Refinement of the differentiated phenotype of the spermatogenic cell line GC-2spd(ts)

Abstract

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.