Expression of prohibitin in rat seminiferous epithelium.
Subtractive hybridization was used to isolate cDNAs highly expressed in stages IX-XI of the cycle of the seminiferous epithelium in the rat. One of the cloned cDNAs was sequenced and shown to be homologous to a previously described cDNA encoding rat prohibitin. Northern blot analyses showed that 1.9- and 1.2-kb transcripts were present in Sertoli cells whereas 1.5-, 1.2-, and 0.7-kb transcripts were expressed in germ cells. Western blot analyses with anti-peptide antibody to prohibitin revealed only a single 30-kDa protein in testis. Immunocytochemistry demonstrated that prohibitin protein was expressed constitutively in adult Leydig cells and Sertoli cells at all stages. Immunoreactivity of prohibition was very low in preleptotene spermatocytes, very high in leptotene spermatocytes, and very low in zygotene spermatocytes. In pachytene spermatocytes, immunoreactivity was very high in stages VII-XI and was minimal during stages XII and XIV. No protein was detected in spermatogonia and spermatocytes undergoing mitotic and meiotic divisions, respectively. These studies show that the prohibitin gene is expressed differentially in testis. The expression pattern of the prohibitin gene in rat testis appears to correlate with a proposed antiproliferative role of prohibitin.
Department of Biochemistry and Biophysics, Washington State University, Pullman 99164-4660.
N M
NM
Choongkittaworn
K H
KH
Kim
D B
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Danner
M D
MD
Griswold
D000375:Aging; D000818:Animals; D015152:Blotting, Northern; D002478:Cells, Cultured; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification; D015870:Gene Expression; D007150:Immunohistochemistry; D008297:Male; D009693:Nucleic Acid Hybridization; D000091402:Prohibitins; D011506:Proteins / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012097:Repressor Proteins; D012670:Seminiferous Epithelium / Q000378:metabolism*; D012708:Sertoli Cells / Q000737:chemistry / Q000378:metabolism; D013087:Spermatids / Q000737:chemistry; D013090:Spermatocytes / Q000737:chemistry; D013737:Testis / Q000737:chemistry / Q000378:metabolism
D000091402:Prohibitins; D011506:Proteins; D012333:RNA, Messenger; D012097:Repressor Proteins; D004247:DNA