Journal
BIOLOGY OF REPRODUCTION
Volume 63, Issue 3, Pages 704-714Publisher
OXFORD UNIV PRESS INC
DOI: 10.1095/biolreprod63.3.704
Keywords
gene regulation; glutamate; male sexual function; nitric oxide; penis
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Funding
- NIDDK NIH HHS [R01DK02568, R01DK53069] Funding Source: Medline
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Penile erection is mediated by nitric oxide (NO) synthesized by the neuronal nitric oxide synthase (nNOS). In the rat penis, the main nNOS mRNA variant, PnNOS, differs from cerebellar nNOS (CnNOS) by a 102 base pair insert encoding a 34-amino acid sequence, In the mouse, two nNOS mRNAs have been identified: nNOS alpha, encoding a 155-kDa protein, and an exon 2-deletion variant, nNOS beta, encoding a 135-kDa protein that lacks a domain where a protein inhibitor of nNOS (PIN) binds. We wished to determine whether PnNOS alpha and beta are expressed in the rat penis and are located in the nerves and whether the beta form persists in the potent nNOS knock-out mouse (nNOS(Delta Delta)). A PnNOS antibody against the insert common to both PnNOS alpha and beta detected the expected 155-kDa protein in PnNOS alpha-transfected cells. This antibody, and the one common to PnNOS/CnNOS, showed (on Western blots) the 155- and 135-kDa nNOS variants in rat penile tissue during development and aging. PnNOS alpha mRNA and its subvariants were found as the main nNOS in the penile corpora, the cavernosal nerve, and the pelvic ganglia, with lower levels of PnNOS beta mRNA. In tissue sections, PnNOS protein was immunodetected in the penile nerve endings in the rat and in the nNOS wild-type and nNOS(Delta Delta) mice. An antibody against the sequence encoded by exon 2 did not react (on Western blots) with the 135-kDa band, which confirms that this protein is the beta form. In conclusion, both PnNOS alpha and beta are expressed in the rat penis at all ages and are located in the nerves, The beta form may allow nitric oxide synthesis during erection to be partially insensitive to PIN. The residual expression of PnNOS, and possibly CnNOS, in the penis of the nNOS(Delta Delta) mouse occurs through transcription of the beta mRNA, and this may explain the retention of erectile function when the expression of nNOS alpha is disrupted.
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