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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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