Serotonin-2C receptor involved serotonin-induced Ca 2+ mobilisations in neuronal progenitors and neurons in rat suprachiasmatic nucleus

Kouhei Takeuchi; Shahid Mohammad; Tomoya Ozaki; Eri Morioka; Kaori Kawaguchi; Juhyon Kim; Byeongha Jeong; Jin Hee Hong; Kyoung J. Lee; Masayuki Ikeda

doi:10.1038/srep04106

Serotonin-2C receptor involved serotonin-induced Ca 2+ mobilisations in neuronal progenitors and neurons in rat suprachiasmatic nucleus

Kouhei Takeuchi, Shahid Mohammad, Tomoya Ozaki, Eri Morioka, Kaori Kawaguchi, Juhyon Kim, Byeongha Jeong, Jin Hee Hong, Kyoung J. Lee, Masayuki Ikeda

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Abstract

The hypothalamic suprachiasmatic nucleus (SCN), the central circadian pacemaker in mammals, undergoes serotonergic regulation, but the underlying mechanisms remain obscure. Here, we generated a subclone of an SCN progenitor cell line expressing Ca 2+ sensors (SCN2.2YC) and compared its 5-HT receptor signalling with that of rat SCN neurons in brain slices. SCN2.2YC cells expressed 5-HT1A/2A/2B/2C, but not 5A/7, while all six subtypes were expressed in SCN tissues. High K + or 5-HT increased cytosolic Ca 2+ in SCN2.2YC cells. The 5-HT responses were inhibited by ritanserin and SB-221284, but resistant to WAY-100635 and RS-127445, suggesting predominant involvement of 5-HT2C for Ca 2+ mobilisations. Consistently, Ca 2+ imaging and voltage-clamp electrophysiology using rat SCN slices demonstrated post-synaptic 5-HT2C expression. Because 5-HT2C expression was postnatally increased in the SCN and 5-HT-induced Ca 2+ mobilisations were amplified in differentiated SCN2.2YC cells and developed SCN neurons, we suggest that this signalling development occurs in accordance with central clock maturations.

Original language	English
Article number	4106
Journal	Scientific reports
Volume	4
DOIs	https://doi.org/10.1038/srep04106
Publication status	Published - 2014 Feb 17

Bibliographical note

Funding Information:
We appreciate Dr. David J. Earnest (Texas A & M University) for the kind donation of SCN2.2 cells. We are also grateful to Kazuma Wada, Kurumi Yamoto and Madoka Yoshioka for their elegant technical assistance. This work was supported in part by an NRF grant (MEST; 2012R1A2A1A01008021) from the Korean Government, to K.J.L., J.H.H. and B.J. and by a Grant-in-Aid for Scientific Research (22300108) from the Ministry of Education, Culture, Sports, Science and Technology, Japan, to M.I.

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@article{a2515477bcac4d12a1c00406fd58396a,

title = "Serotonin-2C receptor involved serotonin-induced Ca 2+ mobilisations in neuronal progenitors and neurons in rat suprachiasmatic nucleus",

abstract = "The hypothalamic suprachiasmatic nucleus (SCN), the central circadian pacemaker in mammals, undergoes serotonergic regulation, but the underlying mechanisms remain obscure. Here, we generated a subclone of an SCN progenitor cell line expressing Ca 2+ sensors (SCN2.2YC) and compared its 5-HT receptor signalling with that of rat SCN neurons in brain slices. SCN2.2YC cells expressed 5-HT1A/2A/2B/2C, but not 5A/7, while all six subtypes were expressed in SCN tissues. High K + or 5-HT increased cytosolic Ca 2+ in SCN2.2YC cells. The 5-HT responses were inhibited by ritanserin and SB-221284, but resistant to WAY-100635 and RS-127445, suggesting predominant involvement of 5-HT2C for Ca 2+ mobilisations. Consistently, Ca 2+ imaging and voltage-clamp electrophysiology using rat SCN slices demonstrated post-synaptic 5-HT2C expression. Because 5-HT2C expression was postnatally increased in the SCN and 5-HT-induced Ca 2+ mobilisations were amplified in differentiated SCN2.2YC cells and developed SCN neurons, we suggest that this signalling development occurs in accordance with central clock maturations.",

author = "Kouhei Takeuchi and Shahid Mohammad and Tomoya Ozaki and Eri Morioka and Kaori Kawaguchi and Juhyon Kim and Byeongha Jeong and Hong, {Jin Hee} and Lee, {Kyoung J.} and Masayuki Ikeda",

note = "Funding Information: We appreciate Dr. David J. Earnest (Texas A & M University) for the kind donation of SCN2.2 cells. We are also grateful to Kazuma Wada, Kurumi Yamoto and Madoka Yoshioka for their elegant technical assistance. This work was supported in part by an NRF grant (MEST; 2012R1A2A1A01008021) from the Korean Government, to K.J.L., J.H.H. and B.J. and by a Grant-in-Aid for Scientific Research (22300108) from the Ministry of Education, Culture, Sports, Science and Technology, Japan, to M.I.",

year = "2014",

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day = "17",

doi = "10.1038/srep04106",

language = "English",

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T1 - Serotonin-2C receptor involved serotonin-induced Ca 2+ mobilisations in neuronal progenitors and neurons in rat suprachiasmatic nucleus

AU - Takeuchi, Kouhei

AU - Mohammad, Shahid

AU - Ozaki, Tomoya

AU - Morioka, Eri

AU - Kawaguchi, Kaori

AU - Kim, Juhyon

AU - Jeong, Byeongha

AU - Hong, Jin Hee

AU - Lee, Kyoung J.

AU - Ikeda, Masayuki

N1 - Funding Information: We appreciate Dr. David J. Earnest (Texas A & M University) for the kind donation of SCN2.2 cells. We are also grateful to Kazuma Wada, Kurumi Yamoto and Madoka Yoshioka for their elegant technical assistance. This work was supported in part by an NRF grant (MEST; 2012R1A2A1A01008021) from the Korean Government, to K.J.L., J.H.H. and B.J. and by a Grant-in-Aid for Scientific Research (22300108) from the Ministry of Education, Culture, Sports, Science and Technology, Japan, to M.I.

PY - 2014/2/17

Y1 - 2014/2/17

N2 - The hypothalamic suprachiasmatic nucleus (SCN), the central circadian pacemaker in mammals, undergoes serotonergic regulation, but the underlying mechanisms remain obscure. Here, we generated a subclone of an SCN progenitor cell line expressing Ca 2+ sensors (SCN2.2YC) and compared its 5-HT receptor signalling with that of rat SCN neurons in brain slices. SCN2.2YC cells expressed 5-HT1A/2A/2B/2C, but not 5A/7, while all six subtypes were expressed in SCN tissues. High K + or 5-HT increased cytosolic Ca 2+ in SCN2.2YC cells. The 5-HT responses were inhibited by ritanserin and SB-221284, but resistant to WAY-100635 and RS-127445, suggesting predominant involvement of 5-HT2C for Ca 2+ mobilisations. Consistently, Ca 2+ imaging and voltage-clamp electrophysiology using rat SCN slices demonstrated post-synaptic 5-HT2C expression. Because 5-HT2C expression was postnatally increased in the SCN and 5-HT-induced Ca 2+ mobilisations were amplified in differentiated SCN2.2YC cells and developed SCN neurons, we suggest that this signalling development occurs in accordance with central clock maturations.

AB - The hypothalamic suprachiasmatic nucleus (SCN), the central circadian pacemaker in mammals, undergoes serotonergic regulation, but the underlying mechanisms remain obscure. Here, we generated a subclone of an SCN progenitor cell line expressing Ca 2+ sensors (SCN2.2YC) and compared its 5-HT receptor signalling with that of rat SCN neurons in brain slices. SCN2.2YC cells expressed 5-HT1A/2A/2B/2C, but not 5A/7, while all six subtypes were expressed in SCN tissues. High K + or 5-HT increased cytosolic Ca 2+ in SCN2.2YC cells. The 5-HT responses were inhibited by ritanserin and SB-221284, but resistant to WAY-100635 and RS-127445, suggesting predominant involvement of 5-HT2C for Ca 2+ mobilisations. Consistently, Ca 2+ imaging and voltage-clamp electrophysiology using rat SCN slices demonstrated post-synaptic 5-HT2C expression. Because 5-HT2C expression was postnatally increased in the SCN and 5-HT-induced Ca 2+ mobilisations were amplified in differentiated SCN2.2YC cells and developed SCN neurons, we suggest that this signalling development occurs in accordance with central clock maturations.

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JO - Scientific reports

JF - Scientific reports

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ER -

Serotonin-2C receptor involved serotonin-induced Ca 2+ mobilisations in neuronal progenitors and neurons in rat suprachiasmatic nucleus

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