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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 Gene¹

Division of Reproductive Biology,³ Department of Biochemistry and Molecular Biology, Bloomberg School of Public Health, The Johns Hopkins University, Baltimore, Maryland 21205

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, ß-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 ß-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.

¹ Supported in part by the NICHHD, NIH (through Cooperative Agreement U54-HD-36209), as part of the Specialized Cooperative Centers Program in Reproduction Research. Additional support was provided by the NIH (RO1-HD-17989) and the Hopkins Population Center (P30-HD-06268).

² Correspondence: William W. Wright, Johns Hopkins University Bloomberg School of Public Health, Department of Biochemistry and Molecular Biology, Room 3508, 615 North Wolfe St., Baltimore, MD 21205. FAX: 410 614 2356; wwright1{at}jhem.jhmi.edu

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