Rowlerson A, Scapolo P A, Mascarello F, Carpenè E, Veggetti A
J Muscle Res Cell Motil. 1985 Oct;6(5):601-40. doi: 10.1007/BF00711917.
Myosin isoforms and their distribution in the various fibre types of the lateral muscle of eight teleost fish (representing a wide range of taxonomic groups and lifestyles) were investigated electrophoretically, histochemically and immunohistochemically. Polyclonal antisera were raised against slow (red muscle) and fast (white muscle) myosins of the mullet, and used to stain sections of lateral muscle. Antisera specific for fast and slow myosin heavy chains only (anti-FHC and anti-SHC respectively) and for whole fast and slow myosins (anti-F and anti-S respectively) were obtained, and their specificity was confirmed by immunoblotting against electrophoretically separated myofibrillar proteins. The ATPase activity of myosin isoforms was examined histochemically using methods to demonstrate their acid- and alkali-lability and their Ca-Mg dependent actomyosin ATPase. As expected, the predominant myosin (and fibre) type in the red muscle showed an alkali-labile ATPase activity, reacted with the anti-S and anti-SHC sera (but not anti-F or anti-FHC) and contained two 'slow' light chains, whereas the predominant myosin (and fibre) type in the white muscle showed an alkali-stable ATPase activity, reacted with anti-F and anti-FHC sera (but not anti-S or anti-SHC) and contained three 'fast' light chains. However, superimposed upon this basic pattern were a number of variations, many of them species-related. On analysis by two-dimensional gel electrophoresis fish myosin light chains LC1s, LC2s and LC2f migrated like the corresponding light chains of mammalian myosins, but fish LC1f consistently had a more acidic pI value than mammalian LC1f. Fish LC3f varied markedly in Mr in a species-related manner: in some fish (e.g. eel and mullet) the Mr value of LC3f was less than that for the other two light chains (as in mammalian myosin), whereas in others it was similar to that of LC2f (e.g. cat-fish) or even greater (e.g. goldfish). Species differences were also seen in the relative intensity of LC1f and LC3f spots given by the fish fast myosins. In most of the fish examined the red muscle layer showed some micro-heterogeneity, containing (in addition to the typical slow fibres) small numbers of fibres with a histo- and immunohistochemical profile typical of white muscle (fast) fibres. However, other immunohistochemically distinct minority fibres were found in the red muscle of the goldfish. Three types of pink muscle were distinguished: a mosaic of immunohistochemically typical red and white fibres (e.g. grey mullet).(ABSTRACT TRUNCATED AT 400 WORDS)
对8种硬骨鱼（代表广泛的分类群和生活方式）侧肌不同纤维类型中的肌球蛋白异构体及其分布进行了电泳、组织化学和免疫组织化学研究。制备了针对鲻鱼慢肌（红肌）和快肌（白肌）肌球蛋白的多克隆抗血清，并用于对侧肌切片进行染色。获得了仅对快肌和慢肌球蛋白重链特异的抗血清（分别为抗FHC和抗SHC）以及对完整快肌和慢肌球蛋白特异的抗血清（分别为抗F和抗S），并通过对电泳分离的肌原纤维蛋白进行免疫印迹来确认其特异性。使用显示其酸稳定性和碱稳定性以及其钙 - 镁依赖性肌动球蛋白ATP酶的方法，通过组织化学检查肌球蛋白异构体的ATP酶活性。正如预期的那样，红肌中主要的肌球蛋白（和纤维）类型显示出碱不稳定的ATP酶活性，与抗S和抗SHC血清反应（但不与抗F或抗FHC反应），并含有两条“慢”轻链，而白肌中主要的肌球蛋白（和纤维）类型显示出碱稳定的ATP酶活性，与抗F和抗FHC血清反应（但不与抗S或抗SHC反应），并含有三条“快”轻链。然而，在这种基本模式之上存在许多变化，其中许多与物种有关。通过二维凝胶电泳分析，鱼类肌球蛋白轻链LC1s、LC2s和LC2f的迁移方式与哺乳动物肌球蛋白相应的轻链相似，但鱼类LC1f的pI值始终比哺乳动物LC1f更酸性。鱼类LC3f的分子量在物种相关的方式上有显著变化：在一些鱼类（如鳗鱼和鲻鱼）中，LC3f的分子量值小于其他两条轻链（如哺乳动物肌球蛋白），而在其他鱼类中，它与LC2f相似（如鲶鱼）甚至更大（如金鱼）。在鱼类快肌球蛋白给出的LC1f和LC3f斑点的相对强度上也观察到物种差异。在大多数被检查的鱼类中，红肌层表现出一些微异质性，除了典型的慢纤维外，还含有少量具有白肌（快）纤维典型组织化学和免疫组织化学特征的纤维。然而，在金鱼的红肌中发现了其他免疫组织化学上不同的少数纤维。区分出三种类型的粉色肌：免疫组织化学上典型的红纤维和白纤维的镶嵌（如灰鲻）。（摘要截于400字）
