硬骨鱼(Poecilia latipinna)再生尾鳍中多肽的时间表达模式——以krt15和myl-1为重点

Hiral Murawala, Isha Ranadive, Sonam Patel, S. Balakrishnan
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引用次数: 6

摘要

在许多脊椎动物中,硬骨鱼的尾鳍已经成为理解再生机制的一个很好的模型。到目前为止,我们已经清楚地认识到,不同的调控信号需要适当的协调,以促进再生的连续阶段的进展。因此,在目前的研究中,研究的重点是鱿鱼尾鳍再生的蛋白表达谱。在不同的再生阶段观察到蛋白质的转换模式和表达水平的变化。二维凝胶电泳数据显示,与静息期相比,伤口上皮、囊胚和分化期的蛋白表达存在差异。基于肽表达谱的计算分析和随后的测序,我们在转录水平上进一步研究了角蛋白I型细胞骨架15和肌球蛋白轻链-1两种肽。角蛋白-15在创面上皮阶段表达上调,而在破胚和再生分化阶段表达明显减弱,提示其在创面上皮功能形成中的作用。角蛋白-15的Western blot分析也与krt15的转录水平一致。然而,研究发现myl-1转录水平在再生初始阶段是不可缺少的,而在分化阶段,myl-1 mRNA和蛋白水平的升高标志着其在实现再生尾鳍结构完整性方面的重要作用。这是首次观察到角蛋白-15和肌球蛋白轻链-1参与调节青鱼的再生过程。
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Temporal Expression Pattern of Peptides in the Regenerating Caudal Fin of Teleost Fish Poecilia latipinna With Special Emphasis on krt15 and myl-1
Amongst several vertebrates, caudal fin of teleost fish has been emerged as an excellent model to understand the mechanism of regeneration. By now it is well perceived that proper coordination of different regulatory signals are needed to facilitate the progression of successive stages of regeneration. Therefore, in the current study protein expression profile for caudal fin regeneration in Poecilia latipnna was brought into the focus. The pattern of protein turnover and variation in the expression level were observed across the stages of regeneration. Data from twodimensional gel electrophoresis revealed the differential expression of proteins for wound epithelium, blastema and differentiation stages when compared to resting stage. Based on the computational analysis, of the peptide expression profile and subsequent sequencing, two peptides namely Keratin type I cytoskeletal 15 and Myosin light chain-1 were studied further at transcript level. keratin-15 was found to be up regulated at wound epithelium stage while its expression waned significantly in the blastema and differentiation stages of regeneration indicating its role in the formation of a functional wound epithelium. Western blot analysis of keratin-15 also concurred with transcript levels of krt15. However, myl-1 transcript levels was observed to be dispensable for initial phase of regeneration whereas its elevated level in differentiation stage at both mRNA and protein level marks its noteworthy role in achieving the structural integrity of a regenerating caudal fin. This is the first observation of the involvement of keratin-15 and Myosin light chain-1 in the regulation of regenerative process in Poecilia latipinna.
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