{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T01:27:09Z","timestamp":1790731629219,"version":"4.1.0"},"reference-count":50,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,4,11]],"date-time":"2022-04-11T00:00:00Z","timestamp":1649635200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Neurosci."],"abstract":"<jats:p>Psychological stress is commonly accepted to be closely associated with masticatory muscle disorder, which is the main symptom of temporomandibular disorder (TMD). Previous studies have confirmed that exposure to stress may cause masticatory muscle hyperactivity. However, the central mechanism underlying this process remains unclear. The mesencephalic trigeminal nucleus (Vme), which resides in the brainstem, is the primary afferent center for masticatory proprioception and plays a key role in oral\u2013motor movements by projecting to the trigeminal motor nucleus (Vmo). Therefore, the present study was designed to examine the role of Vme neurons in masseter overactivity induced by chronic stress. We found that subjecting mice to restraint stress (6 h\/day) for 14 days caused significant anxiety-like behavior, obvious masseter overactivity, and markedly enhanced electrophysiological excitability of Vme neurons. By using anterograde tract tracing combined with immunofluorescence staining methods, we observed vesicular glutamate transporter 1 (VGLUT1)-positive glutamatergic projections from the Vme to the Vmo. Moreover, chronic restraint stress (CRS) elevated the expression of VGLUT1 and choline acetyltransferase (ChAT) in Vmo. Furthermore, administration of VGLUT1-targeted short hairpin RNA (shRNA) into the bilateral Vme significantly suppressed the enhanced overexcitability of Vme neurons, downregulated the overexpression of VGLUT1 and ChAT in the Vmo, and attenuated the elevated overactivity of the masseter caused by CRS. Taken together, we showed that CRS can excite neurons in the Vme, enhancing glutamatergic excitatory projections from the Vme to the Vmo and resulting in masseter muscle overactivity. These findings provide us with a novel central mechanism underlying the correlation between psychological factors and TMD.<\/jats:p>","DOI":"10.3389\/fncel.2022.841133","type":"journal-article","created":{"date-parts":[[2022,4,11]],"date-time":"2022-04-11T04:48:07Z","timestamp":1649652487000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":25,"title":["Activation of the Mesencephalic Trigeminal Nucleus Contributes to Masseter Hyperactivity Induced by Chronic Restraint Stress"],"prefix":"10.3389","volume":"16","author":[{"given":"Ya-Juan","family":"Zhao","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiang","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhou-Ming","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Teng","family":"Tu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rong","family":"Lei","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Min","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yong-Jin","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2022,4,11]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"700","DOI":"10.1038\/nn.4526","article-title":"C1 neurons mediate a stress-induced anti-inflammatory reflex in mice.","volume":"20","author":"Abe","year":"2017","journal-title":"Nat. Neurosci."},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1186\/s10194-016-0622-0","article-title":"The associations between psychosocial aspects and TMD-pain related aspects in children and adolescents.","volume":"17","author":"Al-Khotani","year":"2016","journal-title":"J. Headache Pain"},{"key":"B3","doi-asserted-by":"publisher","first-page":"39e","DOI":"10.1097\/prs.0000000000005083","article-title":"The masseter muscle and its role in facial contouring, aging, and quality of life: a literature review.","volume":"143","author":"Almukhtar","year":"2019","journal-title":"Plast. Reconstr. Surg."},{"key":"B4","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1111\/j.1601-0825.2011.01790.x","article-title":"Persistent orofacial muscle pain.","volume":"17","author":"Benoliel","year":"2011","journal-title":"Oral Dis."},{"key":"B5","doi-asserted-by":"publisher","first-page":"311","DOI":"10.1016\/j.ecoenv.2016.03.043","article-title":"Behavioral swimming effects and acetylcholinesterase activity changes in Jenynsia multidentata exposed to chlorpyrifos and cypermethrin individually and in mixtures.","volume":"129","author":"Bonansea","year":"2016","journal-title":"Ecotoxicol. Environ. Saf."},{"key":"B6","doi-asserted-by":"publisher","first-page":"209","DOI":"10.1093\/bmb\/ldm014","article-title":"Creatine kinase monitoring in sport medicine.","volume":"81","author":"Brancaccio","year":"2007","journal-title":"Br. Med. Bull."},{"key":"B7","doi-asserted-by":"publisher","first-page":"112","DOI":"10.1016\/j.pnpbp.2012.05.018","article-title":"Chronic restraint stress causes anxiety- and depression-like behaviors, downregulates glucocorticoid receptor expression, and attenuates glutamate release induced by brain-derived neurotrophic factor in the prefrontal cortex.","volume":"39","author":"Chiba","year":"2012","journal-title":"Prog. Neuropsychopharmacol. Biol. Psychiatry"},{"key":"B8","doi-asserted-by":"publisher","first-page":"640","DOI":"10.1111\/joor.12644","article-title":"Muscle hardness and masticatory myofascial pain: assessment and clinical relevance.","volume":"45","author":"Costa","year":"2018","journal-title":"J. Oral Rehabil."},{"key":"B9","doi-asserted-by":"publisher","first-page":"80","DOI":"10.3109\/00016350903431118","article-title":"Temporomandibular disorders and oral health-related quality of life. A systematic review.","volume":"68","author":"Dahlstr\u00f6m","year":"2010","journal-title":"Acta Odontol. Scand."},{"key":"B10","doi-asserted-by":"publisher","first-page":"T75","DOI":"10.1016\/j.jpain.2013.06.009","article-title":"Psychological factors associated with development of TMD: the OPPERA prospective cohort study.","volume":"14","author":"Fillingim","year":"2013","journal-title":"J. Pain"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.3390\/nu13051642","article-title":"The mesencephalic trigeminal nucleus controls food intake and body weight via hindbrain POMC projections.","volume":"13","author":"Fortin","year":"2021","journal-title":"Nutrients"},{"key":"B12","doi-asserted-by":"publisher","first-page":"261","DOI":"10.1007\/s00784-006-0064-1","article-title":"How may stressful experiences contribute to the development of temporomandibular disorders?","volume":"10","author":"Gameiro","year":"2006","journal-title":"Clin. Oral Investig."},{"key":"B13","doi-asserted-by":"publisher","first-page":"877","DOI":"10.1016\/s0022-2836(03)00921-5","article-title":"Muscle-type creatine kinase interacts with central domains of the M-band proteins myomesin and M-protein.","volume":"332","author":"Hornemann","year":"2003","journal-title":"J. Mol. Biol."},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2017.00489","article-title":"Microbiota modulate anxiety-like behavior and endocrine abnormalities in hypothalamic-pituitary-adrenal axis.","volume":"7","author":"Huo","year":"2017","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"B15","doi-asserted-by":"publisher","first-page":"1188","DOI":"10.1016\/j.jpain.2012.09.004","article-title":"Depressive and anxiety symptoms as risk factors for temporomandibular joint pain: a prospective cohort study in the general population.","volume":"13","author":"Kindler","year":"2012","journal-title":"J. Pain"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.2186\/jpr.JPR_D_21_00171","article-title":"Effect of masseter muscle activity during wakefulness and sleep on tooth wear.","author":"Kitagawa","year":"2021"},{"key":"B17","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1016\/s0301-0082(01)00021-1","article-title":"Comparative analysis of the chemical neuroanatomy of the mammalian trigeminal ganglion and mesencephalic trigeminal nucleus.","volume":"66","author":"Lazarov","year":"2002","journal-title":"Prog. Neurobiol."},{"key":"B18","doi-asserted-by":"publisher","first-page":"362","DOI":"10.1016\/j.brainresrev.2007.08.009","article-title":"Neurobiology of orofacial proprioception.","volume":"56","author":"Lazarov","year":"2007","journal-title":"Brain Res. Rev."},{"key":"B19","doi-asserted-by":"publisher","first-page":"118","DOI":"10.1016\/j.aanat.2010.12.004","article-title":"Amygdalotrigeminal projection in the rat: an anterograde tracing study.","volume":"193","author":"Lazarov","year":"2011","journal-title":"Ann. Anat."},{"key":"B20","doi-asserted-by":"publisher","first-page":"202","DOI":"10.1016\/j.expneurol.2019.03.009","article-title":"Muscle ciliary neurotrophic factor receptor \u03b1 helps maintain choline acetyltransferase levels in denervated motor neurons following peripheral nerve lesion.","volume":"317","author":"Lee","year":"2019","journal-title":"Exp. Neurol."},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.3389\/fncel.2019.00495","article-title":"NMDAR and JNK activation in the spinal trigeminal nucleus caudalis contributes to masseter hyperalgesia induced by stress.","volume":"13","author":"Lin","year":"2019","journal-title":"Front. Cell. Neurosci."},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1186\/s13064-017-0088-z","article-title":"The emergence of mesencephalic trigeminal neurons.","volume":"12","author":"Lipovsek","year":"2017","journal-title":"Neural Dev."},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-15920-7","article-title":"Identification of a prefrontal cortex-to-amygdala pathway for chronic stress-induced anxiety.","volume":"11","author":"Liu","year":"2020","journal-title":"Nat. Commun."},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.3389\/fnmol.2019.00174","article-title":"Malocclusion generates anxiety-like behavior through a putative lateral habenula-mesencephalic trigeminal nucleus pathway.","volume":"12","author":"Liu","year":"2019","journal-title":"Front. Mol. Neurosci."},{"key":"B25","doi-asserted-by":"publisher","first-page":"941","DOI":"10.1007\/s00421-011-1961-9","article-title":"Creatine kinase MM TaqI and methylenetetrahydrofolate reductase C677T and A1298C gene polymorphisms influence exercise-induced C-reactive protein levels.","volume":"112","author":"Miranda-Vilela","year":"2012","journal-title":"Eur. J. Appl. Physiol."},{"key":"B26","doi-asserted-by":"publisher","first-page":"514","DOI":"10.14219\/jada.archive.1988.0320","article-title":"Masticatory muscle hyperactivity in temporomandibular disorders: is it an extrapyramidally expressed disorder?","volume":"116","author":"Nishioka","year":"1988","journal-title":"J. Am. Dent. Assoc."},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.3389\/fneur.2019.00250","article-title":"Platelet-Rich Plasma Intramuscular Injections - Antinociceptive Therapy in Myofascial Pain Within Masseter Muscles in Temporomandibular Disorders Patients: a Pilot Study.","volume":"10","author":"Nitecka-Buchta","year":"2019","journal-title":"Front. Neurol."},{"key":"B28","doi-asserted-by":"publisher","first-page":"1052","DOI":"10.1111\/joor.12994","article-title":"The impact of oro-facial pain conditions on oral health-related quality of life: a systematic review.","volume":"47","author":"Oghli","year":"2020","journal-title":"J. Oral Rehabil."},{"key":"B29","doi-asserted-by":"publisher","first-page":"304","DOI":"10.1080\/08869634.2017.1361052","article-title":"Frequency of temporomandibular disorders diagnoses based on RDC\/TMD in a Polish patient population.","volume":"36","author":"Osiewicz","year":"2018","journal-title":"Cranio"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1177\/0300060519889487","article-title":"Manifestation of stress and anxiety in the stomatognathic system of undergraduate dentistry students.","volume":"48","author":"Owczarek","year":"2020","journal-title":"J. Int. Med. Res."},{"key":"B31","doi-asserted-by":"publisher","first-page":"595","DOI":"10.1002\/cne.21894","article-title":"Axon terminals expressing vesicular glutamate transporter VGLUT1 or VGLUT2 within the trigeminal motor nucleus of the rat: origins and distribution patterns.","volume":"512","author":"Pang","year":"2009","journal-title":"J. Comp. Neurol."},{"key":"B32","doi-asserted-by":"publisher","first-page":"129","DOI":"10.1002\/cne.21047","article-title":"Expression of vesicular glutamate transporter 1 immunoreactivity in peripheral and central endings of trigeminal mesencephalic nucleus neurons in the rat.","volume":"498","author":"Pang","year":"2006","journal-title":"J. Comp. Neurol."},{"key":"B33","doi-asserted-by":"publisher","first-page":"2323","DOI":"10.1007\/s00429-018-1636-y","article-title":"Vesicular glutamate transporter 1 (VGLUT1)- and VGLUT2-immunopositive axon terminals on the rat jaw-closing and jaw-opening motoneurons.","volume":"223","author":"Park","year":"2018","journal-title":"Brain Struct. Funct."},{"key":"B34","author":"Paxinos","year":"2001","journal-title":"The Mouse Brain in Stereotaxic Coordinates."},{"key":"B35","doi-asserted-by":"publisher","first-page":"1106","DOI":"10.1523\/jneurosci.4182-06.2007","article-title":"The role of muscle activation pattern and calcineurin in acetylcholinesterase regulation in rat skeletal muscles.","volume":"27","author":"Pregelj","year":"2007","journal-title":"J. Neurosci."},{"key":"B36","doi-asserted-by":"publisher","first-page":"3197","DOI":"10.1007\/s00784-020-03650-5","article-title":"The intensity of awake bruxism episodes is increased in individuals with high trait anxiety.","volume":"25","author":"Rofaeel","year":"2021","journal-title":"Clin. Oral Investig."},{"key":"B37","doi-asserted-by":"publisher","first-page":"107","DOI":"10.1093\/ejo\/24.1.107","article-title":"Emotional stress and brux-like activity of the masseter muscle in rats.","volume":"24","author":"Rosales","year":"2002","journal-title":"Eur. J. Orthod."},{"key":"B38","doi-asserted-by":"publisher","first-page":"247","DOI":"10.1007\/bf02936610","article-title":"Molecular biology and neurobiology of choline acetyltransferase.","volume":"1","author":"Salvaterra","year":"1987","journal-title":"Mol. Neurobiol."},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.3389\/fncel.2018.00497","article-title":"Increased anxiety-related behavior, impaired cognitive function and cellular alterations in the brain of Cend1-deficient mice.","volume":"12","author":"Segklia","year":"2019","journal-title":"Front. Cell. Neurosci."},{"key":"B40","doi-asserted-by":"publisher","first-page":"666","DOI":"10.1177\/0022034519837249","article-title":"Masseter Deoxygenation in Adults at Risk for Temporomandibular Disorders.","volume":"98","author":"Shah","year":"2019","journal-title":"J. Dent. Res."},{"key":"B41","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1016\/j.brainres.2011.05.026","article-title":"Direct projections from the central amygdaloid nucleus to the mesencephalic trigeminal nucleus in rats.","volume":"1400","author":"Shirasu","year":"2011","journal-title":"Brain Res."},{"key":"B42","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1016\/j.cbi.2008.05.035","article-title":"Acetylcholinesterase: how is structure related to function?","volume":"175","author":"Silman","year":"2008","journal-title":"Chem. Biol. Interact."},{"key":"B43","doi-asserted-by":"publisher","first-page":"3939","DOI":"10.1007\/s00784-020-03260-1","article-title":"Depression and anxiety levels in patients with temporomandibular disorders: comparison with the general population.","volume":"24","author":"Simoen","year":"2020","journal-title":"Clin. Oral Investig."},{"key":"B44","doi-asserted-by":"publisher","first-page":"1084","DOI":"10.1177\/0022034516653743","article-title":"Painful temporomandibular disorder: decade of discovery from OPPERA studies.","volume":"95","author":"Slade","year":"2016","journal-title":"J. Dent. Res."},{"key":"B45","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1016\/j.physbeh.2014.06.017","article-title":"Lamotrigine reverses masseter overactivity caused by stress maybe via Glu suppression.","volume":"137","author":"Song","year":"2014","journal-title":"Physiol. Behav."},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.57515","article-title":"Acetylcholine in action.","volume":"9","author":"Wall","year":"2020","journal-title":"Elife"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.3389\/fncel.2015.00493","article-title":"Corticotrigeminal projections from the insular cortex to the trigeminal caudal subnucleus regulate orofacial pain after nerve injury via extracellular signal-regulated kinase activation in insular cortex neurons.","volume":"9","author":"Wang","year":"2015","journal-title":"Front. Cell. Neurosci."},{"key":"B48","doi-asserted-by":"publisher","first-page":"1077","DOI":"10.1111\/joor.13033","article-title":"Inter-relationships between pain-related temporomandibular disorders, somatic and psychological symptoms in Asian youths.","volume":"47","author":"Yap","year":"2020","journal-title":"J. Oral Rehabil."},{"key":"B49","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1016\/j.brainres.2012.10.021","article-title":"Jaw muscle spindle afferents coordinate multiple orofacial motoneurons via common premotor neurons in rats: an electrophysiological and anatomical study.","volume":"1489","author":"Zhang","year":"2012","journal-title":"Brain Res."},{"key":"B50","doi-asserted-by":"publisher","first-page":"109","DOI":"10.1016\/0304-3959(83)90201-4","article-title":"Ethical guidelines for investigations of experimental pain in conscious animals.","volume":"16","author":"Zimmermann","year":"1983","journal-title":"Pain"}],"container-title":["Frontiers in Cellular Neuroscience"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fncel.2022.841133\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,4,11]],"date-time":"2022-04-11T04:48:11Z","timestamp":1649652491000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fncel.2022.841133\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,4,11]]},"references-count":50,"alternative-id":["10.3389\/fncel.2022.841133"],"URL":"https:\/\/doi.org\/10.3389\/fncel.2022.841133","relation":{},"ISSN":["1662-5102"],"issn-type":[{"value":"1662-5102","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,4,11]]},"article-number":"841133"}}