Modulation of Ca²⁺ oscillation following ischemia and nicotinic acetylcholine receptors in primary cortical neurons by high-throughput analysis

Sasaki, Tsutomu; Hisada, Sunao; Kanki, Hideaki; Nunomura, Kazuto; Lin, Bangzhong; Nishiyama, Kumiko; Kawano, Tomohito; Matsumura, Shigenobu; Mochizuki, Hideki

doi:https://doi.org/10.1038/s41598-024-77882-w

Modulation of Ca²⁺ oscillation following ischemia and nicotinic acetylcholine receptors in primary cortical neurons by high-throughput analysis

Sasaki, Tsutomu; Hisada, Sunao; Kanki, Hideaki et al.

Scientific Reports, 2024, 14, 27667

アクセス数:1,081件（2025-08-16 10:09 集計）

固定URL: https://hdl.handle.net/11094/99697

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タイトル	Modulation of Ca²⁺ oscillation following ischemia and nicotinic acetylcholine receptors in primary cortical neurons by high-throughput analysis
著者	NRID 1000020534879 1000020534879 ORCID 0000-0001-8726-978X 0000-0001-8726-978X researchmap permalink tsutomusasaki1111 tsutomusasaki1111 Scopus Author ID 7407394272 7407394272 著者名 Sasaki, Tsutomu
	著者名 Hisada, Sunao
	NRID 1000010774218 1000010774218 researchmap permalink kan1 kan1 著者名 Kanki, Hideaki
	ORCID 0000-0002-6444-3244 0000-0002-6444-3244 researchmap permalink nunomurakazuto nunomurakazuto 著者名 Nunomura, Kazuto
	ORCID 0000-0001-5877-949X 0000-0001-5877-949X researchmap permalink linbangzhong linbangzhong Scopus Author ID 57200092216 57200092216 著者名 Lin, Bangzhong
	著者名 Nishiyama, Kumiko
	著者名 Kawano, Tomohito
	NRID 1000070467413 1000070467413 著者名 Matsumura, Shigenobu
	NRID 1000090230044 1000090230044 ORCID 0000-0002-0874-7542 0000-0002-0874-7542 researchmap permalink read0207787 read0207787 Scopus Author ID 7202006099 7202006099 著者名 Mochizuki, Hideki
キーワード等	Ca²⁺ oscillations
	High-throughput screening
	Ischemia
	nAChR
	PAM
内容	Sasaki T., Hisada S., Kanki H., et al. Modulation of Ca²⁺ oscillation following ischemia and nicotinic acetylcholine receptors in primary cortical neurons by high-throughput analysis. Scientific Reports 14, 27667 (2024); https://doi.org/10.1038/s41598-024-77882-w.
抄録	Calcium oscillations in primary neuronal cultures and iPSCs have been employed to investigate arrhythmogenicity and epileptogenicity in drug development. Previous studies have demonstrated that Ca2+ influx via NMDA and nicotinic acetylcholine receptors (nAChRs) modulates Ca2+ oscillations. Nevertheless, there has been no comprehensive investigation into the impact of ischemia or nAChR-positive allosteric modulators (PAM) drugs on Ca2+ oscillations at a level that would facilitate high-throughput screening. We investigated the effects of ischemia and nAChR subtypes or nAChR PAM agonists on Ca2+ oscillations in high-density 2D and 3D-sphere primary neuronal cultures using 384-well plates with FDSS-7000. Ischemia for 1 and 2 h resulted in an increase in the frequency of Ca2+ oscillations and a decrease in their amplitude in a time-dependent manner. The NMDA and AMPA receptor inhibition significantly suppressed Ca2+ oscillation. Inhibition of NR2A or NR2B had the opposite effect on Ca oscillations. The potentiation of ischemia-induced Ca2+ oscillations was significantly inhibited by the NMDA receptor antagonist, MK-801, and the frequency of these oscillations was suppressed by the NR2B inhibitor, Ro-256981. In the 3D-neurosphere, the application of an α7nAChR agonist increased the frequency of Ca2+ oscillations, whereas the activation of α4β2 had no effect. The combination of nicotine and PNU-120596 (type II PAM) affected the frequency and amplitude of Ca2+ oscillations in a manner distinct from that of type I PAM. These systems may be useful not only for detecting epileptogenicity but also in the search for neuroprotective agents against cerebral ischemia.
出版者	Nature Research
収録物名	Scientific Reports
巻(号)	14
ページ	27667
刊行年月	2024-11-12
言語	英語
ハンドルURL	https://hdl.handle.net/11094/99697
eISSN	20452322
関連情報 (isIdenticalTo)	DOI (出版社版) https://doi.org/10.1038/s41598-024-77882-w
アクセス権	open access
権利情報	This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
出版タイプ	Version of Record
カテゴリ	学術雑誌論文 Journal Article

資源タイプ	学術雑誌論文
ローカル資源タイプ	学術雑誌論文
DCTERMS.bibliographicCitation	Scientific Reports. 2024, 14, p. 27667
DC.title	Modulation of Ca²⁺ oscillation following ischemia and nicotinic acetylcholine receptors in primary cortical neurons by high-throughput analysis
DC.creator	Sasaki, Tsutomu
	Hisada, Sunao
	Kanki, Hideaki
	Nunomura, Kazuto
	Lin, Bangzhong
	Nishiyama, Kumiko
	Kawano, Tomohito
	Matsumura, Shigenobu
	Mochizuki, Hideki
DC.publisher	Nature Research
DC.language' scheme='DCTERMS.RFC1766	英語
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2024-11-12
DC.identifier	https://doi.org/10.1038/s41598-024-77882-w
DC.subject	Ca²⁺ oscillations
	High-throughput screening
	Ischemia
	nAChR
	PAM
DC.description	Sasaki T., Hisada S., Kanki H., et al. Modulation of Ca²⁺ oscillation following ischemia and nicotinic acetylcholine receptors in primary cortical neurons by high-throughput analysis. Scientific Reports 14, 27667 (2024); https://doi.org/10.1038/s41598-024-77882-w.
DCTERMS.abstract	Calcium oscillations in primary neuronal cultures and iPSCs have been employed to investigate arrhythmogenicity and epileptogenicity in drug development. Previous studies have demonstrated that Ca2+ influx via NMDA and nicotinic acetylcholine receptors (nAChRs) modulates Ca2+ oscillations. Nevertheless, there has been no comprehensive investigation into the impact of ischemia or nAChR-positive allosteric modulators (PAM) drugs on Ca2+ oscillations at a level that would facilitate high-throughput screening. We investigated the effects of ischemia and nAChR subtypes or nAChR PAM agonists on Ca2+ oscillations in high-density 2D and 3D-sphere primary neuronal cultures using 384-well plates with FDSS-7000. Ischemia for 1 and 2 h resulted in an increase in the frequency of Ca2+ oscillations and a decrease in their amplitude in a time-dependent manner. The NMDA and AMPA receptor inhibition significantly suppressed Ca2+ oscillation. Inhibition of NR2A or NR2B had the opposite effect on Ca oscillations. The potentiation of ischemia-induced Ca2+ oscillations was significantly inhibited by the NMDA receptor antagonist, MK-801, and the frequency of these oscillations was suppressed by the NR2B inhibitor, Ro-256981. In the 3D-neurosphere, the application of an α7nAChR agonist increased the frequency of Ca2+ oscillations, whereas the activation of α4β2 had no effect. The combination of nicotine and PNU-120596 (type II PAM) affected the frequency and amplitude of Ca2+ oscillations in a manner distinct from that of type I PAM. These systems may be useful not only for detecting epileptogenicity but also in the search for neuroprotective agents against cerebral ischemia.
DC.rights	This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
DC.format	99697
citation_title	Modulation of Ca²⁺ oscillation following ischemia and nicotinic acetylcholine receptors in primary cortical neurons by high-throughput analysis
citation_author	Sasaki, Tsutomu
	Hisada, Sunao
	Kanki, Hideaki
	Nunomura, Kazuto
	Lin, Bangzhong
	Nishiyama, Kumiko
	Kawano, Tomohito
	Matsumura, Shigenobu
	Mochizuki, Hideki
citation_publisher	Nature Research
citation_language	英語
citation_date	2024-11-12
citation_journal_title	Scientific Reports
citation_volume	14
citation_firstpage	27667
citation_doi	https://doi.org/10.1038/s41598-024-77882-w
citation_public_url	https://hdl.handle.net/11094/99697

Modulation of Ca²⁺ oscillation following ischemia and nicotinic acetylcholine receptors in primary cortical neurons by high-throughput analysis

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