Medler Scott, Brown Kitty J, Chang Ernest S, Mykles Donald L
Department of Biology, Colorado State University, Fort Collins, Colorado 80523, USA.
Biol Bull. 2005 Apr;208(2):127-37. doi: 10.2307/3593121.
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.
十足目甲壳动物的骨骼肌组织在蜕皮和发育过程中会发生显著变化。在蜕皮前,螯肌会大幅萎缩,便于从螯基部脱落。在发育过程中,龙虾的螯肌在几个蜕皮周期中会出现纤维转换。这些过程可能受类固醇蜕皮激素(统称为蜕皮甾类)分泌的影响。为了测定这些激素的作用,我们通过摘除眼柄来引发循环蜕皮甾类水平升高,然后用实时PCR定量肌原纤维mRNA水平,并用SDS-PAGE定量肌原纤维蛋白水平。肌球蛋白重链(MHC)和肌动蛋白的蛋白质水平及其编码的mRNA水平在很大程度上不受眼柄摘除的影响,但在完整动物的肌肉中,肌原纤维基因表达在蜕皮前和蜕皮后的动物中略有升高。相比之下,在循环蜕皮甾类水平升高的眼柄摘除动物的螯肌中,多聚泛素mRNA显著升高(约2倍)。此外,慢螯肌和快螯肌中MHC和肌动蛋白基因的表达模式有显著差异。与这些模式一致,三种肌肉类型的肌球蛋白重链和肌动蛋白的相对含量不同。即使在通常被认为是纯快肌或纯慢肌的纤维中,所有三种肌肉也共同表达快肌和慢肌肌球蛋白异构体。这些结果与最近其他表明龙虾肌肉中肌球蛋白异构体共同表达的数据一致。