Increased mechanical extraction of T-tubule/junctional SR from cardiomyopathic hamster heart.
We determined the contents of L-type calcium channels (LCC) and other membrane proteins in ventricular homogenates and microsomes prepared from hearts of 30- to 70-day-old Syrian cardiomyopathic (Bio 14.6) and normal hamsters. Quantitative immunoblot assay revealed that myopathic microsomes, as compared with normal controls, were enriched about twofold with the alpha 1-subunit of LCC, the ryanodine receptor calsequestrin, and Na(+)-K(+)-adenosinetriphosphatase (ATPase), whereas the contents of these proteins in ventricular homogenates were not different. In contrast, Na(+)-H+ antiporter and sarcoplasmic reticulum (SR) Ca(2+)-ATPase showed no difference in their contents in both homogenates and microsomes. Radioligand binding assay further showed no significant difference in the number of binding sites for [3H]prazosin, [125I]iodocyanopindolol, and [3H]saxitoxin between myopathic and normal microsomes. These result suggest that whereas membrane densities of LCC and the other proteins examined are not increased in myopathic cardiomyocytes, T-tubule/junctional SR membranes are more easily extracted from them by mechanical disruption. This, together with 1.5-fold higher yield of microsomal fractions from myopathic heart muscle, shows that abnormality exists in the mechanical property of cell membrane in the myopathic heart.
Tawada-Iwata
Y
Y
Department of Molecular Physiology, National Cardiovascular Center Research Institute, Osaka, Japan.
Imagawa
T
T
Yoshida
A
A
Takahashi
M
M
Nakamura
H
H
Shigekawa
M
M
D000818:Animals; D015220:Calcium Channels / Q000378:metabolism; D009202:Cardiomyopathies / Q000378:metabolism* / Q000473:pathology; D006224:Cricetinae; D006352:Heart Ventricles; D006652:Histological Techniques; D015151:Immunoblotting; D008024:Ligands; D008647:Mesocricetus; D008861:Microsomes / Q000378:metabolism / Q000648:ultrastructure; D009124:Muscle Proteins / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism* / Q000648:ultrastructure; D012016:Reference Values; D019837:Ryanodine Receptor Calcium Release Channel; D012519:Sarcoplasmic Reticulum / Q000378:metabolism*
D015220:Calcium Channels; D008024:Ligands; D009124:Muscle Proteins; D019837:Ryanodine Receptor Calcium Release Channel