Isolation and functional characterization of an allatotropin receptor from Manduca sexta
Isolation and functional characterization of an allatotropin receptor from Manduca sexta
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DOI：
10.1016/j.ibmb.2011.06.002
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发表时间：
2011-10-01
影响因子：
3.8
通讯作者：
Broeck, Jozef Vanden
中科院分区：
文献类型：
--
作者：
Horodyski, Frank M.;Verlinden, Heleen;Broeck, Jozef Vanden
Manduca sexta allatotroin(Manse-AT)是一种多功能神经肽，其作用包括刺激保幼激素的生物合成、肌力刺激、心脏加速功能和抑制主动离子转运。MANSE-AT是一个结构相关的多肽家族的成员，该家族广泛存在于昆虫和其他无脊椎动物中。它的确切作用取决于昆虫的种类和发育阶段。在一些鳞翅目昆虫中，结构相关的AT样多肽可以从AT基因转录的选择性剪接的mRNAs中获得。我们利用家蚕AT受体(ATR)的序列，通过基于聚合酶链式反应(PCR)的方法，从家蚕中分离到了AT受体(AIR)的cDNA。Sexta ATR的序列与其他昆虫的G蛋白偶联受体以及哺乳动物的食欲素受体相似。我们证明在脊椎动物细胞系中表达的六分枝杆菌ATR被Manse-AT和每个Manse-an肽以剂量反应的方式激活，顺序为ATL-I>；AN-II>；ATL-III>；AT，并且在多个细胞系中的功能分析表明，该受体是通过升高的钙和cAMP水平偶联的。在食性幼虫中，Manse-ATR mRNA在马氏管中表达水平最高，其次是中肠、后肠、睾丸和鼻侧体部，这与其对多个靶组织的作用一致。在成人的心体-翼体复合体中，Manse-air mRNA在两性中的表达水平都相对较低。(C)2011爱思唯尔有限公司。保留所有权利。
Manduca sexta allatotropin (Manse-AT) is a multifunctional neuropeptide whose actions include the stimulation of juvenile hormone biosynthesis, myotropic stimulation, cardioacceleratory functions, and inhibition of active ion transport. Manse-AT is a member of a structurally related peptide family that is widely found in insects and also in other invertebrates. Its precise role depends on the insect species and developmental stage. In some lepidopteran insects including M. sexta, structurally-related AT-like (An) peptides can be derived from alternatively spliced mRNAs transcribed from the AT gene. We have isolated a cDNA for an AT receptor (AIR) from M. sexta by a PCR-based approach using the sequence of the ATR from Bombyx mori. The sequence of the M. sexta ATR is similar to several G protein-coupled receptors from other insect species and to the mammalian orexin receptor. We demonstrate that the M. sexta ATR expressed in vertebrate cell lines is activated in a dose-responsive manner by Manse-AT and each Manse-An peptide in the rank order ATL-I > An-II > ATL-III > AT, and functional analysis in multiple cell lines suggest that the receptor is coupled through elevated levels of Ca2+ and cAMP. In feeding larvae, Manse-ATR mRNA is present at highest levels in the Malpighian tubules, followed by the midgut, hindgut, testes, and corpora allata, consistent with its action on multiple target tissues. In the adult corpora cardiaca corpora allata complex, Manse-AIR mRNA is present at relatively low levels in both sexes. (C) 2011 Elsevier Ltd. All rights reserved.