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.

中文翻译：

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蝾螈口咽外植体在味蕾分化阶段的转录组分析

墨西哥蝾螈 Ambystoma mexicanum (Axolotl) 是了解发育和再生的优秀脊椎动物模型系统。蝾螈胚胎的研究为味蕾发育提供了重要的见解。在原肠胚形成和神经形成过程中，蝾螈胚胎的口咽内胚层中分别发生味蕾特化和决定，而味蕾神经支配和味觉细胞分化发生在发育后期。Axolotl 胚胎适合显微外科手术，组织外植体很容易在体外发育。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 管道来执行测序读数的从头组装。RNA-seq 数据的线性模型用于鉴定差异表达的基因。我们发现了 1234 个在味觉细胞分化阶段差异表达的独特基因。我们使用 RTqPCR 验证了其中四个基因并进行了 GO 功能分析。这些基因的差异表达表明它们可能在蝾螈的味觉细胞分化中起作用。