Alternatively spliced orcokinin isoforms and their functions in Tribolium castaneum.
Alternatively spliced orcokinin isoforms and their functions in Tribolium castaneum.
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DOI：
10.1016/j.ibmb.2015.07.009
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发表时间：
2015-10
影响因子：
3.8
通讯作者：
Park Y
中科院分区：
文献类型：
--
作者：
Jiang H;Kim HG;Park Y
Orcokinin和orcomyotropin最初被描述为甲壳类动物中的神经肽，但现在已经在许多昆虫物种中发现，它们分别被称为orcokininin-A（OK-A）和orcokinin-B（OK-B）。这两组成熟肽是昆虫中单拷贝基因orcokinin的选择性剪接转录产物。本文研究了赤拟谷盗OK-A和OK-B的表达模式及其功能。原位杂交和免疫组织化学使用同种型特异性探针和抗体的每一个OK-A和OK-B表明，这两种肽共表达在5至7对脑细胞和中肠肠内分泌细胞，这与其他昆虫的表达模式，其中两种肽在不同的细胞中表达。我们开发了一种新的行为检测方法来评估T.栗色。单独或联合使用ok-a和ok-b的RNAi在成虫试验中导致响应于机械刺激的更高频率和更长持续时间的假死。在幼虫行为测定中，我们观察到在用单独或组合的ok-a和ok-b的RNAi处理的昆虫中，从水浸诱导的敲除恢复时间更长。我们的结论是，OK-A和OK-B都有“觉醒”活动，并可能参与昼夜节律的控制。
Orcokinin and orcomyotropin were originally described as neuropeptides in crustaceans but have now been uncovered in many species of insects in which they are called orcokinin-A (OK-A) and orcokinin-B (OK-B), respectively. The two groups of mature peptides are products of alternatively spliced transcripts of the single copy gene orcokinin in insects. We investigated the expression patterns and the functions of OK-A and OK-B in the red flour beetle Tribolium castaneum. In situ hybridization and immunohistochemistry using isoform-specific probes and antibodies for each OK-A and OK-B suggests that both peptides are co-expressed in 5 to 7 pairs of brain cells and in the midgut enteroendocrine cells, which contrasts to expression patterns in other insects in which the two peptides are expressed in different cells. We developed a novel behavioral assay to assess the phenotypes of orcokinin RNA interference (RNAi) in T. castaneum. RNAi of ok-a and ok-b alone or in combination resulted in higher frequencies and longer durations of death feigning in response to mechanical stimulation in the adult assay. In the larval behavioral assays, we observed longer recovery times from knockout induced by water submergence in the insects treated with RNAi for ok-a and ok-b alone or in combination. We conclude that both OK-A and OK-B have “awakening” activities and are potentially involved in the control of circadian rhythms.