PMID- 37955123 OWN - NLM STAT- MEDLINE DCOM- 20240109 LR - 20240109 IS - 1522-1563 (Electronic) IS - 0363-6143 (Linking) VI - 326 IP - 1 DP - 2024 Jan 1 TI - Caveolin-1 forms a complex with P2X7 receptor and tunes P2X7-mediated ATP signaling in mouse bone marrow-derived macrophages. PG - C125-C142 LID - 10.1152/ajpcell.00303.2023 [doi] AB - The ionotropic purinergic P2X7 receptor responds to extracellular ATP and can trigger proinflammatory immune signaling in macrophages. Caveolin-1 (Cav-1) is known to modulate functions of macrophages and innate immunity. However, it is unknown how Cav-1 modulates P2X7 receptor activity in macrophages. We herein examined P2X7 receptor activity and macrophage functions using bone marrow-derived macrophages (BMDMs) from wild-type (WT) and Cav-1 knockout (KO) mice. ATP (1 mM) application caused biphasic increase in cytosolic [Ca(2+)] and sustained decrease in cytosolic [K(+)]. A specific P2X7 receptor blocker, A-740003, inhibited the maintained cytosolic [Ca(2+)] increase and cytosolic [K(+)] decrease. Total internal reflection fluorescent imaging and proximity ligation assays revealed a novel molecular complex formation between P2X7 receptors and Cav-1 in WT BMDMs that were stimulated with lipopolysaccharides. This molecular coupling was increased by ATP application. Specifically, the ATP-induced Ca(2+) influx and K(+) efflux through P2X7 receptors were increased in Cav-1 KO BMDMs, even though the total and surface protein levels of P2X7 receptors in WT and Cav-1 KO BMDMs were unchanged. Cell-impermeable dye (TO-PRO3) uptake analysis revealed that macropore formation of P2X7 receptors was enhanced in Cav-1 KO BMDMs. Cav-1 KO BMDMs increased ATP-induced IL-1beta secretion, reactive oxygen species production, Gasdermin D (GSDMD) cleavage, and lactate dehydrogenase release indicating pyroptosis. A-740003 completely prevented ATP-induced pyroptosis. In combination, these datasets show that Cav-1 has a negative effect on P2X7 receptor activity in BMDMs and that Cav-1 in macrophages may contribute to finely tuned immune responses by preventing excessive IL-1beta secretion and pyroptosis.NEW & NOTEWORTHY In bone marrow-derived macrophages, Cav-1 suppresses the macropore formation of P2X7 receptors through their direct or indirect interactions, resulting in reduced membrane permeability of cations (Ca(2+) and K(+)) and large cell-impermeable dye (TO-PRO3) induced by ATP. Cav-1 also inhibits ATP-induced IL-1beta secretion, ROS production, GSDMD cleavage, and pyroptosis. Cav-1 contributes to the maintenance of proper immune responses by finely tuning IL-1beta secretion and cell death in macrophages. FAU - Sawai, Yuuki AU - Sawai Y AD - Department of Molecular and Cellular Pharmacology, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan. ROR: https://ror.org/04wn7wc95 FAU - Suzuki, Yoshiaki AU - Suzuki Y AUID- ORCID: 0000-0002-7879-6059 AD - Department of Molecular and Cellular Pharmacology, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan. ROR: https://ror.org/04wn7wc95 FAU - Asagiri, Masataka AU - Asagiri M AD - Department of Pharmacology, Yamaguchi University Graduate School of Medicine, Ube, Japan. FAU - Hida, Shigeaki AU - Hida S AD - Department of Molecular and Cellular Health Sciences, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan. FAU - Kondo, Rubii AU - Kondo R AD - Department of Molecular and Cellular Pharmacology, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan. ROR: https://ror.org/04wn7wc95 FAU - Zamponi, Gerald W AU - Zamponi GW AD - Department of Physiology and Pharmacology, Cumming School of Medicine, University of Calgary, Calgary, Alberta, Canada. AD - Department of Clinical Neurosciences, Hotchkiss Brain Institute, Alberta Children's Hospital Research Institute, Cumming School of Medicine, University of Calgary, Calgary, Alberta, Canada. FAU - Giles, Wayne R AU - Giles WR AD - Department of Physiology and Pharmacology, Cumming School of Medicine, University of Calgary, Calgary, Alberta, Canada. FAU - Imaizumi, Yuji AU - Imaizumi Y AUID- ORCID: 0000-0001-9431-688X AD - Department of Molecular and Cellular Pharmacology, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan. ROR: https://ror.org/04wn7wc95 FAU - Yamamura, Hisao AU - Yamamura H AUID- ORCID: 0000-0001-8909-176X AD - Department of Molecular and Cellular Pharmacology, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan. ROR: https://ror.org/04wn7wc95 LA - eng GR - 22H02773/MEXT | Japan Society for the Promotion of Science (JSPS)/ GR - 21K19343/MEXT | Japan Society for the Promotion of Science (JSPS)/ GR - 19H03381/MEXT | Japan Society for the Promotion of Science (JSPS)/ GR - 16H06215/MEXT | Japan Society for the Promotion of Science (JSPS)/ GR - 16K15127/MEXT | Japan Society for the Promotion of Science (JSPS)/ GR - 22H02787/MEXT | Japan Society for the Promotion of Science (JSPS)/ GR - 19K07125/MEXT | Japan Society for the Promotion of Science (JSPS)/ GR - 18KK0218/MEXT | Japan Society for the Promotion of Science (JSPS)/ GR - N/A/Pharmacological Research Foundation/ GR - N/A/Suzuken Memorial Foundation/ GR - N/A/Japan Foundation for Applied Enzymology/ GR - N/A/Canada Research Chairs (Chaires de recherche du Canada)/ GR - N/A/Takeda Science Foundation (TSF)/ PT - Journal Article DEP - 20231113 PL - United States TA - Am J Physiol Cell Physiol JT - American journal of physiology. Cell physiology JID - 100901225 RN - 0 ((N-(1-(((cyanoimino)(5-quinolinylamino) methyl) amino)-2,2-dimethylpropyl)-2-(3,4-dimethoxyphenyl)acetamide)) RN - 8L70Q75FXE (Adenosine Triphosphate) RN - 0 (Caveolin 1) RN - 0 (Interleukin-1beta) RN - 0 (Receptors, Purinergic P2X7) RN - 0 (Cav1 protein, mouse) RN - 0 (P2rx7 protein, mouse) SB - IM MH - Animals MH - Mice MH - Adenosine Triphosphate/pharmacology/metabolism MH - *Caveolin 1/genetics/metabolism MH - Interleukin-1beta/metabolism MH - Macrophages/metabolism MH - *Receptors, Purinergic P2X7/metabolism OTO - NOTNLM OT - ATP OT - P2X7 OT - calcium channel OT - caveolin-1 OT - macrophage EDAT- 2023/11/13 06:43 MHDA- 2024/01/03 09:43 CRDT- 2023/11/13 05:27 PHST- 2024/01/03 09:43 [medline] PHST- 2023/11/13 06:43 [pubmed] PHST- 2023/11/13 05:27 [entrez] AID - 10.1152/ajpcell.00303.2023 [doi] PST - ppublish SO - Am J Physiol Cell Physiol. 2024 Jan 1;326(1):C125-C142. doi: 10.1152/ajpcell.00303.2023. Epub 2023 Nov 13.