Biology of Reproduction, Vol 55, 923-932, Copyright © 1996 by Society for the Study of Reproduction

ARTICLES

Refinement of the differentiated phenotype of the spermatogenic cell line GC-2spd(ts) [published erratum appears in Biol Reprod 1996 Dec;55(6):1458]

MJ Wolkowicz, SA Coonrod, PP Reddi, JL Millan, MC Hofmann, JC Herr and SA] Coonrod SM$[corrected to Coonrod
Department of Cell Biology, University of Virginia, Charlottesville 22908, USA.

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.

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