An alpha-cardiac myosin heavy chain gene mutation impairs contraction and relaxation function of cardiac myocytes

Abstract

Left Ventricular (LV) myocytes were isolated from 15-wk-old male mice bearing the Arg403 --> Gln alpha-cardiac myosin heavy chain missense mutation (alpha-MHC403/+), a model of familial hypertrophic cardiomyopathy. LV myocytes were classified morphologically: type I, rod shaped with parallel myofibrils; type II, irregularly shaped, shorter and wider than wild-type (WT) control cells, with parallel myofibrils; and type III, irregularly shaped with disoriented myofibrils. Compared with WT myocytes, alpha-MHC403/+ myocytes had fewer type I cells (WT = 74 +/- 3%, alpha-MHC403/+ = 41 +/- 4%, P < 0.01) and more type III cells (WT= 12 +/- 3%, alpha-MHC403/+ = 49 +/- 7%, P < 0.01). In situ histology also demonstrated marked myofibrillar disarray in the alpha-MHC403/+ hearts. With the use of video edge detection, myocytes were paced at 1 Hz (37 degrees C) to determine the effects of the mutation on myocyte function. End-diastolic length was reduced in mutant myocytes, but fractional shortening (% contraction) and sarcomere length were not. Velocity of contraction (-dL/dtmax) was depressed in mutant cells, but more in type II and III cells (-31%) than in type I cells (-18%). Velocity of relaxation (+dL/dt) was also depressed more in type II and III cells (-38%) than in type I cells (-16%). Using fura 2 dye with intracellular Ca2+ transients, we demonstrated that in alpha-MHC403/+ myocytes, the amplitude of the Ca2+ signal during contraction was unchanged but that the time required for decay of the signal to decrease 70% from its maximum was delayed significantly (WT = 159 +/- 8 ms; alpha-MHC403/+ = 217 +/- 14 ms, P < 0.01). Sarco(endo)plasmic reticulum Ca2+-ATPase mRNA levels in alpha-MHC403/+ and WT mice were similar. These data indicate that the altered cardiac dysfunction of alpha-MHC403/+ myocytes is directly due to defective myocyte function rather than to secondary changes in global cardiac function and/or loading conditions.