scholarly journals Molecular cloning and mRNA expression analysis of carp embryonic, slow and cardiac myosin heavy chain isoforms

2006 ◽  
Vol 209 (1) ◽  
pp. 188-198 ◽  
Author(s):  
Y. Nihei
Keyword(s):  
Mrna Expression ◽  
Myosin Heavy Chain ◽  
Molecular Cloning ◽  
Expression Analysis ◽  
Heavy Chain ◽  
Myosin Heavy Chain Isoforms ◽  
Cardiac Myosin ◽  
Mrna Expression Analysis

scholarly journals Electrophoretic separation and quantitation of cardiac myosin heavy chain isoforms in eight mammalian species

1998 ◽  
Vol 274 (3) ◽  
pp. H1048-H1053 ◽  
Author(s):  
Peter J. Reiser ◽  
William O. Kline
Keyword(s):  
Skeletal Muscle ◽  
Sample Preparation ◽  
Myosin Heavy Chain ◽  
Heavy Chain ◽  
Mammalian Species ◽  
Myosin Heavy Chain Isoforms ◽  
Glycerol Concentration ◽  
Cardiac Myosin ◽  
Homogenization Step

A protocol for sample preparation and gel electrophoresis is described that reliably results in the separation of the α- and β-isoforms of cardiac myosin heavy chain (MHC-α and MHC-β) in eight mammalian species. The protocol is based on a simple, nongradient denaturing gel. The magnitude of separation of MHC-α and MHC-β 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-α and MHC-β in myocardial samples from different species. The effect of sample load on MHC-α and MHC-β 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.

Molecular cloning and mRNA expression analysis of sheep MYL3 and MYL4 genes

Gene ◽  
2016 ◽  
Vol 577 (2) ◽  
pp. 209-214 ◽  
Author(s):  
Chunlan Zhang ◽  
Jianmin Wang ◽  
Guizhi Wang ◽  
Zhibin Ji ◽  
Lei Hou ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Molecular Cloning ◽  
Expression Analysis ◽  
Mrna Expression Analysis

Molecular cloning, characterization and mRNA expression analysis of a novel selenoprotein: avian selenoprotein W from chicken

2010 ◽  
Vol 38 (6) ◽  
pp. 4015-4022 ◽  
Author(s):  
Jin-Long Li ◽  
Hong-Feng Ruan ◽  
Hui-Xin Li ◽  
Shu Li ◽  
Shi-Wen Xu ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Molecular Cloning ◽  
Expression Analysis ◽  
Selenoprotein W ◽  
Mrna Expression Analysis

scholarly journals Regulatory light chain phosphorylation augments length-dependent contraction in PTU-treated rats

2018 ◽  
Vol 151 (1) ◽  
pp. 66-76 ◽  
Author(s):  
Jason J. Breithaupt ◽  
Hannah C. Pulcastro ◽  
Peter O. Awinda ◽  
David C. DeWitt ◽  
Bertrand C.W. Tanner
Keyword(s):  
Myosin Heavy Chain ◽  
Heavy Chain ◽  
Light Chain ◽  
Myosin Heavy Chain Isoforms ◽  
Regulatory Light Chain ◽  
Cardiac Myosin ◽  
Myosin Heavy Chain Isoform ◽  
Cross Bridge ◽  
Detachment Rate ◽  
Length Dependent Activation

Force production by actin–myosin cross-bridges in cardiac muscle is regulated by thin-filament proteins and sarcomere length (SL) throughout the heartbeat. Prior work has shown that myosin regulatory light chain (RLC), which binds to the neck of myosin heavy chain, increases cardiac contractility when phosphorylated. We recently showed that cross-bridge kinetics slow with increasing SLs, and that RLC phosphorylation amplifies this effect, using skinned rat myocardial strips predominantly composed of the faster α-cardiac myosin heavy chain isoform. In the present study, to assess how RLC phosphorylation influences length-dependent myosin function as myosin motor speed varies, we used a propylthiouracil (PTU) diet to induce >95% expression of the slower β-myosin heavy chain isoform in rat cardiac ventricles. We measured the effect of RLC phosphorylation on Ca2+-activated isometric contraction and myosin cross-bridge kinetics (via stochastic length perturbation analysis) in skinned rat papillary muscle strips at 1.9- and 2.2-µm SL. Maximum tension and Ca2+ sensitivity increased with SL, and RLC phosphorylation augmented this response at 2.2-µm SL. Subtle increases in viscoelastic myocardial stiffness occurred with RLC phosphorylation at 2.2-µm SL, but not at 1.9-µm SL, thereby suggesting that RLC phosphorylation increases β-myosin heavy chain binding or stiffness at longer SLs. The cross-bridge detachment rate slowed as SL increased, providing a potential mechanism for prolonged cross-bridge attachment to augment length-dependent activation of contraction at longer SLs. Length-dependent slowing of β-myosin heavy chain detachment rate was not affected by RLC phosphorylation. Together with our previous studies, these data suggest that both α- and β-myosin heavy chain isoforms show a length-dependent activation response and prolonged myosin attachment as SL increases in rat myocardial strips, and that RLC phosphorylation augments length-dependent activation at longer SLs. In comparing cardiac isoforms, however, we found that β-myosin heavy chain consistently showed greater length-dependent sensitivity than α-myosin heavy chain. Our work suggests that RLC phosphorylation is a vital contributor to the regulation of myocardial contractility in both cardiac myosin heavy chain isoforms.

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