| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Department of Oral Biology, The Ohio State University, Columbus, Ohio 43210-1241
A protocol for
sample preparation and gel electrophoresis is described that reliably
results in the separation of the 
- and 
-isoforms of cardiac
myosin heavy chain (MHC- and MHC-) in eight mammalian species.
The protocol is based on a simple, nongradient denaturing gel. The
magnitude of separation of MHC- and MHC- achieved with this
protocol is sufficient for quantitative determination of the relative
amounts of these two isoforms in mouse, rat, guinea pig, rabbit,
canine, pig, baboon, and human myocardial samples. The sensitivity of
the protocol is sufficient for the detection of MHC isoforms in samples
at least as small as 1 µg. The glycerol concentration in the
separating gel is an important factor for successfully separating
MHC- and MHC- in myocardial samples from different species. The
effect of sample load on MHC- and MHC- band resolution is
illustrated. The results also indicate that inclusion of a
homogenization step during sample preparation increases the amount of
MHC detected on the gel for cardiac samples to a much greater extent
than for skeletal muscle samples. Although the protocol described in
this study is excellent for analyzing cardiac samples, it should be
noted that the same protocol is not optimal for separating MHC isoforms
expressed in skeletal muscle, as is illustrated.
isoenzymes; sodium dodecyl sulfate-polyacrylamide gel electrophoresis; contractile proteins; myocardium; heart
This article has been cited by other articles:
|
|
 |
|
  B. M. Palmer, Y. Wang, P. Teekakirikul, J. T. Hinson, D. Fatkin, S. Strouse, P. VanBuren, C. E. Seidman, J. G. Seidman, and D. W. Maughan Myofilament mechanical performance is enhanced by R403Q myosin in mouse myocardium independent of sex Am J Physiol Heart Circ Physiol, April 1, 2008; 294(4): H1939 - H1947. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Guellich, T. Damy, Y. Lecarpentier, M. Conti, V. Claes, J.-L. Samuel, J. Quillard, J.-L. Hebert, T. Pineau, and C. Coirault Role of oxidative stress in cardiac dysfunction of PPAR{alpha}-/- mice Am J Physiol Heart Circ Physiol, July 1, 2007; 293(1): H93 - H102. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Bergrin, S. Bicer, C. A. Lucas, and P. J. Reiser Three-dimensional compartmentalization of myosin heavy chain and myosin light chain isoforms in dog thyroarytenoid muscle Am J Physiol Cell Physiol, May 1, 2006; 290(5): C1446 - C1458. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. C. Blunt, Y. Chen, J. D. Potter, and P. A. Hofmann Modest actomyosin energy conservation increases myocardial postischemic function Am J Physiol Heart Circ Physiol, March 1, 2005; 288(3): H1088 - H1096. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. L. M. Rundell, V. Manaves, A. F. Martin, and P. P. de Tombe Impact of {beta}-myosin heavy chain isoform expression on cross-bridge cycling kinetics Am J Physiol Heart Circ Physiol, February 1, 2005; 288(2): H896 - H903. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Hashimoto, N. Kambara, R. Nohara, M. Yazawa, and S. Taguchi Expression of MHC-{beta} and MCT1 in cardiac muscle after exercise training in myocardial-infarcted rats J Appl Physiol, September 1, 2004; 97(3): 843 - 851. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. A. Carnes, T. P. Geisbuhler, and P. J. Reiser Age-dependent changes in contraction and regional myocardial myosin heavy chain isoform expression in rats J Appl Physiol, July 1, 2004; 97(1): 446 - 453. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Iemitsu, T. Miyauchi, S. Maeda, T. Tanabe, M. Takanashi, M. Matsuda, and I. Yamaguchi Exercise training improves cardiac function-related gene levels through thyroid hormone receptor signaling in aged rats Am J Physiol Heart Circ Physiol, May 1, 2004; 286(5): H1696 - H1705. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Zhong, P. J Reiser, and M. A. Matlib Gender differences in myosin heavy chain-{beta} and phosphorylated phospholamban in diabetic rat hearts Am J Physiol Heart Circ Physiol, December 1, 2003; 285(6): H2688 - H2693. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. R. Alpert, C. Brosseau, A. Federico, M. Krenz, J. Robbins, and D. M. Warshaw Molecular mechanics of mouse cardiac myosin isoforms Am J Physiol Heart Circ Physiol, October 1, 2002; 283(4): H1446 - H1454. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. J. Mihm, F. Yu, C. A. Carnes, P. J. Reiser, P. M. McCarthy, D. R. Van Wagoner, and J. A. Bauer Impaired Myofibrillar Energetics and Oxidative Injury During Human Atrial Fibrillation Circulation, July 10, 2001; 104(2): 174 - 180. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. L. Clanton, V. P. Wright, P. J. Reiser, P. F. Klawitter, and N. R. Prabhakar Physiological and Genomic Consequences of Intermittent Hypoxia: Selected Contribution: Improved anoxic tolerance in rat diaphragm following intermittent hypoxia J Appl Physiol, June 1, 2001; 90(6): 2508 - 2513. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. J. Reiser, M. A. Portman, X.-H. Ning, and C. S. Moravec Human cardiac myosin heavy chain isoforms in fetal and failing adult atria and ventricles Am J Physiol Heart Circ Physiol, April 1, 2001; 280(4): H1814 - H1820. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Wattanapermpool and P. J. Reiser Differential effects of ovariectomy on calcium activation of cardiac and soleus myofilaments Am J Physiol Heart Circ Physiol, August 1, 1999; 277(2): H467 - H473. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H.-J. Wang, Y.-C. Zhu, and T. Yao Effects of all-trans retinoic acid on angiotensin II-induced myocyte hypertrophy J Appl Physiol, May 1, 2002; 92(5): 2162 - 2168. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| Visit Other APS Journals Online |
