Faculty of Life Sciences, The University of Manchester, Core Technology Facility 46 Grafton Street, Manchester, M13 9NT, UK.
J Comp Physiol B. 2009 Nov;179(8):961-9. doi: 10.1007/s00360-009-0377-x. Epub 2009 Jun 21.
In cardiomyocytes, ryanodine receptors (RYRs) mediate Ca(2+)-induced Ca(2+)-release (CICR) from the sarcoplasmic reticulum (SR) during excitation-contraction (e-c) coupling. In rainbow trout heart, the relative importance of CICR increases with cold-acclimation. Thus, the aim of this study was to investigate the effect of temperature acclimation (4, 11 and 18 degrees C) on RYR intracellular localization and expression density. We used immunocytochemistry to assess intracellular localization in ventricular myocytes and Western blotting to assess RYR expression in both atrial and ventricular tissue. In ventricular myocytes, RYRs were localized peripherally in transverse bands aligning with sarcomeric m-lines and centrally around mitochondria and the nucleus. Localization did not change with temperature acclimation. RYR expression was also unaffected by temperature acclimation. The localization of RYRs at the m-line is similar to neonatal mammalian cardiomyocytes. We suggest this positioning is indicative of myocytes which rely predominantly on transsarcolemmal Ca(2+)-influx, rather than CICR, during e-c coupling.
在心肌细胞中,兰尼碱受体(RYR)在兴奋-收缩(e-c)偶联期间介导肌浆网(SR)中的 Ca(2+)诱导的 Ca(2+)释放(CICR)。在虹鳟鱼心脏中,CICR 的相对重要性随着冷适应而增加。因此,本研究的目的是研究温度适应(4、11 和 18°C)对 RYR 细胞内定位和表达密度的影响。我们使用免疫细胞化学评估心室肌细胞中的细胞内定位,并用 Western blot 评估心房和心室组织中的 RYR 表达。在心室肌细胞中,RYR 定位于与肌节 m 线对齐的横带上,以及线粒体和核周围的中心位置。定位不受温度适应的影响。RYR 的表达也不受温度适应的影响。RYR 在 m 线上的定位与新生哺乳动物心肌细胞相似。我们认为这种定位表明心肌细胞在 e-c 偶联期间主要依赖跨肌细胞膜的 Ca(2+)内流,而不是 CICR。