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The expression and localisation of mRNAs for 5 alpha reductase Type I (5 alpha R-I) and Type II (5 alpha R-II) isoenzymes in human benign prostatic hyperplasia (BPH) were investigated by RT-PCR and by in mini hybridisation (ISH) using digoxigenin labelled riboprobes. In addition, we also examined the isoenzymes mRNA expression in primary BPH cultures of separated stroma/fibroblast and epithelial cells to determine whether primary cultures are appropriate models in which to investigate 5 alpha R activity and regulation. The results demonstrated conclusively the presence of mRNA encoding both isoenzymes in all specimens so far examined. Additionally, the presence of a functional 5 alpha R-I and -II activity in BPH was confirmed by enzyme assays. ISH studies localised the mRNA expression to both the fibroblast/stromal component as well as the epithelial cells of the hyperplastic tissue. In the glandular regions the expression for both isoenzymes was particularly strong in the basal layers of the epithelium whereas mRNA expression in the secretory cells was less pronounced. Expression of 5 alpha R-I and -II mRNAs in fibroblast was on the other hand variable with high expression in some areas and little in others. These findings were supported by our primary culture experiments which demonstrated that both the fibroblast and epithelial cells maintain a capacity to express both isoenzymes in vitro. In the case of the fibroblast, the capacity to express the isoenzymes was maintained following the sequential passaging of the cells up to passage 6, after which the cells no longer expressed either isoenzyme.
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ABSTRACT
The effect of altering the number of pups per litter from ten to two or two to ten at day 14 of lactation in rats was investigated. Reducing litter size had no effect on the daily live weight gain whereas increasing the litter size resulted in an initial weight loss followed by an impaired weight gain. Maternal food consumption decreased to values appropriate for rats feeding two pups within 2 days after litter reduction, but required at least 5 days after the increase in litter size to increase to values appropriate for rats feeding large litters from parturition. Altering the litter size resulted in maternal serum insulin concentrations that were intermediate between those expected for rats feeding two or ten pups from parturition. Maternal serum prolactin concentrations increased after litter sizes were increased and decreased initially after litter reduction before being restored to normal levels. The activities of the lipogenic enzymes, fatty acid synthase, glucose-6-phosphate dehydrogenase and 'malic' enzyme in the mammary gland all decreased within 72 h of litter reduction to levels appropriate for rats feeding two pups. Although all three enzymes increased in activity after litter size increase, only fatty acid synthase had reached values appropriate for rats feeding ten pups by 72 h.
J. Endocr. (1984) 101, 69–73
