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蝾螈口咽部外植体在味蕾分化阶段的转录组分析。

Transcriptome analysis of axolotl oropharyngeal explants during taste bud differentiation stages.

机构信息

Department of Mathematics and Statistics, Connecticut College, 270 Mohegan Avenue, New London, CT, United States of America.

Department of Biology, Connecticut College, 270 Mohegan Avenue, New London, CT, United States of America.

出版信息

Mech Dev. 2020 Mar;161:103597. doi: 10.1016/j.mod.2020.103597. Epub 2020 Feb 7.

Abstract

The Mexican salamander, Ambystoma mexicanum (Axolotl), is an excellent vertebrate model system to understand development and regeneration. Studies in axolotl embryos have provided important insights into taste bud development. Taste bud specification and determination occur in the oropharyngeal endoderm of axolotl embryos during gastrulation and neurulation, respectively, whereas taste bud innervation and taste cell differentiation occur later in development. Axolotl embryos are amenable to microsurgery, and tissue explants develop readily in vitro. We performed RNA-seq analysis to investigate the differential expression of genes in oropharyngeal explants at several stages of taste cell differentiation. Since the axolotl genome has only recently been sequenced, we used a Trinity pipeline to perform de novo assembly of sequencing reads. Linear models for RNA-seq data were used to identify differentially expressed genes. We found 1234 unique genes differentially expressed during taste cell differentiation stages. We validated four of these genes using RTqPCR and performed GO functional analysis. The differential expression of these genes suggests that they may play a role in taste cell differentiation in axolotls.

摘要

墨西哥钝口螈,Ambystoma mexicanum(Axolotl),是一种优秀的脊椎动物模型系统,可用于研究发育和再生。在 Axolotl 胚胎中的研究为研究味蕾发育提供了重要的见解。味蕾的特化和决定分别发生在 Axolotl 胚胎的口咽内胚层在原肠胚和神经胚期,而味蕾的神经支配和味觉细胞分化发生在发育后期。Axolotl 胚胎适合进行显微手术,组织外植体在体外很容易发育。我们进行了 RNA-seq 分析,以研究在味觉细胞分化的几个阶段中口咽外植体中基因的差异表达。由于 Axolotl 基因组最近才被测序,我们使用 Trinity 管道对测序reads 进行从头组装。用于 RNA-seq 数据的线性模型用于鉴定差异表达的基因。我们发现 1234 个独特的基因在味觉细胞分化阶段差异表达。我们使用 RTqPCR 验证了其中的四个基因,并进行了 GO 功能分析。这些基因的差异表达表明它们可能在 Axolotl 中味觉细胞分化中发挥作用。

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