Phospholipase Cβ4 isozyme is expressed in human, rat, and murine heart left ventricles and in HL-1 cardiomyocytes
Phospholipase C-β (PLCβ) isozymes (PLCβ1 and PLCβ3) have been extensively characterized in cardiac tissue, but no data are available for the PLCβ4 isozyme. In this study, PLCβ(1–4) isozymes mRNA relative expression was studied by real-time PCR (RT-PCR) in human, rat, and murine left ventricle and the presence of PLCβ4 isozyme at the protein level was confirmed by Western blotting in all species studied. Confocal microscopy experiments carried out in HL-1 cardiomyocytes revealed a sarcoplasmic subcellular distribution of PLCβ4. Although there were unexpected significant interspecies differences in the PLCβ(1–4) mRNA expression, PLCβ4 mRNA was the main transcript expressed in all left ventricles studied. Thus, whereas in human and rat left ventricles PLCβ4 > PLCβ3 > PLCβ2 > PLCβ1 mRNA pattern of expression was found, in murine left ventricle the pattern of expression was different, i.e., PLCβ4 > PLCβ1 > PLCβ3 > PLCβ2. However, results obtained in mouse HL-1 cardiomyocytes showed PLCβ3 ≈ PLCβ4 > PLCβ1 > PLCβ2 pattern of mRNA expression indicating a probable cell type specific expression of the different PLCβ isozymes in cardiomyocytes. Finally, RT-PCR experiments showed a trend, even though not significant (P = 0.067), to increase PLCβ4 mRNA levels in HL-1 cardiomyocytes after angiotensin II treatment. These results demonstrate the presence of PLCβ4 in the heart and in HL-1 cardiomyocytes showing a different species-dependent pattern of expression of the PLCβ(1–4) transcripts. We discuss the relevance of these findings in relation to the development of cardiac hypertrophy.
KeywordsPhospholipase Cβ4 Human Rat Mouse Left ventricle RT-PCR Angiotensin II
The authors would like to thank Dr Eneko Urizar for his valuable advice at the time of this manuscript preparation. We would like to thank the cardiac surgeons from Policlinica Guipuzcoa for providing the human left ventricle sample biopsy. We thank to Dr. A. Aiastui for help in immunofluorescence technique; A. Pavón and A. Dorronsoro from Inbiomed for assistance with confocal microscopy This work was supported by the Universidad del País Vasco/Euskal Herriko Unibertsitatea (1/UPV 00079.252-E-15424/2003 to M.A.G.); and the Gobierno Vasco (GV2005111012 to M.A.G.). A.L. holds a fellowship from the Diputacion Foral from Gipuzkoa.
- 6.Filtz TM, Grubb DR, McLeod-Dryden TJ, Luo J, Woodcock EA (2009) Gq-inititated cardiomyocyte hypertrophy is mediated by phospholipase Cβ1b. FASEB J. doi: 10.1096/fj.09-133983
- 13.Strassheim D, Williams CL (2000) P2Y2 purinergic and M3 muscarinic acetylcholine receptors activate different phospholipase C-beta isoforms that are uniquely susceptible to protein kinase C-dependent phosphorylation and inactivation. J Biol Chem 275:39767–39772. doi: 10.1074/jbc.M007775200 CrossRefPubMedGoogle Scholar
- 15.Garro MA, López de Jesús M, Ruíz de Azúa I, Callado LF, Meana JJ, Sallés J (2001) Regulation of phospholipase Cβ activity by muscarinic acetylcholine and 5-HT2 receptors in crude and synaptosomal membranes from human cerebral cortex. Neuropharmacology 40:686–695. doi: 10.1016/S0028-3908(00)00206-9 CrossRefPubMedGoogle Scholar
- 20.John DY, Lee HH, Park D, Lee CW, Lee KH, Yoo OJ, Rhee SG (1993) Cloning, sequencing, purification and Gq-dependent activation of phospholipase C-β3. J Biol Chem 268:6654–6661Google Scholar
- 29.Bendall JK, Damy T, Ratajczak P, Loyer X, Monceau V, Marty I, Milliez P, Robidel E, Marotte F, Samuel JL, Heymes C (2004) Role of myocardial neuronal nitric oxide synthase-derived nitric oxide in beta-adrenergic hyporesponsiveness after myocardial infarction-induced heart failure in rat. Circulation 110:2368–2375. doi: 10.1161/01.CIR.0000145160.04084.AC CrossRefPubMedGoogle Scholar