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

• Published in: Biology of reproduction 1996-10, Vol.55 (4), p.923-932
• Main Authors: WOLKOWICZ, M. J, COONROD, S. M, PRABHAKARA REDDI, P, MILLAN, J. L, HOFMANN, M.-C, HERR, J. C
• Format: Article
• Language: English
• Subjects: Acrosin - analysis; Acrosin - genetics; Acrosome; Animal cells; Animals; Antigens; Antigens, Differentiation - analysis; Antigens, Differentiation - genetics; Antigens, Differentiation - immunology; Antigens, Surface - analysis; Antigens, Surface - genetics; Biological and medical sciences; Biotechnology; Blotting, Northern; Cell Differentiation - genetics; Cell Differentiation - immunology; Cell Differentiation - physiology; Cell Line, Transformed; Electrophoresis, Polyacrylamide Gel; Eukaryotic cell cultures; Flow Cytometry; Fundamental and applied biological sciences. Psychology; Genetical aspects; Gonadal Steroid Hormones; Humans; Immunoblotting; Isoenzymes; L-Lactate Dehydrogenase - analysis; L-Lactate Dehydrogenase - genetics; Male; Membrane Proteins; Methods. Procedures. Technologies; Mice; Phenotype; Polymerase Chain Reaction; Protamines - analysis; Protamines - genetics; Proteins - analysis; Proteins - genetics; RNA - analysis; RNA - genetics; Spermatocytes - cytology; Spermatocytes - physiology; Spermatogenesis - physiology; Temperature; Time Factors
• ISSN: 0006-3363; 1529-7268
• 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.