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Eyestalk ablation has little effect on actin and myosin heavy chain gene expression in adult lobster skeletal muscles

机译:眼柄消融对成年龙虾骨骼肌肌动蛋白和肌球蛋白重链基因表达的影响很小

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The organization of skeletal muscles in decapod crustaceans is significantly altered during molting and development. Prior to molting, the claw muscles atrophy dramatically, facilitating their removal from the base of the claw. During development, lobster claw muscles exhibit fiber switching over several molt cycles. Such processes may be influenced by the secretion of steroid molting hormones, known collectively as ecdysteroids. To assay the effects of these hormones, we used eyestalk ablation to trigger an elevation of circulating ecdysteroids and then quantified myofibrillar mRNA levels with real-time PCR and myofibrillar protein levels by SDS-PAGE. Levels of myosin heavy chain (MHC) and actin proteins and the mRNA encoding them were largely unaffected by eyestalk ablation, but in muscles from intact animals, myofibrillar gene expression was modestly elevated in premolt and postmolt animals. In contrast, polyubiquitin mRNA was significantly elevated (about 2-fold) in claw muscles from eyestalk-ablated animals with elevated circulating ecdysteroids. Moreover, patterns of MHC and actin gene expression are significantly different among slow and fast claw muscles. Consistent with these patterns, the three muscle types differed in the relative amounts of myosin heavy chain and actin proteins. All three muscles also co-expressed fast and slow myosin isoforms, even in fibers that are generally regarded as exclusively fast or slow. These results are consistent with other recent data demonstrating co-expression of myosin isoforms in lobster muscles.
机译:在蜕皮和发育过程中,十足纲甲壳动物的骨骼肌组织发生了显着变化。蜕皮之前,爪状肌肉明显萎缩,有利于它们从爪基部移出。在发育过程中,龙虾爪肌肉在几个蜕皮周期中表现出纤维转换。此类过程可能会受到类固醇蜕皮激素(统称为蜕皮类固醇)的分泌的影响。为了测定这些激素的作用,我们使用了眼柄消融术来引发循环蜕皮激素的升高,然后通过实时PCR定量肌原纤维mRNA水平,并通过SDS-PAGE定量肌原纤维蛋白水平。肌球蛋白重链(MHC)和肌动蛋白蛋白以及编码它们的mRNA的水平在很大程度上不受眼球消融的影响,但是在完整动物的肌肉中,在蜕皮前和蜕皮后动物中,肌原纤维基因的表达适度升高。相比之下,患有眼睑切除的动物中循环蜕皮激素水平升高,其爪肌中的聚泛素mRNA显着升高(约2倍)。此外,MHC和肌动蛋白基因表达的模式在慢速和快速爪肌之间显着不同。与这些模式一致,这三种肌肉类型的肌球蛋白重链和肌动蛋白蛋白的相对量不同。即使在通常仅被视为快或慢的纤维中,所有三块肌肉也共表达快和慢肌球蛋白同工型。这些结果与其他最新数据一致,表明龙虾肌肉中肌球蛋白同种型的共表达。

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