Expression Changes of MHC and Other Immune Genes in Frog Skin during Ontogeny
Expression Changes of MHC and Other Immune Genes in Frog Skin during Ontogeny
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DOI：
10.3390/ani10010091
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发表时间：
2020-01
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影响因子：
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通讯作者：
Q. Lau;Takeshi Igawa;Shohei Komaki;Y. Satta
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作者：
Q. Lau;Takeshi Igawa;Shohei Komaki;Y. Satta
蝌蚪在蜕变为成年蛙之前，在生理和免疫方面经历了许多变化。主要组织相容性复合体（MHC）分子是脊椎动物适应性免疫的重要组成部分，本研究测量了山褐蛙（Rana ornativentris）六个蝌蚪期皮肤中两种MHC基因（MHC I类和MHC II类）的表达。首先，使用qPCR方法，我们发现MHC I类和II类表达在24/25期（“早期”）和28期（“中期”）蝌蚪皮肤之间显著增加。然后，我们对R. ornativentris和模式物种热带爪蟾（Xenopus tropicalis）的“早期”、“中期”和“晚期”蝌蚪皮肤mRNA进行了下一代测序，并证实MHC表达从“中期”开始增加。我们还对转录组数据进行了进一步分析，发现几个与免疫相关的基因本体术语从蝌蚪中期开始上调。我们的研究结果可能支持MHC I类和II类在蝌蚪发育过程中都有功能作用。摘要：无尾两栖动物在变态后经历了重大的生理和免疫变化。主要组织相容性复合体（MHC）的基因编码对脊椎动物适应性免疫很重要的受体。我们使用qPCR方法测量了6个不同本体分期（n = 10 /分期）的皮肤MHC表达；28期（中期幼虫）与24/25期（早期幼虫）蝌蚪相比，标准化MHC I类和II类mRNA水平的表达显著升高。随后对三种蝌蚪（早期、中期和晚期幼虫）的转录组学分析聚焦于皮肤中免疫相关基因的mRNA表达。与早期幼虫相比，这两个物种中大多数MHC I类和II类转录本的标准化表达在幼虫中期和后期显著更高。此外，对差异表达转录本的基因本体论（GO）分析显示，从幼虫中期开始，一些与免疫相关的GO术语显著上调。我们的研究提供了MHC I类和II类在花椒和热带蓟的发育过程中表达的证据。
Simple Summary Tadpoles undergo many changes in physiology and immunology until metamorphosis into adult frogs. Major histocompatibility complex (MHC) molecules are an important part of vertebrate adaptive immunity, and our study measured the expression of two MHC genes (MHC class I and II) in skin during six tadpole stages of the Montane Brown frog (Rana ornativentris). First, using a qPCR method, we found that both MHC class I and II expression significantly increased between stage 24/25 (‘early’) and stage 28 (‘mid’) tadpole skin. Then, we conducted next-generation sequencing for ‘early’, ‘mid’ and ‘late’ stage tadpole skin mRNA of both R. ornativentris and a model species, Xenopus tropicalis, and confirmed that MHC expression increased from the ‘mid’ stage. We also performed further analyses of transcriptome data and found that several immune-related gene ontology terms were upregulated from the ‘mid’ tadpole stage. Our findings probably support that both MHC class I and II have a functional role during tadpole development. Abstract Anuran amphibians undergo major physiological and immunological changes following metamorphosis. Genes of the major histocompatibility complex (MHC) code for receptors important for vertebrate adaptive immunity. We used qPCR to measure skin MHC expression in six different ontological stages of Rana ornativentris (n = 10 per stage); normalized MHC class I and II expression at the mRNA level was significantly higher in stage 28 (mid-larval) compared to stages 24/25 (early-larval) tadpoles. Subsequent transcriptomic analyses of three tadpole (early-, mid-, and late-larval) stages of R. ornativentris and model species Xenopus tropicalis focused on mRNA expression of immune-related genes in the skin. Normalized expression of most MHC class I and II transcripts in both species were significantly higher in mid- and late-larval stages compared to early-larval stage. In addition, gene ontology (GO) analyses of differentially expressed transcripts revealed several immune-related GO terms that were significantly upregulated from the mid-larval stage. Our study provides evidence that both MHC class I and II is expressed during development in both R. ornativentris and X. tropicalis.