Internalization and dephosphorylation of connexin43 in hypertrophied right ventricles of rats with pulmonary hypertension.
BACKGROUND
Altered expression and distribution of gap junctions might provide substrates for abnormal conduction and arrhythmogenesis in the heart, but little is known about the regulation of gap junctions under pathological conditions. The organization and phosphorylation state of connexin43 (Cx43) in ventricular hypertrophy will be investigated.


RESULTS
Right ventricular (RV) hypertrophy was induced in rats by treatment with monocrotaline. Subcellular Cx43 distribution was assessed by immunoconfocal and electron microscopy. Immunolabeling of Cx43 was confined to the intercalated disks in the normal ventricular myocytes of control rats, but hypertrophied RV cells from monocrotaline-treated rats showed dispersion of Cx43 immunolabeling over the cell surface and in the cytoplasm; cytoplasmic Cx43 was increased by approximately 7-fold (n=15). The Cx43 internalization was confirmed by the double staining of monocrotaline-treated RV tissues for Cx43/wheat germ agglutinin (WGA) and Cx43/zonula occludens protein-1 (ZO-1). Electron microscopy of hypertrophied RVs showed an increase in annular gap junctions immunolabeled with Cx43. Immunoblotting revealed a significant increase in non-phosphorylated Cx43 in hypertrophied RVs (by approximately 5-fold, n=8) without changes in the total amount of Cx43. The accumulation of non-phosphorylated Cx43 in hypertrophied RVs was also recognized by immunoconfocal-microscopy with an isoform-specific antibody.


CONCLUSIONS
Ventricular hypertrophy is associated with the dephosphorylation of Cx43 and its translocation from the intercalated disks to intracellular pools, suggesting accelerated gap junction degradation.
Sasano
Chieko
C
Department of Cardiovascular Research, Research Institute of Environmental Medicine, Nagoya University, Nagoya, Japan.
Honjo
Haruo
H
Takagishi
Yoshiko
Y
Uzzaman
Mahmud
M
Emdad
Luni
L
Shimizu
Atsuya
A
Murata
Yoshiharu
Y
Kamiya
Kaichiro
K
Kodama
Itsuo
I
D000818:Animals; D018031:Connexin 43 / Q000032:analysis* / Q000378:metabolism*; D004195:Disease Models, Animal; D004705:Endocytosis; D017629:Gap Junctions / Q000737:chemistry / Q000378:metabolism; D006976:Hypertension, Pulmonary / Q000150:complications*; D017380:Hypertrophy, Right Ventricular / Q000139:chemically induced / Q000473:pathology*; D032383:Myocytes, Cardiac / Q000737:chemistry / Q000473:pathology; D010766:Phosphorylation; D051381:Rats; D017208:Rats, Wistar; D014018:Tissue Distribution
D018031:Connexin 43