title: Identification of novel genes expressed during spermatogenesis in stage-synchronized rat testes by differential display.
abstract: The molecular mechanism regulating spermatogenesis at different developmental stages remains largely unknown. In a vitamin A-deficiency (VAD) rat model, five distinct histologically defined, stage-synchronized testes: (i) resting spermatogonia and preleptotene spermatocytes at Day 0 of post-vitamin A treatment (PVA); (ii) early pachytene spermatocytes at Day 7 PVA; (iii) late pachytene at Day 15 PVA; (iv) round spermatids at Day 25 PVA; and (v) elongated spermatids at Day 35 PVA were used to study gene expression profiles by mRNA differential display. Twenty-four differentially expressed cDNA fragments were identified and cloned; oligonucleotide sequence analyses indicated that there are 12 novel gene sequences, half of which share no apparent match in current GenBank/EMBL databases. Other 12 VAD clones share sequence homology to membrane channel and transport, transcription and translation, cell cycle and morphogenesis, inducer and transducer, surface or secreted glycoproteins or enzymes, and other miscellaneous molecules. Semi-quantitative RT-PCR analyses against different stages of VAD testes demonstrated: (i) restricted expression of VAD1.2 and 1.3 (novel) on Day 25 PVA when round spermatids form; (ii) escalating pattern of VAD12 (Cx43) in Sertoli cells; and (iii) relative constant levels of VAD4 (A5D3), VAD26.1 (ribonuclease), and VAD27 (GRP8) in spermatogenesis.
authors.affiliation: Department of Surgery, University of Hong Kong Medical Center, Hong Kong, Special Administrative Region of China, PR China. jmluk@hkucc.hku.hk
authors.forename: John M
authors.initials: JM
authors.lastname: Luk
authors.forename: B W
authors.initials: BW
authors.lastname: Mok
authors.forename: C K
authors.initials: CK
authors.lastname: Shum
authors.forename: W S
authors.initials: WS
authors.lastname: Yeung
authors.forename: P C
authors.initials: PC
authors.lastname: Tam
authors.forename: J Y
authors.initials: JY
authors.lastname: Tse
authors.forename: J F
authors.initials: JF
authors.lastname: Chow
authors.forename: J
authors.initials: J
authors.lastname: Woo
authors.forename: K
authors.initials: K
authors.lastname: Kam
authors.forename: K F
authors.initials: KF
authors.lastname: Lee
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000302:isolation & purification; D020869:Gene Expression Profiling; D007700:Kinetics; D008297:Male; D009928:Organ Specificity; D051381:Rats; D017207:Rats, Sprague-Dawley; D013091:Spermatogenesis / Q000235:genetics*; D013737:Testis / Q000033:anatomy & histology / Q000166:cytology / Q000378:metabolism*; D014802:Vitamin A Deficiency / Q000235:genetics / Q000378:metabolism
chemical_list: D018076:DNA, Complementary