RhoA/Rho-kinase suppresses endothelial nitric oxide synthase in the penis

A mechanism for diabetes-associated erectile dysfunction

Trinity J. Bivalacqua, Hunter C. Champion, Mustafa F. Usta, Selim Cellek, Kanchan Chitaley, R Clinton Webb, Ronald W Lewis, Thomas M. Mills, Wayne J G Hellstrom, Philip J. Kadowitz

Research output: Contribution to journalArticle

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Abstract

Significant impairment in endothelial-derived nitric oxide is present in the diabetic corpus cavernosum. RhoA/Rho-kinase may suppress endothelial nitric oxide synthase (eNOS). Here, we tested the hypothesis that RhoA/Rho-kinase contributes to diabetes-related erectile dysfunction and down-regulation of eNOS in the streptozotocin (STZ)-diabetic rat penis. Colocalization of Rho-kinase and eNOS protein was present in the endothelium of the corpus cavernosum. RhoA/Rho-kinase protein abundance and MYPT-1 phosphorylation at Thr-696 were elevated in the STZ-diabetic rat penis. In addition, eNOS protein expression, cavernosal constitutive NOS activity, and cGMP levels were reduced in the STZ-diabetic penis. To assess the functional role of RhoA/Rho-kinase in the penis, we evaluated the effects of an adeno-associated virus encoding the dominant-negative RhoA mutant (AAVTCMV19NRhoA) on RhoA/Rho-kinase and eNOS and erectile function in vivo in the STZ-diabetic rat. STZ-diabetic rats transfected with AAVCMVT19NRhoA had a reduction in RhoA/Rho-kinase and MYPT-1 phosphorylation at a time when cavernosal eNOS protein, constitutive NOS activity, and cGMP levels were restored to levels found in the control rats. There was a significant decrease in erectile response to cavernosal nerve stimulation in the STZ-diabetic rat. AAVT19NRhoA gene transfer improved erectile responses in the STZ-diabetic rat to values similar to control. These data demonstrate a previously undescribed mechanism for the down-regulation of penile eNOS in diabetes mediated by activation of the RhoA/Rho-kinase pathway. Importantly, these data imply that inhibition of RhoA/Rho-kinase improves eNOS protein content and activity thus restoring erectile function in diabetes.

Original languageEnglish (US)
Pages (from-to)9121-9126
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume101
Issue number24
DOIs
StatePublished - Jun 15 2004

Fingerprint

rho-Associated Kinases
Nitric Oxide Synthase Type III
Penis
Erectile Dysfunction
Streptozocin
Proteins
Down-Regulation
Phosphorylation
Dependovirus
Endothelium
Nitric Oxide

Keywords

  • Endothelium
  • Gene therapy
  • Neuronal NO synthase

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

RhoA/Rho-kinase suppresses endothelial nitric oxide synthase in the penis : A mechanism for diabetes-associated erectile dysfunction. / Bivalacqua, Trinity J.; Champion, Hunter C.; Usta, Mustafa F.; Cellek, Selim; Chitaley, Kanchan; Webb, R Clinton; Lewis, Ronald W; Mills, Thomas M.; Hellstrom, Wayne J G; Kadowitz, Philip J.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 101, No. 24, 15.06.2004, p. 9121-9126.

Research output: Contribution to journalArticle

Bivalacqua, Trinity J. ; Champion, Hunter C. ; Usta, Mustafa F. ; Cellek, Selim ; Chitaley, Kanchan ; Webb, R Clinton ; Lewis, Ronald W ; Mills, Thomas M. ; Hellstrom, Wayne J G ; Kadowitz, Philip J. / RhoA/Rho-kinase suppresses endothelial nitric oxide synthase in the penis : A mechanism for diabetes-associated erectile dysfunction. In: Proceedings of the National Academy of Sciences of the United States of America. 2004 ; Vol. 101, No. 24. pp. 9121-9126.
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AU - Chitaley, Kanchan

AU - Webb, R Clinton

AU - Lewis, Ronald W

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