Overexpression of phospholemman alters contractility and [Ca(2+)](i) transients in adult rat myocytes.
Previous studies showed increased phospholemman (PLM) mRNA after myocardial infarction (MI) in rats (Sehl PD, Tai JTN, Hillan KJ, Brown LA, Goddard A, Yang R, Jin H, and Lowe DG. Circulation 101: 1990-1999, 2000). We tested the hypothesis that, in normal adult rat cardiac myocytes, PLM overexpression alters contractile function and cytosolic Ca(2+) concentration ([Ca(2+)](i)) homeostasis in a manner similar to that observed in post-MI myocytes. Compared with myocytes infected by control adenovirus expressing green fluorescent protein (GFP) alone, Western blots indicated a 41% increase in PLM expression after 72 h (P < 0.001) but no changes in Na(+)/Ca(2+) exchanger, SERCA2, and calsequestrin levels in myocytes infected by adenovirus expressing GFP and PLM. At 5 mM extracellular [Ca(2+)] ([Ca(2+)](o)), maximal contraction amplitudes in PLM-overexpressed myocytes were 24% (P < 0.005) and [Ca(2+)](i) transient amplitudes were 18% (P < 0.05) lower than control myocytes. At 0.6 mM [Ca(2+)](o), however, contraction and [Ca(2+)](i) transient amplitudes were significantly (P < 0.05) higher in PLM-overexpressed than control myocytes (18% and 42%, respectively); at 1.8 mM [Ca(2+)](o), the differences in contraction and [Ca(2+)](i) transient amplitudes were narrowed. This pattern of contractile and [Ca(2+)](i) transient abnormalities in PLM-overexpressed myocytes mimics that observed in post-MI rat myocytes. We suggest that PLM overexpression observed in post-MI myocytes may partly account for contractile abnormalities by perturbing Ca(2+) fluxes during excitation-contraction.
Weis Center for Research, Geisinger Medical Center, Danville, PA 17822, USA.
Jianliang
J
Song
Xue-Qian
XQ
Zhang
Lois L
LL
Carl
Anwer
A
Qureshi
Lawrence I
LI
Rothblum
Joseph Y
JY
Cheung
D000257:Adenoviridae Infections / Q000503:physiopathology; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002304:Cardiac Pacing, Artificial; D002478:Cells, Cultured; D049452:Green Fluorescent Proteins; D007202:Indicators and Reagents; D007425:Intracellular Membranes / Q000378:metabolism*; D008164:Luminescent Proteins; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D009200:Myocardial Contraction / Q000502:physiology*; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D009994:Osmolar Concentration; D010750:Phosphoproteins / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
D007202:Indicators and Reagents; D008164:Luminescent Proteins; D008565:Membrane Proteins; D010750:Phosphoproteins; C069141:phospholemman; D049452:Green Fluorescent Proteins; D002118:Calcium