Rearrangement of mRNAs for costamere proteins during costamere development in cultured skeletal muscle from chicken.
Rearrangement of mRNAs for costamere proteins during costamere development in cultured skeletal muscle from chicken.
复制标题
培养鸡骨骼肌肋骨发育过程中肋骨蛋白 mRNA 的重排。
作者：
E. Morris;A. Fulton
成熟的骨骼肌原纤维被中肋、后黏着斑蛋白的肋、黏着斑蛋白、中间丝和将肌原纤维连接到细胞外基质的其他蛋白质包围。最近已被证明是由肌原纤维产生的力被横向转导到细胞外基质中的位点。我们观察了培养的骨骼肌中的中肋骨的发育，从胚胎鸡腿中提取的细胞在微团培养中生长。我们用免疫荧光和原位杂交检测蛋白质和mRNA。抗体和探针信号通过激光扫描共聚焦显微镜成像。在融合后大约第12天，首先在与肌原纤维的Z线对齐的条纹中检测到波形蛋白的抗体;此后不久，在相同的条纹中也检测到波形蛋白mRNA的探针。光学切片表明波形蛋白mRNA和蛋白质非常接近，相距不超过0.1 mm，可能直接接触。波形蛋白抗体仅在自发抽搐的细胞中检测到条纹。接着，在融合后约17天，在相同周期性的条带中检测针对结蛋白和黏着斑蛋白蛋白和mRNA的抗体和探针。在发育的早期，在病灶接触处检测到白蛋白（而不是mRNA）。荧光渗出、RNase H敏感性、无探针杂交和与肌原纤维结合的对照均得到适当的结果。甘油醛-3-磷酸脱氢酶（一种糖酵解酶）的探针弥漫染色，与肌原纤维无关。这些结果表明，组成的costamere到达的结构在一个定义的序列，和mRNA组织是一个显着的，精确的和时间控制的方面costamere的发展。这些结果可能具有更广泛的影响。在这些细胞中，一些mRNA在空间上以亚微米的精度定位，并且随着时间的推移而变化。特定的mRNA在这种定位的时间和地点上不同。这意味着细胞结构为这种定位提供了物理线索，而mRNA包含的信息以特定的方式与这些线索相互作用。如果在其他体细胞中发现mRNA定位的这种精确性，它可能对细胞产生和维持细胞结构的方式产生重大影响。
Mature skeletal myofibrils are surrounded by costameres, ribs of metavinculin, vinculin, intermediate filaments, and other proteins that connect the myofibril to the extracellular matrix. Costameres have recently been shown to be the sites at which the forces generated by the myofibril are transduced laterally into the extracellular matrix. We observed costameres developing in cultured skeletal muscles, grown in micromass culture from cells taken from embryonic chicken leg. We detected proteins by immunofluorescence and mRNA by in situ hybridization. Antibody and probe signals were imaged by laser scanning confocal microscopy. Antibody to vimentin protein is first detected in stripes in register with the Z line of the myofibril, at approximately day 12 after fusion; soon thereafter probe to vimentin mRNA is also detected in the same stripes. Optical sections indicate that vimentin mRNA and protein are very close, no more than 0.1 mm apart and possibly in immediate contact. Antibody to vimentin is detected in stripes only in cells that twitch spontaneously. Antibodies and probes to desmin and vinculin protein and mRNA are next detected in stripes of the same periodicity, at approximately day 17 after fusion. Vinculin protein (but not mRNA) is detected at focal contacts much earlier in development. Controls for bleed through of fluorescence, RNase H sensitivity, hybridization without probe, and binding to the myofibril all gave appropriate results. Probes to glyceraldehyde-3-phosphate dehydrogenase, a glycolytic enzyme, stained diffusely and did not associate with the myofibril. These results show that components of the costamere arrive at the structure in a defined sequence, and that mRNA organization is a conspicuous, precise and temporally controlled aspect of costamere development. These results may have wider implications. In these cells, some mRNAs are positioned with submicrometer precision in space and differentially over time. Particular mRNAs differ in the time and place of such positioning. This implies both that cellular structures provide physical cues for such positioning and that mRNA contains information that interacts with such cues in a message-specific manner. If such precision in mRNA location is found in other somatic cells, it could have significant implications for the ways in which cells generate and maintain cellular structures.
中等大小的丝：肌源性和神经源性谱系细胞的合成和分布的变化。
DOI：
10.1101/sqb.1982.046.01.033
发表时间：
1982
期刊：
Cold Spring Harbor symposia on quantitative biology
影响因子：
--
作者：
Holtzer,H;Bennett,GS;Tapscott,SJ;Croop,JM;Toyama,Y
通讯作者：
Toyama,Y
营养不良鸡肌肉中慢肌球蛋白的分布。
影响因子：
--
作者：
Bandman,E
通讯作者：
Bandman,E