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从西方棉叶蝉转录组数据中挖掘瞬时受体电位通道:痛觉缺失同源物的表达谱和功能特征。

Mining Lygus hesperus (western tarnished plant bug) transcriptomic data for transient receptor potential channels: Expression profiling and functional characterization of a Painless homolog.

机构信息

Pest Management and Biocontrol Research Unit, US Arid Land Agricultural Research Center, USDA Agricultural Research Services, Maricopa, AZ 85138, USA.

Pest Management and Biocontrol Research Unit, US Arid Land Agricultural Research Center, USDA Agricultural Research Services, Maricopa, AZ 85138, USA.

出版信息

Comp Biochem Physiol Part D Genomics Proteomics. 2022 Dec;44:101027. doi: 10.1016/j.cbd.2022.101027. Epub 2022 Oct 7.

Abstract

The transient receptor potential (TRP) family of cation channels are evolutionarily conserved proteins with critical roles in sensory physiology. Despite extensive studies in model species, knowledge of TRP channel functional diversity and physiological impact remains limited in many non-model insect species. To assess the TRP channel repertoire in a non-model agriculture pest species (Lygus hesperus), publicly available transcriptomic datasets were mined for potential homologs. Among the transcripts identified, 30 are predicted to encompass complete open reading frames that encode proteins representing each of the seven TRP channel subfamilies. Although no homologs were identified for the Pyrexia and Brivido channels, the TRP complement in L. hesperus exceeded the 13-16 channels reported in most insects. This diversity appears to be driven by a combination of alternative splicing, which impacted members of six subfamilies, and gene expansion of the TRPP subfamily. To validate the in silico data and provide more detailed analyses of L. hesperus TRP functionality, the putative Painless homolog was selected for more in depth analysis and its functional role in thermosensation examined in vitro. RT-PCR expression profiling revealed near ubiquitous expression of the Painless transcript throughout nymphal and adult development. Electrophysiological data generated using a Xenopus oocyte recombinant expression system indicated activation parameters for L. hesperus Painless homolog that are consistent with a role in noxious heat (40°-45 °C) thermosensation.

摘要

瞬时受体电位 (TRP) 阳离子通道家族是进化上保守的蛋白质,在感觉生理学中具有关键作用。尽管在模式物种中进行了广泛的研究,但在许多非模式昆虫物种中,TRP 通道功能多样性和生理影响的知识仍然有限。为了评估非模式农业害虫物种(褐盲蝽)中的 TRP 通道谱,从公开的转录组数据集挖掘潜在的同源物。在所鉴定的转录本中,有 30 个预测包含完整的开放阅读框,编码代表七个 TRP 通道亚家族的每个亚家族的蛋白质。虽然没有鉴定出 Pyrexia 和 Brivido 通道的同源物,但褐盲蝽中的 TRP 组成超过了大多数昆虫报告的 13-16 个通道。这种多样性似乎是由剪接的替代,影响了六个亚家族的成员,以及 TRPP 亚家族的基因扩张共同驱动的。为了验证计算机数据并提供对褐盲蝽 TRP 功能的更详细分析,选择了假定的 Painless 同源物进行更深入的分析,并在体外检查其在热感觉中的功能作用。RT-PCR 表达谱分析显示,Painless 转录本在若虫和成虫发育过程中的近乎普遍表达。使用非洲爪蟾卵母细胞重组表达系统生成的电生理数据表明,褐盲蝽 Painless 同源物的激活参数与其在有害热(40°-45°C)热感觉中的作用一致。

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