{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,12]],"date-time":"2026-05-12T18:21:13Z","timestamp":1778610073063,"version":"3.51.4"},"reference-count":52,"publisher":"MDPI AG","issue":"16","license":[{"start":{"date-parts":[[2020,8,11]],"date-time":"2020-08-11T00:00:00Z","timestamp":1597104000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000072","name":"National Institute of Dental and Craniofacial Research","doi-asserted-by":"publisher","award":["DE026749"],"award-info":[{"award-number":["DE026749"]}],"id":[{"id":"10.13039\/100000072","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000065","name":"National Institute of Neurological Disorders and Stroke","doi-asserted-by":"publisher","award":["NS064022"],"award-info":[{"award-number":["NS064022"]}],"id":[{"id":"10.13039\/100000065","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000053","name":"National Eye Institute","doi-asserted-by":"publisher","award":["EY08098"],"award-info":[{"award-number":["EY08098"]}],"id":[{"id":"10.13039\/100000053","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The orofacial pain pathway projects to the parabrachial and amygdala, and sex steroids have been shown to affect neuronal activity in these regions. GABA positive cells in the amygdala are influenced by sex steroid metabolites to affect pain, and sex steroids have been shown to alter the expression of genes in the parabrachial, changing neuronal excitability. Mechanisms by which sex steroids affect amygdala and parabrachial signaling are unclear. The expression of genes in the parabrachial and amygdala in diestrus (low estradiol) and proestrus (high estradiol) female rats were evaluated in this study. First, varicella zoster virus was injected into the whisker pad of female rats to induce a pain response. Second, gene expression was quantitated using RNA-seq one week after injection. Genes that had the greatest change in expression and known to function in pain signaling were selected for the quantitation of protein content. Protein expression of four genes in the parabrachial and seven genes in the amygdala were quantitated by ELISA. In the parabrachial, neurexin 3 (Nrnx3) was elevated at proestrus. Nrnx3 has a role in AMPA receptor and GABA signaling. Neuronatin (Nnat) and protein phosphatase, Mg2+\/Mn2+ dependent 1E (Ppm1e) were elevated in the parabrachial of diestrus animals both genes having a role in pain signaling. Epoxide hydroxylase (Ephx2) was elevated in the parabrachial at proestrus and the vitamin D receptor (Vdr) was elevated in the amygdala. Ephx2 antagonists and vitamin D have been used to treat neuropathic pain. In conclusion, sex steroids regulate genes in the parabrachial and amygdala that might result in the greater pain response observed during diestrus.<\/jats:p>","DOI":"10.3390\/ijms21165749","type":"journal-article","created":{"date-parts":[[2020,8,11]],"date-time":"2020-08-11T09:28:57Z","timestamp":1597138137000},"page":"5749","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Comparing Gene Expression in the Parabrachial and Amygdala of Diestrus and Proestrus Female Rats after Orofacial Varicella Zoster Injection"],"prefix":"10.3390","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7629-8144","authenticated-orcid":false,"given":"Rebecca","family":"Hornung","sequence":"first","affiliation":[{"name":"Department of Biological Sciences, Texas A&amp;M University College of Dentistry, Dallas, TX 75246, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Addison","family":"Pritchard","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Texas A&amp;M University College of Dentistry, Dallas, TX 75246, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1901-9970","authenticated-orcid":false,"given":"Paul R.","family":"Kinchington","sequence":"additional","affiliation":[{"name":"Departments of Ophthalmology and Molecular Microbiology and Genetics, University of Pittsburgh, 203 Lothrop Street, Pittsburgh, PA 15213, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0117-542X","authenticated-orcid":false,"given":"Phillip R.","family":"Kramer","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Texas A&amp;M University College of Dentistry, Dallas, TX 75246, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,8,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1016\/S0304-3959(00)00408-5","article-title":"Orofacial pain symptoms: An interaction between age and sex","volume":"90","author":"Riley","year":"2001","journal-title":"Pain"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1097\/00002508-200006000-00005","article-title":"Clinical pain perception and hormone replacement therapy in postmenopausal women experiencing orofacial pain","volume":"16","author":"Wise","year":"2000","journal-title":"Clin. J. Pain"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1016\/S0304-3959(96)03230-7","article-title":"Use of exogenous hormones and risk of temporomandibular disorder pain","volume":"69","author":"LeResche","year":"1997","journal-title":"Pain"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1016\/j.neuroscience.2014.07.066","article-title":"Infusion of Gabralpha6 siRNA into the trigeminal ganglia increased the myogenic orofacial nociceptive response of ovariectomized rats treated with 17beta-estradiol","volume":"278","author":"Kramer","year":"2014","journal-title":"Neuroscience"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"190","DOI":"10.1124\/jpet.110.167445","article-title":"17beta-estradiol rapidly enhances bradykinin signaling in primary sensory neurons in vitro and in vivo","volume":"335","author":"Rowan","year":"2010","journal-title":"J. Pharm. Exp."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1177\/0022034511428156","article-title":"Rapid Estrogenic Effects on TMJ-Responsive Brainstem Neurons","volume":"91","author":"Tashiro","year":"2011","journal-title":"J. Dent. Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1585","DOI":"10.1152\/jn.00269.2004","article-title":"Estrogen and inflammation increase the excitability of rat temporomandibular joint afferent neurons","volume":"93","author":"Flake","year":"2005","journal-title":"J. Neurophysiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"822","DOI":"10.1016\/j.neuroscience.2007.01.051","article-title":"Sex-related differences in NMDA-evoked rat masseter muscle afferent discharge result from estrogen-mediated modulation of peripheral NMDA receptor activity","volume":"146","author":"Dong","year":"2007","journal-title":"Neuroscience"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1002\/cne.903290205","article-title":"The organization of the efferent projections from the pontine parabrachial area to the amygdaloid complex: A Phaseolus vulgaris leucoagglutinin (PHA-L) study in the rat","volume":"329","author":"Bernard","year":"1993","journal-title":"J. Comp. Neurol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1016\/S0079-6123(08)61868-3","article-title":"Involvement of the spino-parabrachio -amygdaloid and -hypothalamic pathways in the autonomic and affective emotional aspects of pain","volume":"107","author":"Bernard","year":"1996","journal-title":"Prog. Brain Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1113\/eph8702357","article-title":"Pain pathways and parabrachial circuits in the rat","volume":"87","author":"Gauriau","year":"2002","journal-title":"Exp. Physiol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1016\/0006-8993(94)90025-6","article-title":"Progesterone metabolites, effective at the GABAA receptor complex, attenuate pain sensitivity in rats","volume":"643","author":"Frye","year":"1994","journal-title":"Brain Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"306","DOI":"10.1037\/0735-7044.118.2.306","article-title":"Estrogen and\/or progesterone administered systemically or to the amygdala can have anxiety-, fear-, and pain-reducing effects in ovariectomized rats","volume":"118","author":"Frye","year":"2004","journal-title":"Behav. Neurosci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1734","DOI":"10.1038\/s41593-017-0012-1","article-title":"A craniofacial-specific monosynaptic circuit enables heightened affective pain","volume":"20","author":"Rodriguez","year":"2017","journal-title":"Nat. Neurosci."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1016\/S0006-8993(03)02432-6","article-title":"Estrogen-induced autonomic effects are mediated by NMDA and GABAA receptors in the parabrachial nucleus","volume":"973","author":"Saleh","year":"2003","journal-title":"Brain Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"666","DOI":"10.1002\/jnr.20959","article-title":"17beta-estradiol attenuates excitatory neurotransmission and enhances the excitability of rat parabrachial neurons in vitro","volume":"84","author":"Fatehi","year":"2006","journal-title":"J. Neurosci. Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1002\/cne.20569","article-title":"Estrogen receptor-alpha and -beta immunoreactive neurons in the brainstem and spinal cord of male and female mice: Relationships to monoaminergic, cholinergic, and spinal projection systems","volume":"488","author":"Vanderhorst","year":"2005","journal-title":"J. Comp. Neurol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1016\/j.physbeh.2009.02.037","article-title":"Sex differences in the activation of the spinoparabrachial circuit by visceral pain","volume":"97","author":"Murphy","year":"2009","journal-title":"Physiol. Behav."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"627","DOI":"10.1056\/NEJM199009063231002","article-title":"Latent varicella-zoster viral DNA in human trigeminal and thoracic ganglia","volume":"323","author":"Mahalingam","year":"1990","journal-title":"N. Engl. J. Med."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"8151","DOI":"10.1128\/jvi.69.12.8151-8154.1995","article-title":"Configuration of latent varicella-zoster virus DNA","volume":"69","author":"Clarke","year":"1995","journal-title":"J. Virol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2122","DOI":"10.1073\/pnas.93.5.2122","article-title":"Expression of protein encoded by varicella-zoster virus open reading frame 63 in latently infected human ganglionic neurons","volume":"93","author":"Mahalingam","year":"1996","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"7080","DOI":"10.1073\/pnas.95.12.7080","article-title":"Aberrant intracellular localization of Varicella-Zoster virus regulatory proteins during latency","volume":"95","author":"Lungu","year":"1998","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Johnson, R.W., Bouhassira, D., Kassianos, G., Leplege, A., Schmader, K.E., and Weinke, T. (2010). The impact of herpes zoster and post-herpetic neuralgia on quality-of-life. BMC Med., 8.","DOI":"10.1186\/1741-7015-8-37"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"590","DOI":"10.1007\/s13365-011-0069-7","article-title":"Varicella zoster virus-induced pain and post-herpetic neuralgia in the human host and in rodent animal models","volume":"17","author":"Kinchington","year":"2011","journal-title":"J. Neurovirol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"959","DOI":"10.1212\/01.WNL.0000140708.62856.72","article-title":"Practice parameter: Treatment of postherpetic neuralgia: An evidence-based report of the Quality Standards Subcommittee of the American Academy of Neurology","volume":"63","author":"Dubinsky","year":"2004","journal-title":"Neurology"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1080\/13550280390194000","article-title":"Herpes simplex virus-1 and varicella-zoster virus latency in ganglia","volume":"9","author":"Mitchell","year":"2003","journal-title":"J. Neurovirol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2433","DOI":"10.1099\/0022-1317-80-9-2433","article-title":"Behavioural changes in the rat following infection with varicella-zoster virus","volume":"80","author":"Quinn","year":"1999","journal-title":"J. Gen. Virol."},{"key":"ref_28","first-page":"694","article-title":"Relief of pain induced by varicella-zoster virus in a rat model of post-herpetic neuralgia using a herpes simplex virus vector expressing enkephalin","volume":"21","author":"Guedon","year":"2014","journal-title":"Gene"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1093\/brain\/119.2.347","article-title":"The relationship of pain, allodynia and thermal sensation in post-herpetic neuralgia","volume":"119","author":"Rowbotham","year":"1996","journal-title":"Brain"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1016\/j.brainresrev.2004.07.008","article-title":"Allodynia in rats infected with varicella zoster virus\u2014A small animal model for post-herpetic neuralgia","volume":"46","author":"Dalziel","year":"2004","journal-title":"Brain Res. Rev."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Kennedy, P.G., Montague, P., Scott, F., Grinfeld, E., Ashrafi, G.H., Breuer, J., and Rowan, E.G. (2013). Varicella-zoster viruses associated with post-herpetic neuralgia induce sodium current density increases in the ND7-23 Nav-1.8 neuroblastoma cell line. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0051570"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Zhang, G.H., Lv, M.M., Wang, S., Chen, L., Qian, N.S., Tang, Y., Zhang, X.D., Ren, P.C., Gao, C.J., and Sun, X.D. (2011). Spinal astrocytic activation is involved in a virally-induced rat model of neuropathic pain. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0023059"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/j.pain.2005.08.003","article-title":"Varicella zoster virus induces neuropathic changes in rat dorsal root ganglia and behavioral reflex sensitisation that is attenuated by gabapentin or sodium channel blocking drugs","volume":"118","author":"Garry","year":"2005","journal-title":"Pain"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1495","DOI":"10.1016\/j.neuroscience.2006.11.029","article-title":"Further characterization of a rat model of varicella zoster virus-associated pain: Relationship between mechanical hypersensitivity and anxiety-related behavior, and the influence of analgesic drugs","volume":"144","author":"Hasnie","year":"2007","journal-title":"Neuroscience"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1016\/j.neuroscience.2019.06.029","article-title":"Estradiol Acts in Lateral Thalamic Region to Attenuate Varicella Zoster Virus Associated Affective Pain","volume":"414","author":"Stinson","year":"2019","journal-title":"Neuroscience"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1111\/j.1600-0714.2006.00489.x","article-title":"Evaluation of the sensation in patients with trigeminal post-herpetic neuralgia","volume":"36","author":"Alvarez","year":"2007","journal-title":"J. Oral Pathol. Med."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"178","DOI":"10.1016\/j.jinf.2014.08.018","article-title":"Incidence of herpes zoster and its complications in Germany, 2005\u20132009","volume":"70","author":"Hillebrand","year":"2014","journal-title":"J. Infect."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"502","DOI":"10.1038\/nrn3516","article-title":"Cognitive and emotional control of pain and its disruption in chronic pain","volume":"14","author":"Bushnell","year":"2013","journal-title":"Nat. Rev. Neurosci."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1704","DOI":"10.1210\/endo-94-6-1704","article-title":"Plasma concentration of LH, FSH, prolactin, progesterone and estradiol-17beta throughout the 4-day estrous cycle of the rat","volume":"94","author":"Butcher","year":"1974","journal-title":"Endocrinology"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Stinson, C., Deng, M., Yee, M.B., Bellinger, L.L., Kinchington, P.R., and Kramer, P.R. (2017). Sex differences underlying orofacial varicella zoster associated pain in rats. BMC Neurol., 17.","DOI":"10.1186\/s12883-017-0882-6"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"997","DOI":"10.1038\/nn.4037","article-title":"Distinct circuit-dependent functions of presynaptic neurexin-3 at GABAergic and glutamatergic synapses","volume":"18","author":"Aoto","year":"2015","journal-title":"Nat. Neurosci."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Oyang, E.L., Davidson, B.C., Lee, W., and Poon, M.M. (2011). Functional characterization of the dendritically localized mRNA neuronatin in hippocampal neurons. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0024879"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1186\/1423-0127-17-41","article-title":"Altered neuronatin expression in the rat dorsal root ganglion after sciatic nerve transection","volume":"17","author":"Chen","year":"2010","journal-title":"J. Biomed. Sci."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1016\/j.pharmthera.2017.06.006","article-title":"Soluble epoxide hydrolase as a therapeutic target for pain, inflammatory and neurodegenerative diseases","volume":"180","author":"Wagner","year":"2017","journal-title":"Pharmacol. Ther."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1016\/j.cellsig.2010.08.010","article-title":"Ppm1E is an in cellulo AMP-activated protein kinase phosphatase","volume":"23","author":"Voss","year":"2011","journal-title":"Cell. Signal."},{"key":"ref_46","first-page":"232","article-title":"Functional characterization of AMP-activated protein kinase signaling in tumorigenesis","volume":"1866","author":"Cheng","year":"2016","journal-title":"Biochim. Et Biophys. Acta"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"172909","DOI":"10.1016\/j.ejphar.2020.172909","article-title":"The intraperitoneal administration of MOTS-c produces antinociceptive and anti-inflammatory effects through the activation of AMPK pathway in the mouse formalin test","volume":"870","author":"Yin","year":"2020","journal-title":"Eur. J. Pharm."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1124\/jpet.119.258400","article-title":"Indirect AMP-Activated Protein Kinase Activators Prevent Incision-Induced Hyperalgesia and Block Hyperalgesic Priming, Whereas Positive Allosteric Modulators Block Only Priming in Mice","volume":"371","author":"Inyang","year":"2019","journal-title":"J. Pharm. Exp."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1016\/S0891-0618(99)00002-2","article-title":"Distribution of 1,25-dihydroxyvitamin D3 receptor immunoreactivity in the rat brain and spinal cord","volume":"16","author":"Veenstra","year":"1999","journal-title":"J. Chem. Neuroanat."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"7208","DOI":"10.1007\/s12035-019-1582-6","article-title":"Cholecalciferol (Vitamin D(3)) Reduces Rat Neuropathic Pain by Modulating Opioid Signaling","volume":"56","author":"Poisbeau","year":"2019","journal-title":"Mol. Neurobiol."},{"key":"ref_51","unstructured":"Paxinos, G., and Watson, C. (2007). The Rat Brain in Stereotaxic Coordinates, Elsevier Inc.. [6th ed.]."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1105","DOI":"10.1093\/bioinformatics\/btp120","article-title":"TopHat: Discovering splice junctions with RNA-Seq","volume":"25","author":"Trapnell","year":"2009","journal-title":"Bioinform. Oxf. Engl."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/16\/5749\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:59:04Z","timestamp":1760176744000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/16\/5749"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,8,11]]},"references-count":52,"journal-issue":{"issue":"16","published-online":{"date-parts":[[2020,8]]}},"alternative-id":["ijms21165749"],"URL":"https:\/\/doi.org\/10.3390\/ijms21165749","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,8,11]]}}}