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原位杂交分析揭示海兔磷酸二酯酶4在袋状细胞中的特异性表达

Specific Expression of Aplysia Phosphodiesterase 4 in Bag Cells Revealed by in situ Hybridization Analysis.

作者信息

Jang Deok-Jin, Kim Hyoung F, Sim Jae-Hoon, Lim Chae-Seok, Kaang Bong-Kiun

机构信息

Department of Ecological Science, College of Ecology and Environment, Kyungpook National University, Sangju 37224, Korea.

Department of Biological Sciences, College of Natural Sciences, Seoul National University, Seoul 08826, Korea.

出版信息

Exp Neurobiol. 2015 Sep;24(3):246-51. doi: 10.5607/en.2015.24.3.246. Epub 2015 Aug 13.

Abstract

Phosphodiesterases (PDEs) play a key role in the regulation of cyclic adenosine monophosphate (cAMP), which in turn mediates various cellular functions including learning and memory. We previously cloned and characterized three PDE4 isoforms (ApPDE4) from Aplysia kurodai. Using reverse transcription polymerase chain reaction (RT-PCR), we found that ApPDE4 isoforms are primarily expressed in the central nervous system. However, the detailed distribution of ApPDE4 mRNA in Aplysia individual ganglions was not evident. In this study, to determine the distribution of ApPDE4 mRNAs in Aplysia ganglions, we performed in situ hybridization (ISH) using a probe targeting ApPDE4, including the PDE catalytic domain. Interestingly, we found the strongest ISH-positive signals in the symmetrical bag cell clusters of the abdominal ganglion. The R2, R14, L7, L2 and L11 neurons in the abdominal ganglion, LP1 neuron in pleural ganglion, and metacerebral (MCC) neurons were ISH-positive. Mechanosensory neurons of the sensory cluster were also stained on the ventral aspect of the right and left pleural ganglia. Taken together, we found the detailed distribution of ApPDE4 mRNA in Aplysia ganglion and support their roles in serotonin (5-HT)-induced synaptic facilitation of Aplysia mechanosensory neurons.

摘要

磷酸二酯酶(PDEs)在环磷酸腺苷(cAMP)的调节中起关键作用,而cAMP反过来介导包括学习和记忆在内的各种细胞功能。我们之前从黑海天牛中克隆并鉴定了三种PDE4亚型(ApPDE4)。使用逆转录聚合酶链反应(RT-PCR),我们发现ApPDE4亚型主要在中枢神经系统中表达。然而,ApPDE4 mRNA在海兔单个神经节中的详细分布并不明显。在本研究中,为了确定ApPDE4 mRNA在海兔神经节中的分布,我们使用针对ApPDE4(包括PDE催化结构域)的探针进行了原位杂交(ISH)。有趣的是,我们在腹神经节的对称袋状细胞簇中发现了最强的ISH阳性信号。腹神经节中的R2、R14、L7、L2和L11神经元、胸膜神经节中的LP1神经元以及大脑后叶(MCC)神经元呈ISH阳性。感觉簇的机械感觉神经元在左右胸膜神经节的腹侧也被染色。综上所述,我们发现了ApPDE4 mRNA在海兔神经节中的详细分布,并支持它们在5-羟色胺(5-HT)诱导的海兔机械感觉神经元突触易化中的作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6450/4580752/bf85e048cc65/en-24-246-g001.jpg

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