A novel contractile phenotype with cardiac transgenic expression of the human P2X4 receptor.
The P2X4 receptor is a newly identified receptor expressed in the heart cell. Its function was elucidated with cardiac transgenic (TG) expression of the receptor by using the myocardium-specific a-myosin heavy chain promoter. The presence of the transgene was determined by polymerase chain reaction by using primers specific to the receptor and the vector linker region, by Southern blotting of the genomic DNA, and by immunoblotting and immunohistochemistry of both isolated cardiac myocytes and intact hearts. In intact heart study, the P2X4 receptor TG mouse exhibited significantly elevated basal cardiac contractility with greater rates of contraction and relaxation, left ventricular developed pressure, and cardiac output compared with nontransgenic (NTG) animals but showed no evidence of hypertrophy or heart failure. The TG heart also showed a greater increase of cardiac contractility in response to the P2X receptor agonist 2-methylthioATP, consistent with overexpression of a functional P2X4 receptor with consequent increase in the receptor-mediated response. In isolated cardiac cell study, the TG heart cell showed a similar level of basal contraction amplitude as the NTG heart cell while exhibiting a threefold greater increase in contractility during stimulation by 2-methylthioATP. Thus, an increased responsiveness of the overexpressed P2X4 receptor to endogenous ATP is responsible for the enhanced basal cardiac performance in the intact TG heart. The sustained enhanced contractile function with no associated heart pathology in the P2X4 receptor TG mouse suggests a novel physiologic role of the P2X4 receptor, that of stimulating the cardiac contractility.
Hu
B
B
Department of Medicine, Cardiovascular Division, University of Pennsylvania Medical Center, Philadelphia, Pennsylvania 19104, USA.
Mei
Q B
QB
Yao
X J
XJ
Smith
E
E
Barry
W H
WH
Liang
B T
BT
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D002302:Cardiac Output / Q000187:drug effects; D006321:Heart / Q000187:drug effects / Q000502:physiology; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic; D008954:Models, Biological; D009200:Myocardial Contraction / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D009206:Myocardium / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D018995:Myosin Heavy Chains / Q000235:genetics; D010641:Phenotype; D011401:Promoter Regions, Genetic / Q000235:genetics; D018048:Receptors, Purinergic P2 / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D058848:Receptors, Purinergic P2X4; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology; D013873:Thionucleotides / Q000494:pharmacology
C548226:P2RX4 protein, human; C548227:P2rx4 protein, mouse; D018048:Receptors, Purinergic P2; D058848:Receptors, Purinergic P2X4; D011993:Recombinant Fusion Proteins; D013873:Thionucleotides; D000255:Adenosine Triphosphate; D018995:Myosin Heavy Chains; C008501:2-methylthio-ATP