Department of Biology, Colorado State University, Fort Collins, CO 80523, USA.
Comp Biochem Physiol A Mol Integr Physiol. 2010 Dec;157(4):328-37. doi: 10.1016/j.cbpa.2010.07.024. Epub 2010 Aug 7.
A cDNA encoding a myostatin (Mstn)-like gene from an astacuran crustacean, Homarus americanus, was cloned and characterized. Mstn inhibits skeletal muscle growth in vertebrates and may play a role in crustacean muscle as a suppressor of protein synthesis. Sequence analysis and three-dimensional modeling of the Ha-Mstn protein predicted a high degree of conservation with vertebrate and other invertebrate myostatins. Qualitative polymerase chain reaction (PCR) demonstrated ubiquitous expression of transcript in all tissues, including skeletal muscles. Quantitative PCR analysis was used to determine the effects of natural molting and eyestalk ablation (ESA) on Ha-Mstn expression in the cutter claw (CT) and crusher claw (CR) closer muscles and deep abdominal (DA) muscle. In intermolt lobsters, the Ha-Mstn mRNA level in the DA muscle was significantly lower than the mRNA levels in the CT and CR muscles. Spontaneous molting decreased Ha-Mstn mRNA during premolt, with the CR muscle, which is composed of slow-twitch (S₁) fibers, responding preferentially (82% decrease) to the atrophic signal compared to fast fibers in CT (51% decrease) and DA (69% decrease) muscles. However, acute increases in circulating ecdysteroids caused by ESA had no effect on Ha-Mstn mRNA levels in the three muscles. These data indicate that the transcription of Ha-Mstn is differentially regulated during the natural molt cycle and it is an important regulator of protein turnover in molt-induced claw muscle atrophy.
从美洲螯龙虾(Homarus americanus)中克隆并鉴定了一种肌抑素(Mstn)样基因的 cDNA。Mstn 在脊椎动物中抑制骨骼肌生长,并且可能在甲壳动物肌肉中作为蛋白质合成的抑制剂发挥作用。Ha-Mstn 蛋白的序列分析和三维建模预测与脊椎动物和其他无脊椎动物的 Mstn 具有高度保守性。定性聚合酶链反应(PCR)证明了包括骨骼肌在内的所有组织中转录本的普遍表达。定量 PCR 分析用于确定天然蜕皮和眼柄切除(ESA)对切割爪(CT)和粉碎爪(CR)闭合肌和深腹肌(DA)中 Ha-Mstn 表达的影响。在蜕皮间期的龙虾中,DA 肌肉中的 Ha-Mstn mRNA 水平明显低于 CT 和 CR 肌肉中的 mRNA 水平。自发蜕皮使 CR 肌肉在蜕皮前期的 Ha-Mstn mRNA 减少,而 CR 肌肉由慢肌纤维(S₁)组成,与 CT(51%减少)和 DA(69%减少)肌肉中的快纤维相比,对萎缩信号的反应更为优先(82%减少)。然而,ESA 引起的循环蜕皮甾酮的急性增加对三种肌肉中的 Ha-Mstn mRNA 水平没有影响。这些数据表明,Ha-Mstn 的转录在自然蜕皮周期中受到差异调节,并且是蜕皮诱导的爪肌肉萎缩中蛋白质周转的重要调节剂。