Functional Reconstitution of Vascular Smooth Muscle Cells With cGMP-Dependent Protein Kinase I Isoforms

Robert Feil; Nicolai Gappa; Mark Rutz; Jens Schlossmann; Christine R. Rose; Arthur Konnerth; Sabine Brummer; Susanne Kühbandner; Franz Hofmann

doi:10.1161/01.RES.0000019586.95768.40

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Functional Reconstitution of Vascular Smooth Muscle Cells With cGMP-Dependent Protein Kinase I Isoforms

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Functional Reconstitution of Vascular Smooth Muscle Cells With cGMP-Dependent Protein Kinase I Isoforms

Robert Feil, Nicolai Gappa, Mark Rutz, Jens Schlossmann, Christine R. Rose, Arthur Konnerth, Sabine Brummer, Susanne Kühbandner et Franz Hofmann

Circulation research, Vol.90(10), pp.1080-1086

31/05/2002

DOI: https://doi.org/10.1161/01.RES.0000019586.95768.40

PMID: 12039797

Résumé

The cGMP-dependent protein kinase type I (cGKI) is a major mediator of NO/cGMP-induced vasorelaxation. Smooth muscle expresses two isoforms of cGKI, cGKIα and cGKIβ, but the specific role of each isoform in vascular smooth muscle cells (VSMCs) is poorly understood. We have used a genetic deletion/rescue strategy to analyze the functional significance of cGKI isoforms in the regulation of the cytosolic Ca 2+ concentration by NO/cGMP in VSMCs. Cultured mouse aortic VSMCs endogenously expressed both cGKIα and cGKIβ. The NO donor diethylamine NONOate (DEA-NO) and the membrane-permeable cGMP analogue 8-bromo-cGMP inhibited noradrenaline-induced Ca 2+ transients in wild-type VSMCs but not in VSMCs genetically deficient for both cGKIα and cGKIβ. The defective Ca 2+ regulation in cGKI-knockout cells could be rescued by transfection of a fusion construct consisting of cGKIα and enhanced green fluorescent protein (EGFP) but not by a cGKIβ-EGFP construct. Fluorescence imaging indicated that the cGKIα-EGFP fusion protein was concentrated in the perinuclear/endoplasmic reticulum region of live VSMCs, whereas the cGKIβ-EGFP protein was more homogeneously distributed in the cytoplasm. These results suggest that one component of NO/cGMP-induced smooth muscle relaxation is the activation of the cGKIα isoform, which decreases the noradrenaline-stimulated cytosolic Ca 2+ level.

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Titre: Functional Reconstitution of Vascular Smooth Muscle Cells With cGMP-Dependent Protein Kinase I Isoforms
Créateurs - sans rôle: Robert Feil - From the Institut für Pharmakologie und Toxikologie der Technischen Universität München (R.F., N.G., M.R., J.S., S.B., S.K., F.H.) and Physiologisches Institut (C.R.R., A.K.), Universität München, München, Germany
Nicolai Gappa - From the Institut für Pharmakologie und Toxikologie der Technischen Universität München (R.F., N.G., M.R., J.S., S.B., S.K., F.H.) and Physiologisches Institut (C.R.R., A.K.), Universität München, München, Germany
Mark Rutz - From the Institut für Pharmakologie und Toxikologie der Technischen Universität München (R.F., N.G., M.R., J.S., S.B., S.K., F.H.) and Physiologisches Institut (C.R.R., A.K.), Universität München, München, Germany
Jens Schlossmann - From the Institut für Pharmakologie und Toxikologie der Technischen Universität München (R.F., N.G., M.R., J.S., S.B., S.K., F.H.) and Physiologisches Institut (C.R.R., A.K.), Universität München, München, Germany
Christine R. Rose - From the Institut für Pharmakologie und Toxikologie der Technischen Universität München (R.F., N.G., M.R., J.S., S.B., S.K., F.H.) and Physiologisches Institut (C.R.R., A.K.), Universität München, München, Germany
Arthur Konnerth - From the Institut für Pharmakologie und Toxikologie der Technischen Universität München (R.F., N.G., M.R., J.S., S.B., S.K., F.H.) and Physiologisches Institut (C.R.R., A.K.), Universität München, München, Germany
Sabine Brummer - From the Institut für Pharmakologie und Toxikologie der Technischen Universität München (R.F., N.G., M.R., J.S., S.B., S.K., F.H.) and Physiologisches Institut (C.R.R., A.K.), Universität München, München, Germany
Susanne Kühbandner - From the Institut für Pharmakologie und Toxikologie der Technischen Universität München (R.F., N.G., M.R., J.S., S.B., S.K., F.H.) and Physiologisches Institut (C.R.R., A.K.), Universität München, München, Germany
Franz Hofmann - Physiologisches Institut
Détails de publication: Circulation research, Vol.90(10), pp.1080-1086
Nombre de pages: 7
Identifiants: 99153943309311
Unité académique: Institut de Génétique Moléculaire de Montpellier - IGMM
Langue: English
Type de ressource: Journal article