Ueda H, Ziomek C, Hall P F
Worcester Foundation for Experimental Biology, Shrewsbury, Massachusetts 01545.
Endocrinology. 1988 Aug;123(2):1014-22. doi: 10.1210/endo-123-2-1014.
Sertoli cells were prepared from fetal rats, aged 15, 18, and 21 days. Cultures of these cells can be prepared at a purity of 85% by a method that is widely used to prepare the same cells from postnatal rats. The fetal cells are identical in appearance to postnatal Sertoli cells. Fetal Sertoli cells round up in response to (Bu)2cAMP, but not in response to FSH. Protein synthesis in the cells is not stimulated by (Bu)2cAMP or FSH. However, incorporation of [35S]methionine into secreted proteins appearing in the incubation medium is stimulated by the cyclic nucleotide, but not by FSH. Reproducible patterns for the incorporation of [35S]methionine into cellular and secreted proteins are presented as autoradiograms of one- and two-dimensional electrophoretograms. These autoradiograms show that the response of secreted proteins to (Bu)2cAMP is a general one; most or all proteins participate in the response. No clear differences were observed in the nature of the proteins synthesized when cells from fetal rats of 15, 18, and 21 days of age were compared. Fetal Sertoli cells produced approximately 2 nmol lactate/10(6) cells.h, which is approximately one fifth of the amount produced by postnatal cells. Lactate production was increased by the addition of FSH or (Bu)2cAMP to the medium. Fetal Sertoli cells also synthesize androgen-binding protein, and this activity is not increased by either FSH or (Bu)2cAMP.
支持细胞取自15、18和21日龄的胎鼠。这些细胞的培养物可以通过一种广泛用于从出生后大鼠制备相同细胞的方法,以85%的纯度制备。胎细胞在外观上与出生后支持细胞相同。胎支持细胞对(Bu)2cAMP有反应而变圆,但对促卵泡激素(FSH)无反应。细胞中的蛋白质合成不受(Bu)2cAMP或FSH的刺激。然而,环核苷酸可刺激[35S]甲硫氨酸掺入孵育培养基中出现的分泌蛋白,但FSH则不能。[35S]甲硫氨酸掺入细胞和分泌蛋白的可重复模式以一维和二维电泳图的放射自显影片呈现。这些放射自显影片表明,分泌蛋白对(Bu)2cAMP的反应是普遍的;大多数或所有蛋白质都参与了该反应。比较15、18和21日龄胎鼠的细胞时,在合成蛋白质的性质上未观察到明显差异。胎支持细胞每10(6)个细胞·小时产生约2 nmol乳酸,约为出生后细胞产生量的五分之一。向培养基中添加FSH或(Bu)2cAMP可增加乳酸的产生。胎支持细胞还合成雄激素结合蛋白,且该活性不受FSH或(Bu)2cAMP的增加。