Pem homeobox gene promoter sequences that direct transcription in a Sertoli cell-specific, stage-specific, and androgen-dependent manner in the testis in vivo.
Although many genes are expressed selectively in Sertoli cells, regulatory sequences sufficient to drive Sertoli cell-specific expression in the postnatal and adult testis in vivo have not been identified. In the present study, we identified promoter sequences from the Pem homeobox gene that direct Sertoli cell-specific expression in an androgen-dependent and stage-specific manner. Immunohistochemical and RNA analysis demonstrated that 0.6-kb 5'-flanking sequence directed transgene expression specifically in the testis and the epididymis but not in any other tissues tested. In the adult testis, this promoter fragment targeted the transgene expression specifically to Sertoli cells during stages IV-VIII of the seminiferous epithelial cycle, thereby mimicking the expression pattern of the endogenous Pem gene. This promoter fragment also recapitulated Pem's normal postnatal expression pattern, as it directed transcript induction between d 6 and d 9 post partum. Deletion of 0.3 kb from the 5'-end of the transgene had no effect on androgen-dependent Sertoli-specific expression but altered stage-specific expression in adult testes and caused premature postnatal expression. Our results suggest that there are at least two regulatory regions in the Pem proximal promoter: one that directs androgen receptor-dependent expression specifically in Sertoli cells within the testis and another that confers stage-specific expression in neonates and adults by acting as a negative regulator. To our knowledge, this is the first identification of regulatory regions that direct faithful developmentally regulated gene expression in postnatal and adult Sertoli cells in vivo.
Rao
Manjeet K
MK
Department of Immunology, The University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030, USA.
Wayne
Chad M
CM
Meistrich
Marvin L
ML
Wilkinson
Miles F
MF
D024506:5' Flanking Region; D000726:Androgen Antagonists / Q000494:pharmacology; D000728:Androgens / Q000378:metabolism*; D000818:Animals; D002478:Cells, Cultured; D005485:Flutamide / Q000494:pharmacology; D018507:Gene Expression Regulation, Developmental; D005801:Genes, Homeobox; D049452:Green Fluorescent Proteins; D018398:Homeodomain Proteins / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009928:Organ Specificity; D011401:Promoter Regions, Genetic; D012708:Sertoli Cells / Q000166:cytology / Q000502:physiology*; D013737:Testis / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D013739:Testosterone / Q000378:metabolism; D014157:Transcription Factors / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
D000726:Androgen Antagonists; D000728:Androgens; D005485:Flutamide; D049452:Green Fluorescent Proteins; D018398:Homeodomain Proteins; D008164:Luminescent Proteins; D013739:Testosterone; D014157:Transcription Factors; C498048:Rhox5 protein, mouse