Stage-specific gene expression during rat spermatogenesis: application of the mRNA differential display method.
Spermatogenesis is a complex differentiation process which requires the coordinate synthesis of diverse stage-specific proteins. In attempting a large-scale identification and characterization of those proteins, we have made use of the recently described mRNA differential display method (Liang and Pardee, Science 257: 967-971, 1992). This method is based on the reverse transcription of mRNAs obtained from two different cell populations (pachytene spermatocytes and spermatids in the present study) followed by a PCR reaction and comparison of the individual cDNA populations in a polyacrylamide gel system. Up to the present we have been able to identify 268 cDNA bands. Most of them (77%) are common to both cell stages. From the differentially expressed bands (23%) an ample majority was spermatid-specific (74%). According to our present results we conclude that the mRNA differential display is a promising approach for investigations on stage-specific gene expression during a differentiation process like spermatogenesis.
Geisinger
A
A
Department of Cell and Developmental Biology, Theodor-Boveri-Institute (Biocenter), University of Würzburg, Germany.
Wettstein
R
R
Benavente
R
R
D000818:Animals; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D018507:Gene Expression Regulation, Developmental; D008297:Male; D008722:Methods; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics*; D051381:Rats; D017208:Rats, Wistar; D013087:Spermatids / Q000378:metabolism; D013090:Spermatocytes / Q000378:metabolism; D013091:Spermatogenesis / Q000235:genetics*; D014158:Transcription, Genetic
D018076:DNA, Complementary; D012333:RNA, Messenger