Feig L A, Bellvé A R, Erickson N H, Klagsbrun M
Proc Natl Acad Sci U S A. 1980 Aug;77(8):4774-8. doi: 10.1073/pnas.77.8.4774.
The seminiferous epithelium of the mouse testis contains a potent mitogen that induces DNA synthesis and cell division in cutures of confluent, quiescent BALB/c 3T3 cells. Homogenates of adult mouse seminiferous tubules, when added to 3T3 cells for 48 hr, produce a dose-dependent increase in DNA synthesis with half-maximal activity at 400 micrograms of protein per ml. Maximal stimulation causes a 70- to 100-fold increase in [methyl-3H]thymidine incorporation into DNA, a labeling of > 99% of 3T3 cell nuclei, and a doubling in cell number. The level of mitogenic activity in seminiferous tubules isolated from newborn mice is five times greater than that in tubules of adult mice. Furthermore, prepuberal Sertoli cells contain substantially greater levels of mitogenic activity than any other cell type purified from the seminiferous epithelium. A homogenate of prepuberal Sertoli cells induces half-maximal DNA synthesis at 20 micrograms of protein per ml. The mitogenic factor is sensitive to proteases and to heating at 100 degrees C for 2 min but not to dithiothreitol. Gel-filtration chromatography reveals that the factor is a polypeptide with a Mr of approximately 15,500. These observations provide evidence for the presence of a growth factor in the mammalian testis.
小鼠睾丸的生精上皮含有一种强效促有丝分裂原,可在汇合、静止的BALB/c 3T3细胞培养物中诱导DNA合成和细胞分裂。成年小鼠生精小管的匀浆加入到3T3细胞中48小时后,会引起DNA合成呈剂量依赖性增加,每毫升400微克蛋白质时达到半数最大活性。最大刺激可使[甲基-3H]胸腺嘧啶掺入DNA增加70至100倍,使>99%的3T3细胞核被标记,并使细胞数量增加一倍。从新生小鼠分离的生精小管中的促有丝分裂活性水平比成年小鼠小管中的高五倍。此外,青春期前的支持细胞所含的促有丝分裂活性水平比从生精上皮纯化的任何其他细胞类型都高得多。青春期前支持细胞的匀浆在每毫升20微克蛋白质时可诱导半数最大DNA合成。该促有丝分裂因子对蛋白酶和在100℃加热2分钟敏感,但对二硫苏糖醇不敏感。凝胶过滤色谱显示该因子是一种分子量约为15,500的多肽。这些观察结果为哺乳动物睾丸中存在生长因子提供了证据。