神经肽 RYamides 激活家蚕神经肽 G 蛋白偶联受体 A19，偶联 Gq 蛋白依赖性信号通路,Journal of Cellular Biochemistry

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神经肽 RYamides 激活家蚕神经肽 G 蛋白偶联受体 A19，偶联 Gq 蛋白依赖性信号通路
Journal of Cellular Biochemistry ( IF 3.0 ) Pub Date : 2021-01-05 , DOI: 10.1002/jcb.29874
Zhiqiang Guo ₁ , Xiaobai He ₂ , Chaohui Jiang ₁ , Ying Shi ₁ , Naiming Zhou ₁

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RYamides 构成了在昆虫中新发现的一个新的神经肽家族，在调节各种生理过程中发挥着重要作用。然而，RYamide 信号系统的信号特征和生理作用仍然很大程度上未知。在本研究中，我们从家蚕幼虫中克隆了 RYamide 受体 BNGR-A19 的全长互补 DNA 。在哺乳动物 HEK293T 和昆虫 Sf9 细胞中表达后，功能分析表明 BNGR-A19 被合成的 RYamide 肽激活，触发 cAMP 反应元件控制的荧光素酶活性和G _{q 中}Ca ²⁺动员的显着增加抑制剂敏感的方式。在被 RYamide 肽激活后，BNGR-A19 通过 G _q -PLC-PKC 途径引发 ERK1/2 磷酸化，并且还经历了从细胞表面到细胞质的快速内化。进一步的交叉活性分析表明，BNGR-A19 在被高浓度 (1 μM) 的家蚕sulfakin-1、神经肽 F-1 和短神经肽 F-1刺激时表现出非常弱的反应，反之亦然，家蚕RYamides 也表现出轻微的激活Bombyx 的效力NPF 受体 (BNGR-A4) 和 sNPF 受体 (BNGR-A11)。此外，定量逆转录聚合酶链反应结果表明，在后肠和睾丸中检测到 BNGR-A19 的高水平表达，表明 RYamide 信号可能参与了摄食、水分稳态和睾丸发育的调节. 该研究首次提供了有关昆虫 RYamide 信号系统的深入信息，有助于进一步阐明其在昆虫生理学中的内分泌作用。

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Activation of Bombyx mori neuropeptide G protein–coupled receptor A19 by neuropeptide RYamides couples to Gq protein‐dependent signaling pathways

RYamides constitute a novel family of neuropeptides newly identified in insects, and play important roles in regulating a variety of physiological processes. However, the signaling characteristics and physiological actions of RYamide signaling system remain largely unknown. In the present study, we cloned the full‐length complementary DNA of the RYamide receptor BNGR‐A19 from Bombyx mori larvae. After expression in mammalian HEK293T and insect Sf9 cells, functional assays revealed that BNGR‐A19 was activated by synthetic RYamide peptides, triggering a significant increase in cAMP‐response element controlled luciferase activity and Ca²⁺ mobilization in a G_q inhibitor‐sensitive manner. Upon activation by RYamide peptides, BNGR‐A19 elicited ERK1/2 phosphorylation via a G_q‐PLC‐PKC pathway, and also underwent a rapid internalization from the cell surface to the cytoplasm. Further cross‐activity analysis indicated that BNGR‐A19 exhibited very weak response upon stimulation by high concentration (1 μM) of Bombyx sulfakinin‐1, neuropeptide F‐1, and short neuropeptide F‐1, and vice versa, Bombyx RYamides also showed slight potency for activating Bombyx NPF receptor (BNGR‐A4) and sNPF receptor (BNGR‐A11). In addition, the quantitative reverse‐transcription polymerase chain reaction results showed that the high‐level expression of BNGR‐A19 was detected in the hindgut and testis, suggesting that the RYamide signaling is likely involved in the regulation of feeding, water homeostasis and testis development. This study provides the first in‐depth information on the insect RYamide signaling system, facilitating the further clarification of its endocrinological roles in insect physiology.

更新日期：2021-02-16

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