{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,20]],"date-time":"2026-02-20T23:49:19Z","timestamp":1771631359707,"version":"3.50.1"},"reference-count":37,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2023,6,23]],"date-time":"2023-06-23T00:00:00Z","timestamp":1687478400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000055","name":"National Institute on Deafness and Other Communication Disorders","doi-asserted-by":"publisher","award":["1R01DC018060"],"award-info":[{"award-number":["1R01DC018060"]}],"id":[{"id":"10.13039\/100000055","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Laryngoscope"],"published-print":{"date-parts":[[2023,12]]},"abstract":"<jats:sec><jats:title>Objectives<\/jats:title><jats:p>Recurrent laryngeal nerve (RLN) injury results in synkinetic reinnervation and vocal fold paralysis. Investigation of cues expressed in the developing brainstem that influence correct selective targeting of intrinsic laryngeal muscles may elucidate post\u2010injury abnormalities contributing to non\u2010functional reinnervation. Primary targets of interest were <jats:italic>Hoxb1<\/jats:italic> and <jats:italic>Hoxb2<\/jats:italic>, members of the Hox family that create overlapping gradients in the developing brain, and their target Phox2b, a transcription factor necessary for cranial nerve branchio\u2010 and visceromotoneuron survival.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Rat embryos at developmental days E14, E16, E18, and E20 (4 animals\/age) were sectioned for RNA <jats:italic>in situ<\/jats:italic> hybridization to detect <jats:italic>Hoxb1<\/jats:italic>, <jats:italic>Hoxb2<\/jats:italic>, and <jats:italic>Phox2b<\/jats:italic> mRNA within the brainstem. Slides were costained with Islet1 antibody for identification of the nucleus ambiguus. Results were confirmed using immunohistochemistry. Sections were imaged on a confocal microscope. RNA and protein expressions were quantified using QuPath. Statistical analyses were performed using R.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p><jats:italic>Hoxb1<\/jats:italic>, <jats:italic>Hoxb2<\/jats:italic>, and <jats:italic>Phox2b<\/jats:italic> expressions varied according to embryologic age. <jats:italic>Hoxb1<\/jats:italic> and <jats:italic>Hoxb2<\/jats:italic> expression peaked at E16, with significant decreases at E18 and E20 (one\u2010way ANOVA <jats:italic>p<\/jats:italic>\u2009=\u20090.001 for both). <jats:italic>Phox2b<\/jats:italic> expression was highest at E14 and trended downward with increased embryologic age (one\u2010way ANOVA <jats:italic>p<\/jats:italic>\u2009=\u20090.005).<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Peak expression of <jats:italic>Hoxb1<\/jats:italic> and <jats:italic>Hoxb2<\/jats:italic> is observed at time points when the RLN arrives at the larynx and begins to branch toward individual muscles, positioning these gene products to be involved in cueing laryngeal motoneuron identity and target identification. Higher expression of <jats:italic>Phox2b<\/jats:italic> earlier in development suggests a role in laryngeal motoneuron formation.<\/jats:p><\/jats:sec><jats:sec><jats:title>Level of Evidence<\/jats:title><jats:p>NA <jats:italic>Laryngoscope<\/jats:italic>, 133:3462\u20133471, 2023<\/jats:p><\/jats:sec>","DOI":"10.1002\/lary.30826","type":"journal-article","created":{"date-parts":[[2023,6,25]],"date-time":"2023-06-25T00:22:54Z","timestamp":1687652574000},"page":"3462-3471","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Temporal Expression of Hox Genes and Phox2b in the Rat Nucleus Ambiguus During Development: Implications on Laryngeal Innervation"],"prefix":"10.1002","volume":"133","author":[{"given":"Emily","family":"Honzel","sequence":"first","affiliation":[{"name":"Department of Otolaryngology\u2010Head &amp; Neck Surgery Columbia University College of Physicians and Surgeons  New York New York U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3628-030X","authenticated-orcid":false,"given":"Ignacio","family":"Hernandez\u2010Morato","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology\u2010Head &amp; Neck Surgery Columbia University College of Physicians and Surgeons  New York New York U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Abhinav","family":"Joshi","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology\u2010Head &amp; Neck Surgery Columbia University College of Physicians and Surgeons  New York New York U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"William","family":"Pennington\u2010Fitzgerald","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology\u2010Head &amp; Neck Surgery Columbia University College of Physicians and Surgeons  New York New York U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0600-1253","authenticated-orcid":false,"given":"Yalda","family":"Moayedi","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology\u2010Head &amp; Neck Surgery Columbia University College of Physicians and Surgeons  New York New York U.S.A."},{"name":"Department of Neurology Columbia University  New York New York U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6637-4302","authenticated-orcid":false,"given":"Michael J.","family":"Pitman","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology\u2010Head &amp; Neck Surgery Columbia University College of Physicians and Surgeons  New York New York U.S.A."},{"name":"Department of Neurology Columbia University  New York New York U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2023,6,23]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/ans.12980"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/hed.24772"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0030-6665(03)00172-5"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1097\/MD.0000000000006674"},{"key":"e_1_2_8_6_1","first-page":"27","article-title":"Recovery of laryngeal function after intraoperative injury to the recurrent laryngeal nerve","volume":"4","author":"Mattsson P","year":"2015","journal-title":"Gland Surg"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1002\/lary.24759"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/lary.24216"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuroscience.2016.07.004"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1002\/lary.30440"},{"key":"e_1_2_8_11_1","unstructured":"SasakiCT.Anatomy and development and physiology of the larynx. 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