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The underlying mechanisms that downregulate osteocytes\u2019 sensitivity and overall response to loading remain unknown. In other cell types, purinergic signaling through the P2Y2 receptor has the potential to downregulate the sensitivity to loading by modifying cell stiffness through actin polymerization and cytoskeleton organization. Herein, we examined the role of P2Y2 activation in regulating osteocytes\u2019 mechanotransduction using a P2Y2 knockout cell line alongside conditional knockout mice. Our findings demonstrate that the absence of P2Y2 expression in MLO-Y4 cells prevents actin polymerization while increasing the sensitivity to fluid flow\u2013induced shear stress. Deleting osteocytes\u2019 P2Y2 expression in conditional-knockout mice enabled bone formation to increase when increasing the duration of exercise. Overall, P2Y2 activation under loading produces a negative feedback loop, limiting osteocytes\u2019 response to continuous loading by shifting the sensitivity to mechanical strain through actin stress fiber formation.<\/jats:p>","DOI":"10.1083\/jcb.202403005","type":"journal-article","created":{"date-parts":[[2024,8,8]],"date-time":"2024-08-08T02:21:19Z","timestamp":1723083679000},"update-policy":"https:\/\/doi.org\/10.1083\/jcb.crossmarkpolicy","source":"Crossref","is-referenced-by-count":8,"title":["Purinergic signaling through the P2Y2 receptor regulates osteocytes\u2019 mechanosensitivity"],"prefix":"10.1083","volume":"223","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2981-3745","authenticated-orcid":true,"given":"Amit","family":"Chougule","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02kwnkm68","id-type":"ROR","asserted-by":"publisher"}],"name":"Bone and Joint Center, Henry Ford Health System 1 , Detroit, MI,","place":["USA"]},{"id":[{"id":"https:\/\/ror.org\/05hs6h993","id-type":"ROR","asserted-by":"publisher"}],"name":"Henry Ford Health + Michigan State University Health Sciences 2 , Detroit, MI,","place":["USA"]},{"id":[{"id":"https:\/\/ror.org\/05hs6h993","id-type":"ROR","asserted-by":"publisher"}],"name":"College of Human Medicine, Michigan State University 3 Department Physiology, , East Lansing, MI,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-6477-2453","authenticated-orcid":true,"given":"Chunbin","family":"Zhang","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02kwnkm68","id-type":"ROR","asserted-by":"publisher"}],"name":"Bone and Joint Center, Henry Ford Health System 1 , Detroit, MI,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-8156-3350","authenticated-orcid":true,"given":"Nickolas","family":"Vinokurov","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01070mq45","id-type":"ROR","asserted-by":"publisher"}],"name":"School of Medicine, Wayne State University 4 , Detroit, MI,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-3455-9594","authenticated-orcid":true,"given":"Devin","family":"Mendez","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01070mq45","id-type":"ROR","asserted-by":"publisher"}],"name":"School of Medicine, Wayne State University 4 , Detroit, MI,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-3363-7540","authenticated-orcid":true,"given":"Elizabeth","family":"Vojtisek","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02kwnkm68","id-type":"ROR","asserted-by":"publisher"}],"name":"Bone and Joint Center, Henry Ford Health System 1 , Detroit, MI,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9581-7974","authenticated-orcid":true,"given":"Chenjun","family":"Shi","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01070mq45","id-type":"ROR","asserted-by":"publisher"}],"name":"Wayne State University 5 Department Biomedical Engineering, , Detroit, MI,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6240-0371","authenticated-orcid":true,"given":"Jitao","family":"Zhang","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01070mq45","id-type":"ROR","asserted-by":"publisher"}],"name":"Wayne State University 5 Department Biomedical Engineering, , Detroit, MI,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8651-8913","authenticated-orcid":true,"given":"Joseph","family":"Gardinier","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02kwnkm68","id-type":"ROR","asserted-by":"publisher"}],"name":"Bone and Joint Center, Henry Ford Health System 1 , Detroit, MI,","place":["USA"]},{"id":[{"id":"https:\/\/ror.org\/05hs6h993","id-type":"ROR","asserted-by":"publisher"}],"name":"Henry Ford Health + Michigan State University Health Sciences 2 , Detroit, MI,","place":["USA"]},{"id":[{"id":"https:\/\/ror.org\/05hs6h993","id-type":"ROR","asserted-by":"publisher"}],"name":"College of Human Medicine, Michigan State University 3 Department Physiology, , East Lansing, MI,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"291","published-online":{"date-parts":[[2024,8,30]]},"reference":[{"key":"2025082920015240400_bib1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.jmbbm.2011.08.026","article-title":"Overexpression of DMP1 accelerates mineralization and alters cortical bone biomechanical properties in vivo","volume":"5","author":"Bhatia","year":"2012","journal-title":"J. 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