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Stage-Specific Regulation of Stem Cell Factor Gene Expression in the Rat Seminiferous Epithelium¹

Departments of Physiology and Pediatrics, University of Turku, Kiinamyllynkatu 10, Turku 20520, Finland

Address all correspondence and requests for reprints to: Dr. Jorma Toppari, Department of Physiology, University of Turku, Kiinamyllynkatu 10, Turku 20520, Finland. E-mail: jorma.toppari{at}utu.fi

To assess the regulation of stem factor factor (SCF) gene expression during spermatogenesis, we tested the effects of hormones (FSH, testosterone, and 17ß-estradiol) and some growth factors [transforming growth factor-ß (TGFß), TGF, tumor necrosis factor-, and activin] on SCF gene expression by using a transillumination-assisted microdisection technique, a seminiferous tubule culture system, and Northern hybridization. Our results showed that FSH (10 ng/ml) increased steady state levels of SCF messenger RNA (mRNA) in a stage-specific and time-dependent manner. 8-Bromo-cAMP could increase the SCF mRNA level in a similar way as FSH, whereas phorbol 12-myristate 13-acetate had no effect. Actinomycin D could abolish the stimulatory effect of FSH, whereas cyclohexamide could not. The half-life of SCF mRNA was apparently prolonged after FSH stimulation (FSH-treated tubules, 15.6 ± 1.2 h; controls, 8.6 ± 2.7 h). Nuclear run-on assay revealed 5- and 10-fold increases in the transcription rate after FSH stimulation for 8 and 30 h, respectively. Neither testosterone nor estradiol had significant effects on SCF gene expression in our tissue culture system. Activin, TGFß, TGF, and tumor necrosis factor- had no effect on SCF gene expression in vitro. In conclusion, SCF gene expression in the rat seminiferous tubule is regulated by FSH through the cAMP/protein kinase A pathway. FSH regulates SCF gene expression at both transcriptional and posttranscriptional levels involving the increase in transcription rate and prolongation of half-life of SCF mRNA, but is independent of de novo protein synthesis.

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