4×10⁻³ M)抑制细胞向琼脂斑点的迁移。培养皿基质上含有与琼脂糖结合的FSH的琼脂斑点旁边有一圈细胞,其远端有一个无细胞区域。[¹²⁵I]碘-FSH处理的培养物的放射自显影片显示银粒分布不均匀;集落大小的增加比标记细胞的数量快。综合来看,结果表明:1)FSH通过增加细胞内环磷腺苷来刺激支持细胞的附着;2)FSH通过调节细胞外环磷腺苷促进培养中支持细胞的主动聚集;3)结合大量FSH的细胞作为聚集中心。