Den Boer P J, Grootegoed J A
Department of Biochemistry II, Erasmus University Rotterdam, The Netherlands.
J Reprod Fertil. 1988 Jul;83(2):701-9. doi: 10.1530/jrf.0.0830701.
Tubular fragments (spermatogenic epithelium) from immature hamsters were isolated and cultured with low doses of (+)- and (-)-gossypol enantiomers. The activity of lactate dehydrogenase isoenzyme LDH-C4 was estimated as a marker for spermatogenic cell development and alpha-ketoisovalerate was used as the substrate. In the absence of gossypol, the specific activity of LDH-C4 in the tubular fragments was increased 40% during incubation for 48 h. This developmental increase was suppressed by gossypol. The specific activity of LDH-C4 in the tubular fragments was lowered by gossypol, after 48 h of culture in the presence of low doses of racemic gossypol (1-4 microM) and 1% fetal calf serum. In this in-vitro system the (-)-enantiomer but not the (+)-enantiomer of gossypol affected the LDH-C4 activity. This is in agreement with other reports showing that only the (-)-enantiomer induces infertility. The observed action of gossypol on LDH-C4 activity in the tubular fragments may reflect gossypol-induced degeneration of spermatogenic cells. The present in-vitro system can be used to estimate the actions of gossypol derivatives, other investigational antifertility agents, and toxic agents on the spermatogenic epithelium.
从未成熟仓鼠中分离出管状片段(生精上皮),并用低剂量的(+)-和(-)-棉酚对映体进行培养。乳酸脱氢酶同工酶LDH-C4的活性被用作生精细胞发育的标志物进行评估,α-酮异戊酸被用作底物。在没有棉酚的情况下,管状片段中LDH-C4的比活性在孵育48小时期间增加了40%。这种发育性增加被棉酚抑制。在低剂量消旋棉酚(1-4 microM)和1%胎牛血清存在的情况下培养48小时后,棉酚降低了管状片段中LDH-C4的比活性。在这个体外系统中,棉酚的(-)-对映体而非(+)-对映体影响了LDH-C4的活性。这与其他表明只有(-)-对映体诱导不育的报道一致。观察到的棉酚对管状片段中LDH-C4活性的作用可能反映了棉酚诱导的生精细胞退化。目前的体外系统可用于评估棉酚衍生物、其他研究性抗生育药物和有毒物质对生精上皮的作用。