Does Differential Receptor Distribution Underlie Variable Responses to a Neuropeptide in the Lobster Cardiac System?
Does Differential Receptor Distribution Underlie Variable Responses to a Neuropeptide in the Lobster Cardiac System?
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DOI：
10.3390/ijms22168703
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发表时间：
2021-08-13
影响因子：
5.6
通讯作者：
Dickinson PS
中科院分区：
文献类型：
--
作者：
Muscato AJ;Walsh P;Pong S;Pupo A;Gross RJ;Christie AE;Hull JJ;Dickinson PS
中枢模式生成器产生独立于感觉输入的有节奏的行为；然而，它们的输出可以被神经肽调节，从而允许功能灵活性。我们研究了C型生长抑素(AST-C)对心脏神经节(CG)的影响，心脏神经节是控制美洲龙虾心脏的中央模式发生器，以确定个体对AST-C反应显著差异的生物学机制。我们提出，多个受体的存在，从而不同的受体分布，至少是这种观察到的差异性的部分原因。利用转录组挖掘和基于PCR的克隆，我们在CG中鉴定了四个AST-C受体(ASTCRs)；然后我们研究了它们的细胞定位、结合潜力和功能激活。在我们的昆虫细胞表达系统中，只有两个受体ASTCR1和ASTCR2是全功能的GPCRs，它们定位于细胞表面，并被AST-C多肽激活。然而，这四个基因都是从CG的cDNA中扩增出来的。在ASTCR表达得到确认后，我们使用生理学和生物信息学技术将受体表达与个体间心脏对AST-C的反应联系起来。ASTCR1在CG中的表达与通过龙虾心脏对AST-C灌流的收缩幅度增加呈负相关，这表明CG中ASTCRs的差异表达可能是特定龙虾对AST-C表现出的特异性生理反应的部分原因。
Central pattern generators produce rhythmic behaviors independently of sensory input; however, their outputs can be modulated by neuropeptides, thereby allowing for functional flexibility. We investigated the effects of C-type allatostatins (AST-C) on the cardiac ganglion (CG), which is the central pattern generator that controls the heart of the American lobster, Homarus americanus, to identify the biological mechanism underlying the significant variability in individual responses to AST-C. We proposed that the presence of multiple receptors, and thus differential receptor distribution, was at least partly responsible for this observed variability. Using transcriptome mining and PCR-based cloning, we identified four AST-C receptors (ASTCRs) in the CG; we then characterized their cellular localization, binding potential, and functional activation. Only two of the four receptors, ASTCR1 and ASTCR2, were fully functional GPCRs that targeted to the cell surface and were activated by AST-C peptides in our insect cell expression system. All four, however, were amplified from CG cDNAs. Following the confirmation of ASTCR expression, we used physiological and bioinformatic techniques to correlate receptor expression with cardiac responses to AST-C across individuals. Expression of ASTCR1 in the CG showed a negative correlation with increasing contraction amplitude in response to AST-C perfusion through the lobster heart, suggesting that the differential expression of ASTCRs within the CG is partly responsible for the specific physiological response to AST-C exhibited by a given individual lobster.
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