[Integration between M3 muscarinic acetylcholine receptor and connexin 43 as antiarrhythmic targets in rat ventricular myocardium].
OBJECTIVE
To optimize the method of investigating structural integration between proteins and study the integration between arrhythmia related proteins in molecular level.


METHODS
Immunostaining the normal ventricular myocytes was used to observe the distribution of connexin 43 and muscarinic acetylcholine receptor (mAChR). The five mAChR subtypes were precipitated using immunoprecipitation. Then, SDS-PAGE and Western blotting with the anti-connexin 43 antibody were performed to observe whether they were structurally integrated. Further, different concentrations of detergent were used to observe whether this relationship could be broken.


RESULTS
The five subtypes of mAChR existed in the cardiac myocyte of the rat, and all the five mAChR subtypes combined with connexin 43. In the normal rat ventricular myocyte membrane, connexin 43 and M3 receptor are co-located. When adding certain concentration of detergent to the membrane protein, the integration between M3 receptor and connexin 43 was broken, and the phosphorylated form of connexin 43 integrated with M3 receptor.


CONCLUSIONS
The results indicated that the structural integration between mAChR and phosphorylation of connexin 43 existed in rat ventricular myocardium, and this integration could be broken by certain concentration of detergent.
Zhang
Yong
Y
Department of Pharmacology, Bio-pharmaceutical Key Laboratory of Heilongjiang Province-Incubator of State Key Laboratory, Harbin Medical University, Harbin 150086, China.
Yue
Peng
P
Xiao
Jing
J
Yu
Hai-Yan
HY
Pan
Zhen-Wei
ZW
Lin
Dao-Hong
DH
Lü
Yan-Jie
YJ
Yang
Bao-Feng
BF
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D018031:Connexin 43 / Q000378:metabolism*; D006352:Heart Ventricles; D047468:Immunoprecipitation; D008297:Male; D018613:Microscopy, Confocal; D032383:Myocytes, Cardiac / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D051381:Rats; D017208:Rats, Wistar; D043587:Receptor, Muscarinic M3 / Q000378:metabolism*; D011976:Receptors, Muscarinic / Q000378:metabolism*; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology
D018031:Connexin 43; D043587:Receptor, Muscarinic M3; D011976:Receptors, Muscarinic; D012967:Sodium Dodecyl Sulfate