Identification of an allatostatin C signaling system in mollusc Aplysia.
Identification of an allatostatin C signaling system in mollusc Aplysia.
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DOI：
10.1038/s41598-022-05071-8
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发表时间：
2022-01-24
影响因子：
4.6
通讯作者：
Jing J
中科院分区：
文献类型：
--
作者：
Jiang HM;Yang Z;Xue YY;Wang HY;Guo SQ;Xu JP;Li YD;Fu P;Ding XY;Yu K;Liu WJ;Zhang G;Wang J;Zhou HB;Susswein AJ;Jing J
神经肽作为中枢神经系统中普遍存在的细胞间信号分子，调节着原口动物和后口动物的多种行为系统。allatostins是节肢动物中抑制幼年激素生物合成的神经肽。allatostatin C （allatostatin C, AstC）首次从Manduca sexta中分离得到，具有由两个半胱氨酸残基组成的二硫桥的重要保守特征。此外，AstC似乎是哺乳动物生长抑素的同源物，它与生长抑素具有共同的功能，如调节摄食行为。AstC信号系统在节肢动物中被广泛研究，但在软体动物中的研究很少。在这项研究中，我们试图识别海洋软体动物加州海陆的AstC信号系统。我们从白头草的cDNA中克隆出了AstC前体。我们使用NeuroPred预测了一个含有15个氨基酸的二硫桥肽，即AstC。随后，我们克隆了两个推测的抑素c样受体，并通过NCBI保守结构域搜索发现它们属于G蛋白偶联受体（GPCR）家族。此外，通过肌醇单磷酸1 （IP1）积累实验，我们发现apilsia AstC可以以最低的EC50激活其中一种假定的受体，即AstC- r，而没有二硫桥接的AstC （AstC'）激活的AstC- r具有最高的EC50。此外，4种软体动物的AstC序列与大灰鲸的AstC序列不同，但具有二硫桥，可以在中间EC50激活AstC- r。综上所述，我们成功鉴定了海兔AstC前体及其受体（AstC- r），这在软体动物中是首例，为进一步研究海兔及其他软体动物的AstC信号系统提供了重要的基础。
Neuropeptides, as pervasive intercellular signaling molecules in the CNS, modulate a variety of behavioral systems in both protostomes and deuterostomes. Allatostatins are neuropeptides in arthropods that inhibit the biosynthesis of juvenile hormones. Based on amino acid sequences, they are divided into three different types in arthropods: allatostatin A, allatostatin B, allatostatin C. Allatostatin C (AstC) was first isolated from Manduca sexta, and it has an important conserved feature of a disulfide bridge formed by two cysteine residues. Moreover, AstC appears to be the ortholog of mammalian somatostatin, and it has functions in common with somatostatin, such as modulating feeding behaviors. The AstC signaling system has been widely studied in arthropods, but minimally studied in molluscs. In this study, we seek to identify the AstC signaling system in the marine mollusc Aplysia californica. We cloned the AstC precursor from the cDNA of Aplysia. We predicted a 15-amino acid peptide with a disulfide bridge, i.e., AstC, using NeuroPred. We then cloned two putative allatostatin C-like receptors and through NCBI Conserved Domain Search we found that they belonged to the G protein-coupled receptor (GPCR) family. In addition, using an inositol monophosphate 1 (IP1) accumulation assay, we showed that Aplysia AstC could activate one of the putative receptors, i.e., the AstC-R, at the lowest EC50, and AstC without the disulfide bridge (AstC') activated AstC-R with the highest EC50. Moreover, four molluscan AstCs with variations of sequences from Aplysia AstC but with the disulfide bridge activated AstC-R at intermediate EC50. In summary, our successful identification of the Aplysia AstC precursor and its receptor (AstC-R) represents the first example in molluscs, and provides an important basis for further studies of the AstC signaling system in Aplysia and other molluscs.
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