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

Number of Access:1,081（2025-08-14 18:26 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/99697

Link to primary information

File	Format	Terms of use	Size	Views	Date.Available	Description	information
SciRep_14_27667	pdf	None	2.64 MB	35	2025.01.20

Creative Commons : Attribution - NonCommercial - NoDerivatives

Item Information

Output File Export EndNote Basic Export Mendeley

Title	Modulation of Ca²⁺ oscillation following ischemia and nicotinic acetylcholine receptors in primary cortical neurons by high-throughput analysis
Creator	NRID 1000020534879 1000020534879 ORCID 0000-0001-8726-978X 0000-0001-8726-978X researchmap permalink tsutomusasaki1111 tsutomusasaki1111 Scopus Author ID 7407394272 7407394272 CreatorName Sasaki, Tsutomu
	CreatorName Hisada, Sunao
	NRID 1000010774218 1000010774218 researchmap permalink kan1 kan1 CreatorName Kanki, Hideaki
	ORCID 0000-0002-6444-3244 0000-0002-6444-3244 researchmap permalink nunomurakazuto nunomurakazuto CreatorName Nunomura, Kazuto
	ORCID 0000-0001-5877-949X 0000-0001-5877-949X researchmap permalink linbangzhong linbangzhong Scopus Author ID 57200092216 57200092216 CreatorName Lin, Bangzhong
	CreatorName Nishiyama, Kumiko
	CreatorName Kawano, Tomohito
	NRID 1000070467413 1000070467413 CreatorName Matsumura, Shigenobu
	NRID 1000090230044 1000090230044 ORCID 0000-0002-0874-7542 0000-0002-0874-7542 researchmap permalink read0207787 read0207787 Scopus Author ID 7202006099 7202006099 CreatorName Mochizuki, Hideki
Subject	Ca²⁺ oscillations
	High-throughput screening
	Ischemia
	nAChR
	PAM
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.
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.
Publisher	Nature Research
Source Title	Scientific Reports
Volume (Issue)	14
Page	27667
Date of Issued	2024-11-12
Language	English
Handle URL	https://hdl.handle.net/11094/99697
eISSN	20452322
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1038/s41598-024-77882-w
Access Rights	open access
CopyRight	This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
oaire:version	Version of Record
Category	学術雑誌論文 Journal Article

Resource Type	journal article
Local Type	Journal Article
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	English
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	English
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

Link to primary information

Item Information

Show details