Refinement of the differentiated phenotype of the spermatogenic cell line GC-2spd(ts)
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.
Wolkowicz
M J
MJ
Department of Cell Biology, University of Virginia, Charlottesville 22908, USA.
Coonrod
S A
SA
Reddi
P P
PP
Millan
J L
JL
Hofmann
M C
MC
Herr
J C
JC
Coonrod
S M
SM
D000176:Acrosin / Q000032:analysis / Q000235:genetics; D000177:Acrosome; D000818:Animals; D000941:Antigens; D000943:Antigens, Differentiation / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D000954:Antigens, Surface / Q000032:analysis / Q000235:genetics; D015152:Blotting, Northern; D002454:Cell Differentiation / Q000235:genetics / Q000276:immunology / Q000502:physiology; D002461:Cell Line, Transformed; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D012739:Gonadal Steroid Hormones; D006801:Humans; D015151:Immunoblotting; D007527:Isoenzymes; D007770:L-Lactate Dehydrogenase / Q000032:analysis / Q000235:genetics; D008297:Male; D008565:Membrane Proteins; D051379:Mice; D010641:Phenotype; D016133:Polymerase Chain Reaction; D011479:Protamines / Q000032:analysis / Q000235:genetics; D011506:Proteins / Q000032:analysis / Q000235:genetics; D012313:RNA / Q000032:analysis / Q000235:genetics; D013090:Spermatocytes / Q000166:cytology* / Q000502:physiology; D013091:Spermatogenesis / Q000502:physiology*; D013696:Temperature; D013997:Time Factors
C512729:ACRV1 protein, human; C080034:Acrv1 protein, mouse; D000941:Antigens; D000943:Antigens, Differentiation; D000954:Antigens, Surface; D012739:Gonadal Steroid Hormones; D007527:Isoenzymes; D008565:Membrane Proteins; D011479:Protamines; D011506:Proteins; C068621:protamine 2; D012313:RNA; D007770:L-Lactate Dehydrogenase; D000176:Acrosin