The phosphorylation state of eNOS modulates vascular reactivity and outcome of cerebral ischemia in vivo
Dmitriy N. Atochin, … , William C. Sessa, Paul L. Huang
Dmitriy N. Atochin, … , William C. Sessa, Paul L. Huang
Published July 2, 2007
Citation Information: J Clin Invest. 2007;117(7):1961-1967. https://doi.org/10.1172/JCI29877.
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The phosphorylation state of eNOS modulates vascular reactivity and outcome of cerebral ischemia in vivo

Abstract

NO plays critical roles in vascular function. We show that modulation of the eNOS serine 1179 (S1179) phosphorylation site affects vascular reactivity and determines stroke size in vivo. Transgenic mice expressing only a phosphomimetic (S1179D) form of eNOS show greater vascular reactivity, develop less severe strokes, and have improved cerebral blood flow in a middle cerebral artery occlusion model than mice expressing an unphosphorylatable (S1179A) form. These results provide a molecular mechanism by which multiple diverse cardiovascular risks, such as diabetes and obesity, may be centrally integrated by eNOS phosphorylation in vivo to influence blood flow and cardiovascular disease. They also demonstrate the in vivo relevance of posttranslational modification of eNOS in vascular function.

Authors

Dmitriy N. Atochin, Annie Wang, Victor W.T. Liu, Jeffrey D. Critchlow, Ana Paula V. Dantas, Robin Looft-Wilson, Takahisa Murata, Salvatore Salomone, Hwa Kyoung Shin, Cenk Ayata, Michael A. Moskowitz, Thomas Michel, William C. Sessa, Paul L. Huang

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Figure 1

Generation of S1179A and S1179D eNOS transgenic mice.

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Generation of S1179A and S1179D eNOS transgenic mice. (A) Transgenic con...
(A) Transgenic construct. A 1.6-kb fragment of the human eNOS promoter drives the bovine eNOS cDNA carrying the S1179D or S1179A mutation, an HA tag, and the SV40 polyadenylation site. (B) Genotyping of transgenic and native eNOS genes by PCR. Primers E1 and E2 (see Methods) span an intron, so they amplify an 800-bp product from the endogenous WT eNOS gene but a 280-bp product from the S1179A and S1179D transgenes. Mice bred onto the homozygous eNOS KO background do not show the 800-bp product. (C) Western blot analysis of heart protein (60 μg) of WT, S1179D transgenic/eNOS KO (S1179D/eNOS KO), and S1179A transgenic/eNOS KO (S1179A/eNOS KO) mice, using antibody directed against total eNOS.