Wolkowicz M J, Coonrod S A, Reddi P P, Millan J L, Hofmann M C, Herr J C
Department of Cell Biology, University of Virginia, Charlottesville 22908, USA.
Biol Reprod. 1996 Oct;55(4):923-32. doi: 10.1095/biolreprod55.4.923.
A transformed spermatogenic cell line GC-2spd(ts), recently reported to express a protein marker of spermiogenesis, was tested for the presence of several mRNAs encoded by genes transcribed specifically in the testis and at precise stages of spermatogenesis. Northern blotting and reverse-transcriptase polymerase chain reaction techniques showed that mRNAs for the stage-specific marker proteins LDH-C4 (preleptotene), acrosin (premeiotic), protamine-2 (postmeiotic), and SP-10 (postmeiotic round spermatid stage) were not detected in GC-2spd(ts) cells. Flow cytometric analysis of GC-2spd(ts) failed to detect a peak indicative of the presence of haploid chromosomes. Furthermore, the HS-63 monoclonal antibody, employed in an earlier report to demonstrate putative proacrosomal granules, failed to recognize the SP-10 protein in extracts of human or mouse sperm or in GC-2spd(ts) cells and instead recognized proteins of different masses. In view of interest in this line as a model for analyzing molecular events of spermatogenesis, this refinement of the GC-2spd(ts) phenotype may aid others considering these cells for studies of terminal stages of sperm differentiation.
最近报道的一种转化的生精细胞系GC-2spd(ts)，据报道表达精子发生的一种蛋白质标志物，对其进行检测，以确定几种由在睾丸中特异性转录以及在精子发生的精确阶段转录的基因所编码的mRNA的存在情况。Northern印迹法和逆转录酶聚合酶链反应技术表明，在GC-2spd(ts)细胞中未检测到阶段特异性标志物蛋白乳酸脱氢酶C4（细线前期）、顶体蛋白酶（减数分裂前）、鱼精蛋白-2（减数分裂后）和SP-10（减数分裂后圆形精子细胞阶段）的mRNA。对GC-2spd(ts)的流式细胞术分析未能检测到表明存在单倍体染色体的峰值。此外，在早期报告中用于证明假定的前顶体颗粒的HS-63单克隆抗体，未能识别来自人或小鼠精子提取物或GC-2spd(ts)细胞中的SP-10蛋白，而是识别了不同质量的蛋白质。鉴于该细胞系作为分析精子发生分子事件模型的研究兴趣，GC-2spd(ts)表型的这种细化可能有助于其他考虑将这些细胞用于精子分化终末阶段研究的人员。
