HILL D K
J Cell Biol. 1964 Mar;20(3):435-58. doi: 10.1083/jcb.20.3.435.
The location of tritiated adenine nucleotide was studied by autoradiography in toad muscle which had been fixed in an acetic acid/ethanol mixture, with lead acetate present to act as the nucleotide precipitant. The muscles were embedded in Araldite. The autoradiographs were examined in both the light microscope and the electron microscope. In confirmation of earlier work, it was found that at least 50 per cent of the adenine nucleotide is concentrated in the I band, near the A-I boundary, and evidence is now available to suggest that this fraction is located in the interfibrillar spaces rather than in the substance of the fibrils. Electron micrographs of unstained sections failed to show any structural features at the site in question, though the triads were made visible by the lead. However, when sections were stained with an alkaline solution containing lead, or with uranyl acetate, "vesicles" were revealed at the appropriate site and it is thought that these may be elements of a transverse network (reticulum) of tubules containing the adenine nucleotide. The location of phosphocreatine could not be investigated because this substance was lost from the muscle during the preparative procedure.
利用放射自显影技术,在蟾蜍肌肉中研究了氚标记腺嘌呤核苷酸的定位情况。蟾蜍肌肉用乙酸/乙醇混合液固定,加入醋酸铅作为核苷酸沉淀剂。肌肉包埋于环氧树脂中。分别在光学显微镜和电子显微镜下观察放射自显影片。与早期研究结果一致,发现至少50%的腺嘌呤核苷酸集中在I带,靠近A-I边界,目前有证据表明这部分核苷酸位于肌原纤维间的间隙中,而非肌原纤维物质内。未染色切片的电子显微照片未显示该部位有任何结构特征,不过铅使三联管可见。然而,当切片用含铅碱性溶液或醋酸铀染色时,在相应部位发现了“囊泡”,据认为这些可能是含有腺嘌呤核苷酸的横向小管网络(网状结构)的组成部分。由于在制备过程中肌肉中的磷酸肌酸丢失,因此无法对其定位进行研究。