Nakamura Kenta, Islam Md Rafiqul, Takayanagi Miyako, Yasumuro Hirofumi, Inami Wataru, Kunahong Ailidana, Casco-Robles Roman M, Toyama Fubito, Chiba Chikafumi
Faculty of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, Japan.
Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, Japan.
PLoS One. 2014 Oct 7;9(10):e109831. doi: 10.1371/journal.pone.0109831. eCollection 2014.
Retinal regeneration in the adult newt is a useful system to uncover essential mechanisms underlying the regeneration of body parts of this animal as well as to find clues to treat retinal disorders such as proliferative vitreoretinopathy. Here, to facilitate the study of early processes of retinal regeneration, we provide a de novo assembly transcriptome and inferred proteome of the Japanese fire bellied newt (Cynops pyrrhogaster), which was obtained from eyeball samples of day 0-14 after surgical removal of the lens and neural retina. This transcriptome (237,120 in silico transcripts) contains most information of cDNAs/ESTs which has been reported in newts (C. pyrrhogaster, Pleurodeles waltl and Notophthalmus viridescence) thus far. On the other hand, de novo assembly transcriptomes reported lately for N. viridescence only covered 16-31% of this transcriptome, suggesting that most constituents of this transcriptome are specific to the regenerating eye tissues of C. pyrrhogaster. A total of 87,102 in silico transcripts of this transcriptome were functionally annotated. Coding sequence prediction in combination with functional annotation revealed that 76,968 in silico transcripts encode protein/peptides recorded in public databases so far, whereas 17,316 might be unique. qPCR and Sanger sequencing demonstrated that this transcriptome contains much information pertaining to genes that are regulated in association with cell reprogramming, cell-cycle re-entry/proliferation, and tissue patterning in an early phase of retinal regeneration. This data also provides important insight for further investigations addressing cellular mechanisms and molecular networks underlying retinal regeneration as well as differences between retinal regeneration and disorders. This transcriptome can be applied to ensuing comprehensive gene screening steps, providing candidate genes, regardless of whether annotated or unique, to uncover essential mechanisms underlying early processes of retinal regeneration.
成年蝾螈的视网膜再生是一个有用的系统,可用于揭示该动物身体部位再生的基本机制,以及寻找治疗视网膜疾病(如增殖性玻璃体视网膜病变)的线索。在此,为了便于研究视网膜再生的早期过程,我们提供了日本红腹蝾螈(Cynops pyrrhogaster)的从头组装转录组和推断蛋白质组,这些数据来自于手术摘除晶状体和神经视网膜后0-14天的眼球样本。这个转录组(237,120个电子转录本)包含了迄今为止在蝾螈(C. pyrrhogaster、Pleurodeles waltl和Notophthalmus viridescence)中报道的大部分cDNA/EST信息。另一方面,最近报道的N. viridescence的从头组装转录组仅覆盖了这个转录组的16-31%,这表明这个转录组的大多数成分是C. pyrrhogaster再生眼组织特有的。这个转录组共有87,102个电子转录本被进行了功能注释。结合功能注释的编码序列预测表明,76,968个电子转录本编码了目前在公共数据库中记录的蛋白质/肽,而17,316个可能是独特的。qPCR和桑格测序表明,这个转录组包含了许多与视网膜再生早期阶段中与细胞重编程、细胞周期重新进入/增殖以及组织模式相关的基因调控信息。这些数据也为进一步研究视网膜再生的细胞机制和分子网络以及视网膜再生与疾病之间的差异提供了重要的见解。这个转录组可应用于后续的全面基因筛选步骤,提供候选基因,无论其是否被注释或独特,以揭示视网膜再生早期过程的基本机制。
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