A 3-kilobase region derived from the rat cathepsin L gene directs in vivo expression of a reporter gene in sertoli cells in a manner comparable to that of the endogenous gene.
During mammalian spermatogenesis, the transcription of several genes in Sertoli cells is turned on and off as the adjacent male germ cells progress through the stages of the cycle of the seminiferous epithelium. A requirement for defining how germ cells regulate this process is the identification of a promoter that confers, in vivo, accurate, stage-specific gene expression in Sertoli cells. To date, such a promoter has not been identified. Using transgenic mice, we show that the 3-kilobase genomic fragment immediately upstream of the rat cathepsin L translation start site directs expression of the reporter gene, beta-galactosidase, only in Sertoli cells. The expression pattern of the reporter gene recapitulated that of the endogenous gene in Sertoli cells as 75% of the seminiferous tubules that contained X-gal positive Sertoli cells were at stages VI-VIII and beta-galactosidase enzymatic activity was 4-fold higher in mature testes compared with immature testes. This is, to our knowledge, the first identification of a promoter region that contains all of the regulatory elements required for accurate, stage-specific gene expression in Sertoli cells.
Charron
Martin
M
Division of Reproductive Biology, Department of Biochemistry and Molecular Biology, Bloomberg School of Public Health, The Johns Hopkins University, Baltimore, Maryland 21205, USA.
Folmer
Janet S
JS
Wright
William W
WW
D000818:Animals; D056668:Cathepsin L; D002403:Cathepsins / Q000235:genetics*; D003546:Cysteine Endopeptidases; D004247:DNA / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics*; D007763:Lac Operon / Q000235:genetics; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D014176:Protein Biosynthesis / Q000235:genetics; D012670:Seminiferous Epithelium / Q000378:metabolism; D012671:Seminiferous Tubules / Q000201:enzymology; D012708:Sertoli Cells / Q000378:metabolism*; D019076:Transgenes / Q000235:genetics; D001616:beta-Galactosidase / Q000378:metabolism
D004247:DNA; D001616:beta-Galactosidase; D002403:Cathepsins; D003546:Cysteine Endopeptidases; D056668:Cathepsin L; C534278:Ctsl protein, mouse; C534280:Ctsl protein, rat