Griswold M D, Mably E R, Fritz I B
Mol Cell Endocrinol. 1976 Feb;4(3):139-49. doi: 10.1016/0303-7207(76)90033-2.
The incorporation of [2H]thymidine into nuclear DNA was investigated in cultured Sertoli cells prepared from testes of 20-day-old rats. Addition of follicle-stimulating hormone (FSH) or dibutyryl cyclic, 3',5'-adenosine monophosphate (DBCAMP) to the culture medium greatly increased incorporation, expressed either as total amounts of [3H]thymidine incorporated per mug DNA or as the percentage of Sertoli cells with labeled nuclear DNA. No stimulation was observed in cells cultured in the presence of testosterone, insulin or cyclic 3',5'-GMP (cGMP). Light and electron microscopic autoradiographic analysis was employed to establish the identity of Sertoli cells having labeled nuclear DNA. Contaminating spermatogonia, which also took up labeled [3H]thymidine, were excluded from cell counts. In addition, Sertoli cells prepared from testes of irradiated 20-day-old germinal cell depleted rats were also observed to incorporate more [3H]thymidine into nuclear DNA when cultured in a chemically defined medium in the presence of FSH. DNA synthesis was abolished by prior treatment of cells with cytosine arabinoside. In separate experiments, the incorporation of [3H]thymidine into DNA of peritubular myoid cells was shown to be independent of FSH or dbcAMP.
在从20日龄大鼠睾丸制备的培养支持细胞中,研究了[2H]胸苷掺入核DNA的情况。向培养基中添加促卵泡激素(FSH)或二丁酰环3',5'-单磷酸腺苷(DBCAMP)可显著增加掺入量,掺入量可表示为每微克DNA中掺入的[3H]胸苷总量,也可表示为具有标记核DNA的支持细胞的百分比。在睾酮、胰岛素或环3',5'-鸟苷单磷酸(cGMP)存在下培养的细胞中未观察到刺激作用。采用光镜和电镜放射自显影分析来确定具有标记核DNA的支持细胞的身份。细胞计数中排除了也摄取标记[3H]胸苷的污染精原细胞。此外,当在化学限定培养基中于FSH存在下培养时,观察到从受辐照的20日龄生精细胞耗竭大鼠的睾丸制备的支持细胞也将更多的[3H]胸苷掺入核DNA中。用阿糖胞苷预先处理细胞可消除DNA合成。在单独的实验中,显示[3H]胸苷掺入睾丸肌样细胞DNA与FSH或dbcAMP无关。