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Androgens Modulate Nitric Oxide Synthase Messenger Ribonucleic Acid Expression in Neurons of the Major Pelvic Ganglion in the Rat

Laboratoire de Neurobiologie des Fonctions Végétatives (A.S., C.M.), Laboratoire de Physiologie Sensorielle (C.B.), Laboratoire de Biologie Cellulaire et Moléculaire (E.D.), Institut National de la Recherche Agronomique, F-78352 Jouy-en-Josas Cedex, France

Address all correspondence and requests for reprints to: Dr. Alain Schirar, Laboratoire de Neurobiologie des Fonctions Végétatives, INRA, F-78352 Jouy-en-Josas Cedex, France. E-mail: schirar{at}jouy.inra.fr

Expression and androgen regulation of the gene for neuronal nitric oxide synthase (NOS I) were examined in neurons of the major pelvic ganglia in male rats. Some of these postganglionic neurons innervate the penis and produce nitric oxide, which is believed to play a major role in penile erection. Rats were either castrated or sham operated and implanted with SILASTIC brand capsules filled with powdered testosterone (T) or 5-dihydrotestosterone (5DHT) or left empty. After 4 days, the number of neurons intensely stained for NADPH-diaphorase as well as those giving a NOS I signal in in situ hybridization experiments increased in castrated rats treated with testosterone by 31% and 42%, respectively, relative to those in untreated castrated rats. This suggests that the increase in NADPH-diaphorase activity resulted from enzyme synthesis and was due to a modification of NOS I messenger RNA (mRNA) accumulation. After 7 days, Northern blot analysis showed that castration produced a decrease in the amount of NOS I mRNA relative to that of ribosomal RNA. This decrease was almost prevented by T treatment. No significant differences were observed by reverse transcriptase-PCR between 7-day and 28-day treatments. However, in 7-day castrated rats treated with 5DHT, NOS I signals relative to those of hypoxanthine phosphoribosyltransferase, taken as reference, were significantly higher than those in castrated rats and resembled those in sham-castrated rats, suggesting that 5DHT was probably more potent than testosterone in preventing the decrease in NOS I mRNA levels elicited by castration. These results show that NOS I can be positively regulated by androgens and are consistent with the suggestion that these steroids play a role in the physiological processes of penile erection.

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