Whitmer J T, Kumar P, Solaro R J
Department of Pediatrics, Children's Hospital Medical Center, Cincinnati, OH 45229.
Circ Res. 1988 Jan;62(1):81-5. doi: 10.1161/01.res.62.1.81.
Calcium uptake was measured in homogenates and microsomal preparations enriched in sarcoplasmic reticulum vesicles isolated from hearts of hypertrophic (BIO 14.6) and dilated (BIO 53.58) cardiomyopathic as well as control (F1B) Syrian hamsters at 3, 7, 9, and 11 months of age. Calcium uptake studies were done using the Millipore filtration technique under conditions known to restrict transport to the sarcoplasmic reticulum. Steady-state calcium uptake capacity was used as a measure of the relative amounts of sarcoplasmic reticulum in homogenates prepared from individual hearts. At 3 months of age, there were no differences in calcium uptake in homogenates from control or myopathic hearts. However, by 9 months, although calcium uptake of homogenates from control and hypertrophic hearts was the same, calcium uptake by homogenates from dilated hearts was significantly depressed both in initial rate and capacity. Similar trends were seen in the microsomal vesicle preparations, but the decrease in calcium uptake in the dilated hearts was significantly lower by 3 months of age. The catalytic activity of the sarcoplasmic reticulum transport enzyme was estimated from the ratio of velocity to capacity, which provides a measure of the fractional rate of filling of the sarcoplasmic reticulum with calcium. The velocity-to-capacity ratios were not different at any of the ages in both the homogenate and microsomal preparations. The results of this study demonstrate that a major defect in the dilated cardiomyopathy may be due to a decrease in the volume or number of sarcoplasmic reticulum calcium transport sites rather than a decrease in specific activity of the transport enzyme.
在3、7、9和11月龄的肥厚型(BIO 14.6)、扩张型(BIO 53.58)心肌病叙利亚仓鼠以及对照(F1B)叙利亚仓鼠的心脏中,分离出富含肌浆网囊泡的匀浆和微粒体制剂,测定其中的钙摄取量。钙摄取研究采用密理博过滤技术,在已知限制钙转运至肌浆网的条件下进行。稳态钙摄取能力用作衡量从各个心脏制备的匀浆中肌浆网相对含量的指标。3月龄时,对照或心肌病心脏匀浆中的钙摄取量没有差异。然而,到9月龄时,尽管对照和肥厚型心脏匀浆的钙摄取量相同,但扩张型心脏匀浆的钙摄取在初始速率和摄取能力方面均显著降低。微粒体囊泡制剂中也观察到类似趋势,但扩张型心脏钙摄取量的降低在3月龄时明显更低。肌浆网转运酶的催化活性通过速度与摄取能力的比值来估算,该比值可衡量肌浆网被钙填充的分数速率。在匀浆和微粒体制剂中,任何年龄的速度与摄取能力比值均无差异。本研究结果表明,扩张型心肌病的一个主要缺陷可能是由于肌浆网钙转运位点的数量或体积减少,而非转运酶的比活性降低。