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In rodents, RTN neurons can be histologically identified by the presence of paired-like homeobox 2B positive nuclei (Phox2b\u2009+) and the absence of cytoplasmic choline acetyltransferase (ChAT-) and tyrosine hydroxylase (TH-). Up to 50% of rodent RTN neurons synthesise galanin, and 88% express pituitary adenylate cyclase activating polypeptide (PACAP). The human RTN (hRTN) has not been mapped to date. This study aimed to map the location and cytoarchitecture of the adult hRTN and compare the findings to the homologies of rodents, macaques and human infants. Formalin-fixed, paraffin-embedded tissue blocks from two adult cases, spanning the medulla-pons, were serially sectioned (10\u00a0\u00b5m thick) and every four in thirty sections was assayed for immunohistochemistry for ChAT, or double-labelled Phox2b\/TH, Phox2b\/galanin and Phox2b\/PACAP, followed by analysis using QuPath software. hRTN neurons, identified as Phox2b\u2009+\u2009\/TH-\/ChAT-, were located ventral to 7N and lateral to the superior olive, overlapped with the C1 or A5 catecholaminergic population and extended rostrocaudally from Obex\u2009+\u200913 to\u2009+\u200917\u00a0mm. In the parafacial area, 90% of Phox2b immunoreactive (-ir) neurons are hRTN neurons, totaling around 5000 bilaterally, and were surrounded by numerous TH-ir fibers. Galanin- and PACAP-ir was identified in 43% and 39% of Phox2b-ir parafacial neurons, respectively. This is the first study to characterise and quantitatively map the adult human RTN using a series of neurochemical markers.<\/jats:p>","DOI":"10.1007\/s00429-025-02991-9","type":"journal-article","created":{"date-parts":[[2025,8,11]],"date-time":"2025-08-11T11:55:25Z","timestamp":1754913325000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Characterisation of putative retrotrapezoid nucleus (RTN) chemoreceptor neurons in the adult human brainstem"],"prefix":"10.1007","volume":"230","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2096-4064","authenticated-orcid":false,"given":"Yazhou","family":"Liu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7675-1121","authenticated-orcid":false,"given":"Rita","family":"Machaalani","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Irit","family":"Markus","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0399-3218","authenticated-orcid":false,"given":"Claire E.","family":"Shepherd","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4712-8792","authenticated-orcid":false,"given":"Natasha N.","family":"Kumar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,8,11]]},"reference":[{"key":"2991_CR1","doi-asserted-by":"publisher","first-page":"299","DOI":"10.1016\/j.resp.2009.06.003","volume":"167","author":"SB Abbott","year":"2009","unstructured":"Abbott SB, burke PG, Pilowsky PM (2009) Galanin microinjection into the PreB\u00f6tzinger or the B\u00f6tzinger Complex terminates central inspiratory activity and reduces responses to hypoxia and hypercapnia in rat. 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HC#200026. This research project (Project no. HC#200901) was approved by the Human Research Ethics Advisory Panel of the University of New South Wales.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethical approval"}},{"value":"Informed consent was obtained from individual participants included in the study prior to tissue donation.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent to participate"}},{"value":"The authors affirm that human research participants provided informed consent for use of their data for publication.","order":5,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent to publish"}}],"article-number":"131"}}