{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T00:16:15Z","timestamp":1788480975029,"version":"build-2803163510"},"reference-count":31,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2022,9,28]],"date-time":"2022-09-28T00:00:00Z","timestamp":1664323200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Natural Science Foundation of China","award":["82071147"],"award-info":[{"award-number":["82071147"]}]},{"name":"National Natural Science Foundation of China","award":["82171000"],"award-info":[{"award-number":["82171000"]}]},{"name":"National Natural Science Foundation of China","award":["2021YJ0428"],"award-info":[{"award-number":["2021YJ0428"]}]},{"name":"Sichuan Science and Technology Program","award":["82071147"],"award-info":[{"award-number":["82071147"]}]},{"name":"Sichuan Science and Technology Program","award":["82171000"],"award-info":[{"award-number":["82171000"]}]},{"name":"Sichuan Science and Technology Program","award":["2021YJ0428"],"award-info":[{"award-number":["2021YJ0428"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The nerve growth factor (NGF) and calcitonin gene-related peptide (CGRP) play a crucial role in the regulation of orofacial pain. It has been demonstrated that CGRP increases orofacial pain induced by NGF. V-type proton ATPase subunit an isoform 1 (Atp6v0a1) is involved in the exocytosis pathway, especially in vesicular transport in neurons. The objective was to examine the role of Atp6v0a1 in NGF-induced upregulation of CGRP in orofacial pain induced by experimental tooth movement. Orofacial pain was elicited by ligating closed-coil springs between incisors and molars in Sprague\u2013Dawley rats. Gene and protein expression levels were determined through real-time polymerase chain reaction, immunostaining, and fluorescence in situ hybridization. Lentivirus vectors carrying Atp6v0a1 shRNA were used to knockdown the expression of Atp6v0a1 in TG and SH-SY5Y neurons. The release of vesicles in SH-SY5Y neurons was observed by using fluorescence dye FM1-43, and the release of CGRP was detected by Enzyme-Linked Immunosorbent Assy. Orofacial pain was evaluated through the rat grimace scale. Our results revealed that intraganglionic administration of NGF and Atp6v0a1 shRNA upregulated and downregulated CGRP in trigeminal ganglia (TG) and trigeminal subnucleus caudalis (Vc), respectively, and the orofacial pain was also exacerbated and alleviated, respectively, following administration of NGF and Atp6v0a1 shRNA. Besides, intraganglionic administration of NGF simultaneously caused the downregulation of Atp6v0a1 in TG. Moreover, the release of vesicles and CGRP in SH-SY5Y neurons was interfered by NGF and Atp6v0a1 shRNA. In conclusion, in the orofacial pain induced by experimental tooth movement, NGF induced the upregulation of CGRP in TG and Vc, and this process is dependent on Atp6v0a1 and vesicle release, suggesting that they are involved in the transmission of nociceptive information in orofacial pain.<\/jats:p>","DOI":"10.3390\/ijms231911440","type":"journal-article","created":{"date-parts":[[2022,9,28]],"date-time":"2022-09-28T22:53:19Z","timestamp":1664405599000},"page":"11440","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["NGF-Induced Upregulation of CGRP in Orofacial Pain Induced by Tooth Movement Is Dependent on Atp6v0a1 and Vesicle Release"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4944-6591","authenticated-orcid":false,"given":"Tianjin","family":"Tao","sequence":"first","affiliation":[{"name":"State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral Diseases, Sichuan University, Chengdu 610041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi","family":"Liu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral Diseases, Sichuan University, Chengdu 610041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingqi","family":"Zhang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral Diseases, Sichuan University, Chengdu 610041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenli","family":"Lai","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral Diseases, Sichuan University, Chengdu 610041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hu","family":"Long","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral Diseases, Sichuan University, Chengdu 610041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,9,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"721","DOI":"10.1016\/j.adaj.2015.04.001","article-title":"Prevalence of pain in the orofacial regions in patients visiting general dentists in the Northwest Practice-based REsearch Collaborative in Evidence-based DENTistry research network","volume":"146","author":"Horst","year":"2015","journal-title":"J. Am. Dent. Assoc."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1177\/0022034520950296","article-title":"CGRP Modulates Orofacial Pain through Mediating Neuron-Glia Crosstalk","volume":"100","author":"Liang","year":"2021","journal-title":"J. Dent. Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"k1517","DOI":"10.1136\/bmj.k1517","article-title":"Orofacial pain","volume":"361","author":"Banigo","year":"2018","journal-title":"BMJ"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1111\/ger.12063","article-title":"Prevalence of headache and orofacial pain in adults and elders in a Brazilian community: An epidemiological study","volume":"32","author":"Vilela","year":"2015","journal-title":"Gerodontology"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"492","DOI":"10.2174\/1874210601711010492","article-title":"Methodologies in Orthodontic Pain Management: A Review","volume":"11","author":"Hussain","year":"2017","journal-title":"Open Dent. J."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1038\/s41368-019-0044-3","article-title":"Transient receptor potential Vanilloid 1-based gene therapy alleviates orthodontic pain in rats","volume":"11","author":"Guo","year":"2019","journal-title":"Int. J. Oral Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"104817","DOI":"10.1016\/j.archoralbio.2020.104817","article-title":"Botulinum toxin A alleviates orofacial nociception induced by orthodontic tooth movement through nociceptin\/orphanin-FQ pathway in rats","volume":"117","author":"Lyu","year":"2020","journal-title":"Arch. Oral Biol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1038\/ijos.2016.24","article-title":"Current advances in orthodontic pain","volume":"8","author":"Long","year":"2016","journal-title":"Int. J. Oral Sci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1875","DOI":"10.2147\/JPR.S250030","article-title":"Role of Nerve Growth Factor in Orofacial Pain","volume":"13","author":"Mai","year":"2020","journal-title":"J. Pain Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1038\/s41584-020-00528-4","article-title":"The evolution of nerve growth factor inhibition in clinical medicine","volume":"17","author":"Wise","year":"2021","journal-title":"Nat. Rev. Rheumatol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1038\/s41368-021-00124-6","article-title":"Retrograde nerve growth factor signaling modulates tooth mechanical hyperalgesia induced by orthodontic tooth movement via acid-sensing ion channel 3","volume":"13","author":"Gao","year":"2021","journal-title":"Int. J. Oral Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"385","DOI":"10.3389\/fnins.2020.00385","article-title":"Spinal Mobilization Prevents NGF-Induced Trunk Mechanical Hyperalgesia and Attenuates Expression of CGRP","volume":"14","author":"Reed","year":"2020","journal-title":"Front. Neurosci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"597","DOI":"10.1590\/1678-775720160150","article-title":"The effect of capsaicin on expression patterns of CGRP in trigeminal ganglion and trigeminal nucleus caudalis following experimental tooth movement in rats","volume":"24","author":"Zhou","year":"2016","journal-title":"J. Appl. Oral Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"7a","DOI":"10.1016\/j.bpj.2015.11.086","article-title":"How to Fill a Synaptic Vesicle with Neurotransmitters? Transport Mechanisms and Ion Balance","volume":"110","author":"Jahn","year":"2016","journal-title":"Biophys. J."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1649","DOI":"10.7150\/thno.40919","article-title":"Nerve growth factor activates autophagy in Schwann cells to enhance myelin debris clearance and to expedite nerve regeneration","volume":"10","author":"Li","year":"2020","journal-title":"Theranostics"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1619","DOI":"10.1002\/ejp.1438","article-title":"Masseter corticomotor excitability is decreased after intramuscular administration of nerve growth factor","volume":"23","author":"Costa","year":"2019","journal-title":"Eur. J. Pain"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1007\/978-3-662-46450-2_4","article-title":"Role of nerve growth factor in pain","volume":"227","author":"Mizumura","year":"2015","journal-title":"Handb. Exp. Pharmacol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"7145","DOI":"10.1523\/JNEUROSCI.0481-11.2011","article-title":"Nerve growth factor contribution via transient receptor potential vanilloid 1 to ectopic orofacial pain","volume":"31","author":"Shinoda","year":"2011","journal-title":"J. Neurosci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1016\/j.neuroscience.2016.10.064","article-title":"The TrkA receptor mediates experimental thermal hyperalgesia produced by nerve growth factor: Modulation by the p75 neurotrophin receptor","volume":"340","author":"Khodorova","year":"2017","journal-title":"Neuroscience"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1093\/ejo\/cjv082","article-title":"The role of periodontal ASIC3 in orofacial pain induced by experimental tooth movement in rats","volume":"38","author":"Gao","year":"2016","journal-title":"Eur. J. Orthodont."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"993","DOI":"10.1016\/j.molcel.2018.02.006","article-title":"The 3.5-A degrees CryoEM Structure of Nanodisc-Reconstituted Yeast Vacuolar ATPase V-o Proton Channel","volume":"69","author":"Roh","year":"2018","journal-title":"Mol. Cell"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1186\/s12943-018-0811-3","article-title":"The curious case of vacuolar ATPase: Regulation of signaling pathways","volume":"17","author":"Pamarthy","year":"2018","journal-title":"Mol. Cancer"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2107","DOI":"10.1038\/s41467-021-22389-5","article-title":"ATP6V0A1 encoding the a1-subunit of the V0 domain of vacuolar H(+)-ATPases is essential for brain development in humans and mice","volume":"12","author":"Aoto","year":"2021","journal-title":"Nat. Commun."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"3394","DOI":"10.1091\/mbc.e11-02-0155","article-title":"Vacuolar H(+)-ATPase subunits Voa1 and Voa2 cooperatively regulate secretory vesicle acidification, transmitter uptake, and storage","volume":"22","author":"Saw","year":"2011","journal-title":"Mol. Biol. Cell"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"105001","DOI":"10.1016\/j.neuint.2021.105001","article-title":"HAGLR aggravates neuropathic pain and promotes inflammatory response and apoptosis of lipopolysaccharide-treated SH-SY5Y cells by sequestering miR-182-5p from ATAT1 and activating NLRP3 inflammasome","volume":"145","author":"Zhang","year":"2021","journal-title":"Neurochem. Int."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1469","DOI":"10.1007\/s12011-020-02250-4","article-title":"Melatonin and Selenium Suppress Docetaxel-Induced TRPV1 Activation, Neuropathic Pain and Oxidative Neurotoxicity in Mice","volume":"199","author":"Ertilav","year":"2021","journal-title":"Biol. Trace Elem. Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"92","DOI":"10.3389\/fphar.2020.00092","article-title":"Zerumbone Modulates alpha2A-Adrenergic, TRPV1, and NMDA NR2B Receptors Plasticity in CCI-Induced Neuropathic Pain In Vivo and LPS-Induced SH-SY5Y Neuroblastoma In Vitro Models","volume":"11","author":"Chia","year":"2020","journal-title":"Front. Pharmacol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1111\/eos.12110","article-title":"Evaluation of pain in rats through facial expression following experimental tooth movement","volume":"122","author":"Liao","year":"2014","journal-title":"Eur. J. Oral Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1111\/eos.12326","article-title":"A novel technique of delivering viral vectors to trigeminal ganglia in rats","volume":"125","author":"Long","year":"2017","journal-title":"Eur. J. Oral Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"7297","DOI":"10.1038\/s41598-018-25453-1","article-title":"The contribution of microglia to early synaptic compensatory responses that precede beta-amyloid-induced neuronal death","volume":"8","author":"Merlo","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"4229491","DOI":"10.1155\/2021\/4229491","article-title":"Mechanism of Catechol-O-methyltransferase Regulating Orofacial Pain Induced by Tooth Movement","volume":"2021","author":"Zhou","year":"2021","journal-title":"BioMed Res. Int."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/19\/11440\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:41:01Z","timestamp":1760143261000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/19\/11440"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,9,28]]},"references-count":31,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2022,10]]}},"alternative-id":["ijms231911440"],"URL":"https:\/\/doi.org\/10.3390\/ijms231911440","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,9,28]]}}}