Long-Term Exposure of RN46A Cells Expressing Serotonin Transporter (SERT) to a cAMP Analog Up-regulates SERT Activity and Is Accompanied by Neural Differentiation of the Cells
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概要
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To examine the functional regulation of serotonin transporter (SERT) by cAMP, we examined whether SERT uptake activity was affected by dibutyryl cAMP (dbcAMP), a cAMP analog, in SERT-transfected RN46A cells derived from embryonic rat raphe neurons. Long-term exposure (> 4 h) of dbcAMP (1 mM) to SERT-expressing RN46A cells significantly up-regulated SERT activity. In addition, a selective PKA activator, but not a selective EPAC activator, increased the serotonin uptake activity of SERT, suggesting that this regulation was mainly mediated via PKA. Time-dependent up-regulation of SERT activity by dbcAMP was accompanied by neural differentiation of RN46A cells. Further investigation of dbcAMP-induced up-regulation of SERT revealed that dbcAMP elevated SERT protein levels without affecting SERT mRNA transcription. The chase assay for residual SERT protein revealed that dbcAMP slowed its degradation rate. Immunohistochemical analysis revealed that plasma membrane–localized SERT was more abundant in dbcAMP-treated cells than in non-treated cells, suggesting that dbcAMP up-regulated SERT by decreasing its degradation and increasing its plasma membrane expression. These results raise the possibility that the elevation of intracellular cAMP up-regulated SERT function through a mechanism linked to the differentiation of RN46A cells and show the importance of SERT function during the developmental process of the serotonergic nervous system.
著者
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Hide Izumi
Department Of Molecular And Pharmacological Neuroscience Graduate School Of Biomedical Sciences Hiro
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Tanaka Anna
Department Of Molecular And Applied Bioscience Graduate School Of Biosphere Science Hiroshima Univer
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Sakai Norio
Department Of Biopharmaceutics And Drug Rational Research Center School Of Pharmacy Tokyo University
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Tanaka Shigeru
Department Of Applied Chemistry Faculty Of Science And Technology Keio University
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Seki Takahiro
Department Of Applied Physics Nagoya University
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Seki Takahiro
Department of Molecular and Pharmacological Neuroscience, Institute of Biomedical & Health Sciences, Hiroshima University, Japan
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Yammamoto Hikaru
Department of Molecular and Pharmacological Neuroscience, Institute of Biomedical & Health Sciences, Hiroshima University, Japan
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Tanaka Anna
Department of Molecular and Pharmacological Neuroscience, Institute of Biomedical & Health Sciences, Hiroshima University, Japan
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Yammamoto Hikaru
Department of Molecular and Pharmacological Neuroscience, Institute of Biomedical & Health Sciences, Hiroshima University, Japan
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