IMMUNOLOCALIZATION OF TRIADIN, DHP RECEPTORS, AND RYANODINE RECEPTORS IN ADULT AND DEVELOPING SKELETAL-MUSCLE OF RATS
IMMUNOLOCALIZATION OF TRIADIN, DHP RECEPTORS, AND RYANODINE RECEPTORS IN ADULT AND DEVELOPING SKELETAL-MUSCLE OF RATS
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DOI：
10.1002/mus.880181104
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发表时间：
1995-11-01
期刊：
影响因子：
3.4
通讯作者：
FERGUSON, DG
中科院分区：
文献类型：
--
作者：
CARL, SL;FELIX, K;FERGUSON, DG
二氢吡啶受体(DHPR)和兰尼定受体(RyR)是骨骼肌纤维三联体连接的蛋白质。最近，从兔骨骼肌重肌浆网(SR)小泡中纯化出一种新的95 kDa蛋白质Triadin，我们用间接免疫金EM将Triadin定位于分离的三联体中SR的连接面。此外，我们还利用间接免疫荧光技术在成年和发育中的大鼠骨骼肌中定位了与DHPR和RyR相关的Triadin。在纵向定向的成年大鼠骨骼肌组织的双重免疫标记实验中，Triadin特异的抗体和RyR特异的抗体导致了特有的条纹染色模式。当对数字化激光扫描共聚焦显微镜图像进行计算机分析时，这些抗体产生的染色完全重叠。使用针对DHPR和RyR的抗体进行的双重染色实验中也得到了类似的结果，表明这三种蛋白都位于原位三联体中。通过对几个胎龄和出生后早期的骨骼肌组织进行双重标记，研究了这三个三联体蛋白的发育表达。Triadin、RyR和DHPR抗体的染色模式在整个发育过程中是重叠的，这表明从它们最早出现的时候起，这三种蛋白质就是三联体的组成部分。(C)1995 John Wiley&Sons，Inc.
The dihydropyridine receptors (DHPR) and the ryanodine receptors (RyR) are well-characterized proteins of the triad junctions of skeletal muscle fibers. Recently, a newly discovered 95-kDa protein, triadin, has been purified from rabbit skeletal muscle heavy sarcoplasmic reticulum (SR) vesicles, WE have used indirect immunogold EM to localize triadin to the junctional face of the SR in isolated triads. In addition, we have used indirect immunofluorescence to localize triadin in relation to the DHPR and the RyR in adult and developing rat skeletal muscle. In double immunolabeling experiments of longitudinally oriented adult rat skeletal muscle tissue, triadin-specific and RyR-specific antibodies resulted in a characteristic striated staining pattern. The staining arising from these antibodies completely overlapped when examined by computer analysis of digitized laser scanning confocal microscopy images. A similar result was obtained in double staining experiments using antibodies raised against the DHPR and the RyR suggesting that all three proteins are located in the triads in situ. The developmental expression of the three triad proteins was examined using double labeling of skeletal muscle tissue from several fetal and early postnatal ages. The staining patterns of triadin, RyR, and DHPR antibodies were overlapping throughout development, suggesting that from their earliest appearance the three proteins are components of the triads. (C) 1995 John Wiley & Sons, Inc.