{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T18:15:31Z","timestamp":1785435331219,"version":"3.56.0"},"posted":{"date-parts":[[2026,2,16]]},"group-title":"Neuroscience","reference-count":32,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,2,16]],"date-time":"2026-02-16T00:00:00Z","timestamp":1771200000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"award":["R01NS125413"],"award-info":[{"award-number":["R01NS125413"]}],"id":[{"id":"https:\/\/ror.org\/01s5ya894","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,2,16]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Chronic itch is a major burden, impacting the quality of life for one in four adults, and is closely associated with increased levels of anxiety, depression, and suicide. Despite its widespread burden, the precise mechanisms driving and sustaining chronic itch remain poorly understood. Non-histaminergic itch transmission in the spinal cord relies on the synaptic vesicle (SV) release of gastrin-releasing peptide (GRP) and the subsequent binding to the Gastrin-Releasing Peptide Receptor (GRPR). While SV exocytosis facilitates neurotransmitter release, such as GRP, SV endocytosis, mediated by key proteins including clathrin, adaptor associated kinase 1 (AAK1), and Dynamin (Dnm) are essential for retrieving and recycling SV from the presynaptic membrane to maintain signaling. Building upon evidence that AAK1- and Dnm-mediated endocytosis are viable targets to reverse pain, we characterized the role of these endocytic mediators in the regulation of synaptic transmission in itch pathways. We localized mRNA encoding AAK1, Dnm1 and Dnm3 within GRP positive neurons of the mouse dorsal root ganglia (DRG). Genetic and pharmacological disruption of AAK1 significantly reduced scratching behavior compared to control groups. This anti-pruritic effect correlated with confirmed\n                  <jats:italic>Aak1<\/jats:italic>\n                  mRNA knockdown in both the DRG and spinal cord. Similarly, siRNA mediated knockdown of Dnm1 and Dnm3 in the DRGs also reduced scratching behavior. Crucially, these treatments decreased GRP release without altering locomotor activity or anxiety-like behaviors. Together, these findings suggest that the disruption of SV recycling reduces itch related signaling and behavior without affecting normal motor functions, providing a new approach for chronic pruritus.\n                <\/jats:p>","DOI":"10.64898\/2026.02.13.705766","type":"posted-content","created":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T05:25:17Z","timestamp":1771305917000},"source":"Crossref","is-referenced-by-count":0,"title":["The contribution of endocytic mediators to itch transmission"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3804-9662","authenticated-orcid":false,"given":"Marcella","family":"de A. Ferreira","sequence":"first","affiliation":[{"name":"Pain Research Center, College of Dentistry New York University, New York, NY"},{"name":"Translational Research Center, College of Dentistry, New York University, New York, NY, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2175-9804","authenticated-orcid":false,"given":"Raquel","family":"Tonello","sequence":"additional","affiliation":[{"name":"Pain Research Center, College of Dentistry New York University, New York, NY"},{"name":"Department of Molecular Pathobiology, College of Dentistry, New York University, New York, NY, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8729-8636","authenticated-orcid":false,"given":"Paz","family":"Duran","sequence":"additional","affiliation":[{"name":"Pain Research Center, College of Dentistry New York University, New York, NY"},{"name":"Translational Research Center, College of Dentistry, New York University, New York, NY, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kai","family":"Trevett","sequence":"additional","affiliation":[{"name":"Pain Research Center, College of Dentistry New York University, New York, NY"},{"name":"Department of Molecular Pathobiology, College of Dentistry, New York University, New York, NY, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6540-2375","authenticated-orcid":false,"given":"Dane D.","family":"Jensen","sequence":"additional","affiliation":[{"name":"Pain Research Center, College of Dentistry New York University, New York, NY"},{"name":"Translational Research Center, College of Dentistry, New York University, New York, NY, United States"},{"name":"Department of Molecular Pathobiology, College of Dentistry, New York University, New York, NY, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026021901100943000_2026.02.13.705766v1.1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.2559-18.2019"},{"key":"2026021901100943000_2026.02.13.705766v1.2","doi-asserted-by":"crossref","unstructured":"Barry DM , Li H , Liu XY , Shen KF , Liu XT , Wu ZY , Munanairi A , Chen XJ , Yin J , Sun YG , Li YQ , Chen ZF . Critical evaluation of the expression of gastrin-releasing peptide in dorsal root ganglia and spinal cord. Mol Pain 2016;12.","DOI":"10.1177\/1744806916643724"},{"issue":"1","key":"2026021901100943000_2026.02.13.705766v1.3","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1007\/s12264-017-0125-2","article-title":"Spinal Mechanisms of Itch Transmission","volume":"34","year":"2018","journal-title":"Neuroscience Bulletin"},{"key":"2026021901100943000_2026.02.13.705766v1.4","doi-asserted-by":"publisher","DOI":"10.1016\/j.jneumeth.2013.12.010"},{"issue":"6","key":"2026021901100943000_2026.02.13.705766v1.5","doi-asserted-by":"crossref","first-page":"1029","DOI":"10.1016\/j.clinthera.2021.04.014","article-title":"Results of two Phase 1, Randomized, Double-blind, Placebocontrolled, Studies (Ascending Single-dose and Multiple-dose Studies) to Determine the Safety, Tolerability, and Pharmacokinetics of Orally Administered LX9211 in Healthy Participants","volume":"43","year":"2021","journal-title":"Clin Ther"},{"key":"2026021901100943000_2026.02.13.705766v1.6","doi-asserted-by":"publisher","DOI":"10.1097\/j.pain.0000000000000280"},{"key":"2026021901100943000_2026.02.13.705766v1.7","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2020.02.019"},{"key":"2026021901100943000_2026.02.13.705766v1.8","doi-asserted-by":"publisher","DOI":"10.1016\/j.neulet.2021.136288"},{"key":"2026021901100943000_2026.02.13.705766v1.9","doi-asserted-by":"publisher","DOI":"10.1111\/tra.12272"},{"issue":"1","key":"2026021901100943000_2026.02.13.705766v1.10","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1186\/s12974-020-01807-3","article-title":"Spinal cord NLRP1 inflammasome contributes to dry skin induced chronic itch in mice","volume":"17","year":"2020","journal-title":"J Neuroinflammation"},{"key":"2026021901100943000_2026.02.13.705766v1.11","doi-asserted-by":"publisher","DOI":"10.1038\/nrm3266"},{"key":"2026021901100943000_2026.02.13.705766v1.12","doi-asserted-by":"crossref","unstructured":"Go EJ , Lee JY , Kim YH , Park CK . Site-Specific Transient Receptor Potential Channel Mechanisms and Their Characteristics for Targeted Chronic Itch Treatment. Biomolecules 2024;14(1).","DOI":"10.3390\/biom14010107"},{"key":"2026021901100943000_2026.02.13.705766v1.13","doi-asserted-by":"crossref","unstructured":"Hegron A , Peach CJ , Tonello R , Seemann P , Teng S , Latorre R , Huebner H , Weikert D , Rientjes J , Veldhuis NA , Poole DP , Jensen DD , Thomsen ARB , Schmidt BL , Imlach WL , Gmeiner P , Bunnett NW . Therapeutic antagonism of the neurokinin 1 receptor in endosomes provides sustained pain relief. Proceedings of the National Academy of Sciences of the United States of America 2023;120(22).","DOI":"10.1073\/pnas.2220979120"},{"issue":"10","key":"2026021901100943000_2026.02.13.705766v1.14","doi-asserted-by":"crossref","first-page":"750","DOI":"10.1111\/exd.13078","article-title":"Preclinical and human surrogate models of itch","volume":"25","author":"H HA","year":"2016","journal-title":"Exp Dermatol"},{"key":"2026021901100943000_2026.02.13.705766v1.15","doi-asserted-by":"crossref","unstructured":"Jensen DD , Lieu T , Halls ML , Veldhuis NA , Imlach WL , Mai QN , Poole DP , Quach T , Aurelio L , Conner J , Herenbrink CK , Barlow N , Simpson JS , Scanlon MJ , Graham B , McCluskey A , Robinson PJ , Escriou V , Nassini R , Materazzi S , Geppetti P , Hicks GA , Christie MJ , Porter CJH , Canals M , Bunnett NW . Neurokinin 1 receptor signaling in endosomes mediates sustained nociception and is a viable therapeutic target for prolonged pain relief. 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Characterization and targeting of the endosomal signaling of the gastrin releasing peptide receptor in pruritus. bioRxiv 2025.","DOI":"10.1101\/2025.03.17.643743"},{"key":"2026021901100943000_2026.02.13.705766v1.27","first-page":"1873","article-title":"The vicious cycle of itch and anxiety","volume":"87","year":"2018","journal-title":"Neuroscience and Biobehavioral Reviews"},{"key":"2026021901100943000_2026.02.13.705766v1.28","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.2955-14.2015"},{"key":"2026021901100943000_2026.02.13.705766v1.29","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0079123"},{"key":"2026021901100943000_2026.02.13.705766v1.30","doi-asserted-by":"publisher","DOI":"10.1038\/nature06029"},{"key":"2026021901100943000_2026.02.13.705766v1.31","doi-asserted-by":"publisher","DOI":"10.1097\/j.pain.0000000000002826"},{"key":"2026021901100943000_2026.02.13.705766v1.32","doi-asserted-by":"crossref","unstructured":"Xu K , Liu X , Zeng Q , Liu Y , Shan L , Ji L , Wu Y , Wu J , Chen Y , Li Y , Huang S , Jiang C , Hong X , Wu C , Wang Z. 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