Increased PDZ-RhoGEF/RhoA/Rho kinase signaling in small mesenteric arteries of angiotensin II-induced hypertensive rats

Rob H P Hilgers, Joseph Todd, R Clinton Webb

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

BACKGROUND: The phosphorylation of myosin light chain (MLC) maintains the contracted state of vascular smooth muscle. Dephosphorylation results in relaxation and is determined by the activity of myosin light chain phosphatase (MLCP), which is negatively regulated by Rho kinase. METHODS: We tested whether an increased Rho kinase activity, and hence a decreased contribution of MLCP, results in an increased contractility of small fourth-order mesenteric arteries (MA) during the early onset of angiotensin II (Ang II)-induced hypertension (Ang II-14d). RESULTS: Calcium sensitivity was similar, but contractile tension in response to [Ca]ex (5 mmol/l) in endothelium-denuded and depolarized MA was greater, in Ang II-14d rats compared to sham-operated normotensive (SHAM) and Ang II-1d. The Rho kinase inhibitor Y-27632 caused a significantly greater inhibition of the contractile response to various agents (phenylephrine, norepinephrine, U46619 and K) in MA of Ang II-14d compared to SHAM. Protein expression levels of the GDP/GTP exchange factor PDZ-RhoGEF, which co-immunoprecipitated with RhoA, were increased in MA of Ang II-14d compared to SHAM. RhoA translocation was greater in U46619 (1 μmol/l)-stimulated MA of Ang II-14d compared to SHAM. Expression levels of Rho kinase β were higher in MA of Ang II-14d. The MLCP inhibitor calyculin A (100 nmol/l) caused a greater contraction in MA of SHAM compared to Ang II-14d. Phosphorylation of the target subunit of MLCP (MYPT1) was enhanced in U46619-stimulated MA of Ang II-14d compared to SHAM. CONCLUSION: This is the first study demonstrating enhanced PDZ-RhoGEF/RhoA/Rho kinase signaling during hypertension at the level of resistance-sized arteries. This enhanced signaling leads to increased MLCP phosphorylation, resulting in vascular hyper-reactivity.

Original languageEnglish (US)
Pages (from-to)1687-1697
Number of pages11
JournalJournal of Hypertension
Volume25
Issue number8
DOIs
StatePublished - Aug 1 2007

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Rho Guanine Nucleotide Exchange Factors
rho-Associated Kinases
Mesenteric Arteries
Angiotensin II
Myosin-Light-Chain Phosphatase
15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid
Phosphorylation
Hypertension
Guanine Nucleotide Exchange Factors
Myosin Light Chains
Phenylephrine
Guanosine Triphosphate
Vascular Smooth Muscle
Endothelium
Blood Vessels
salicylhydroxamic acid
Norepinephrine
Arteries

Keywords

  • Angiotensin II
  • Calcium-sensitization
  • Hypertension
  • Mesenteric artery
  • Rho kinase

ASJC Scopus subject areas

  • Internal Medicine
  • Endocrinology

Cite this

Increased PDZ-RhoGEF/RhoA/Rho kinase signaling in small mesenteric arteries of angiotensin II-induced hypertensive rats. / Hilgers, Rob H P; Todd, Joseph; Webb, R Clinton.

In: Journal of Hypertension, Vol. 25, No. 8, 01.08.2007, p. 1687-1697.

Research output: Contribution to journalArticle

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T1 - Increased PDZ-RhoGEF/RhoA/Rho kinase signaling in small mesenteric arteries of angiotensin II-induced hypertensive rats

AU - Hilgers, Rob H P

AU - Todd, Joseph

AU - Webb, R Clinton

PY - 2007/8/1

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N2 - BACKGROUND: The phosphorylation of myosin light chain (MLC) maintains the contracted state of vascular smooth muscle. Dephosphorylation results in relaxation and is determined by the activity of myosin light chain phosphatase (MLCP), which is negatively regulated by Rho kinase. METHODS: We tested whether an increased Rho kinase activity, and hence a decreased contribution of MLCP, results in an increased contractility of small fourth-order mesenteric arteries (MA) during the early onset of angiotensin II (Ang II)-induced hypertension (Ang II-14d). RESULTS: Calcium sensitivity was similar, but contractile tension in response to [Ca]ex (5 mmol/l) in endothelium-denuded and depolarized MA was greater, in Ang II-14d rats compared to sham-operated normotensive (SHAM) and Ang II-1d. The Rho kinase inhibitor Y-27632 caused a significantly greater inhibition of the contractile response to various agents (phenylephrine, norepinephrine, U46619 and K) in MA of Ang II-14d compared to SHAM. Protein expression levels of the GDP/GTP exchange factor PDZ-RhoGEF, which co-immunoprecipitated with RhoA, were increased in MA of Ang II-14d compared to SHAM. RhoA translocation was greater in U46619 (1 μmol/l)-stimulated MA of Ang II-14d compared to SHAM. Expression levels of Rho kinase β were higher in MA of Ang II-14d. The MLCP inhibitor calyculin A (100 nmol/l) caused a greater contraction in MA of SHAM compared to Ang II-14d. Phosphorylation of the target subunit of MLCP (MYPT1) was enhanced in U46619-stimulated MA of Ang II-14d compared to SHAM. CONCLUSION: This is the first study demonstrating enhanced PDZ-RhoGEF/RhoA/Rho kinase signaling during hypertension at the level of resistance-sized arteries. This enhanced signaling leads to increased MLCP phosphorylation, resulting in vascular hyper-reactivity.

AB - BACKGROUND: The phosphorylation of myosin light chain (MLC) maintains the contracted state of vascular smooth muscle. Dephosphorylation results in relaxation and is determined by the activity of myosin light chain phosphatase (MLCP), which is negatively regulated by Rho kinase. METHODS: We tested whether an increased Rho kinase activity, and hence a decreased contribution of MLCP, results in an increased contractility of small fourth-order mesenteric arteries (MA) during the early onset of angiotensin II (Ang II)-induced hypertension (Ang II-14d). RESULTS: Calcium sensitivity was similar, but contractile tension in response to [Ca]ex (5 mmol/l) in endothelium-denuded and depolarized MA was greater, in Ang II-14d rats compared to sham-operated normotensive (SHAM) and Ang II-1d. The Rho kinase inhibitor Y-27632 caused a significantly greater inhibition of the contractile response to various agents (phenylephrine, norepinephrine, U46619 and K) in MA of Ang II-14d compared to SHAM. Protein expression levels of the GDP/GTP exchange factor PDZ-RhoGEF, which co-immunoprecipitated with RhoA, were increased in MA of Ang II-14d compared to SHAM. RhoA translocation was greater in U46619 (1 μmol/l)-stimulated MA of Ang II-14d compared to SHAM. Expression levels of Rho kinase β were higher in MA of Ang II-14d. The MLCP inhibitor calyculin A (100 nmol/l) caused a greater contraction in MA of SHAM compared to Ang II-14d. Phosphorylation of the target subunit of MLCP (MYPT1) was enhanced in U46619-stimulated MA of Ang II-14d compared to SHAM. CONCLUSION: This is the first study demonstrating enhanced PDZ-RhoGEF/RhoA/Rho kinase signaling during hypertension at the level of resistance-sized arteries. This enhanced signaling leads to increased MLCP phosphorylation, resulting in vascular hyper-reactivity.

KW - Angiotensin II

KW - Calcium-sensitization

KW - Hypertension

KW - Mesenteric artery

KW - Rho kinase

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