{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T18:57:46Z","timestamp":1785437866348,"version":"3.56.0"},"posted":{"date-parts":[[2025,5,1]]},"group-title":"Pharmacology and Toxicology","reference-count":95,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,5,1]],"date-time":"2025-05-01T00:00:00Z","timestamp":1746057600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/publicdomain\/zero\/1.0\/"}],"funder":[{"award":["DA-000522-16"],"award-info":[{"award-number":["DA-000522-16"]}],"id":[{"id":"https:\/\/ror.org\/00fq5cm18","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,7,16]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Serotonergic psychedelics induce the head twitch response (HTR) in mice, an index of serotonin (5-HT) 2A receptor (5-HT\n                  <jats:sub>2A<\/jats:sub>\n                  ) agonism and a behavioral proxy for psychedelic effects in humans. Existing methods for detecting HTRs include time-consuming visual scoring, magnetometer-based approaches, and analysis of videos using semi-automated commercial software. Here, we present a new automated approach for quantifying HTRs from experimental videos using the open-source machine learning-based toolkits, DeepLabCut (DLC) and Simple Behavioral Analysis (SimBA). Pose estimation DLC models were trained to predict X,Y coordinates of 13 body parts of C57BL\/6J mice using historical experimental videos of HTRs induced by various psychedelic drugs. Next, a non-overlapping set of historical experimental videos was analyzed and used to train SimBA random forest behavioral classifiers to predict the presence of the HTR. The DLC+SimBA approach was then validated using a separate subset of visually scored videos. DLC+SimBA model performance was assessed at different video resolutions (50%, 25%, 12.5%) and frame rates (120, 60, 30 frames per second or fps). Our results indicate that HTRs can be quantified accurately at 50% resolution and 120 fps (precision = 95.45, recall = 95.56, F\n                  <jats:sub>1<\/jats:sub>\n                  = 95.51) or at lower frame rates and resolutions (i.e., 50% resolution and 60 fps). The best performing DLC+SimBA model combination was deployed to evaluate the effects of bufotenine, a tryptamine derivative with uncharacterized potency and efficacy in the HTR paradigm. Interestingly, bufotenine only induced elevated HTRs (ED\n                  <jats:sub>50<\/jats:sub>\n                  = 0.99 mg\/kg, max counts = 24) when serotonin 1A receptors (5-HT\n                  <jats:sub>1A<\/jats:sub>\n                  ) were pharmacologically blocked and activity at other sites of action may also impact its pharmacological effects (e.g., serotonin transporter). HTR counts for a subset of 21 videos from bufotenine experiments were strongly correlated for DLC+SimBA vs. visual scoring and semi-automated software detection methods (\n                  <jats:italic>r<\/jats:italic>\n                  = 0.98 and 0.99). Finally, the DLC+SimBA approach displayed high accuracy when compared to visual scoring of HTRs for three serotonergic psychedelic drugs with variable HTR frequencies (\n                  <jats:italic>r<\/jats:italic>\n                  = 0.99 vs. mean visual scores from 3 blinded raters). In summary, the DLC+SimBA approach represents a modular, noninvasive, and open-source method of HTR detection from experimental videos with accuracy comparable to magnetometer-based approaches and greater speed than visual scoring.\n                <\/jats:p>","DOI":"10.1101\/2025.04.28.650242","type":"posted-content","created":{"date-parts":[[2025,5,1]],"date-time":"2025-05-01T15:05:11Z","timestamp":1746111911000},"source":"Crossref","is-referenced-by-count":1,"title":["Rapid, open-source, and automated quantification of the head twitch response in C57BL\/6J mice using DeepLabCut and Simple Behavioral Analysis"],"prefix":"10.64898","author":[{"given":"Alexander D.","family":"Maitland","sequence":"first","affiliation":[{"name":"Designer Drug Research Unit, National Institute on Drug Abuse, Intramural Research Program, Baltimore, Maryland 21224, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicholas R.","family":"Gonzalez","sequence":"additional","affiliation":[{"name":"Designer Drug Research Unit, National Institute on Drug Abuse, Intramural Research Program, Baltimore, Maryland 21224, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Donna","family":"Walther","sequence":"additional","affiliation":[{"name":"Designer Drug Research Unit, National Institute on Drug Abuse, Intramural Research Program, Baltimore, Maryland 21224, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francisco","family":"Pereira","sequence":"additional","affiliation":[{"name":"Machine Learning Team, National Institute of Mental Health, Intramural Research Program, Bethesda, Maryland 20814, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael H.","family":"Baumann","sequence":"additional","affiliation":[{"name":"Designer Drug Research Unit, National Institute on Drug Abuse, Intramural Research Program, Baltimore, Maryland 21224, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Grant C.","family":"Glatfelter","sequence":"additional","affiliation":[{"name":"Designer Drug Research Unit, National Institute on Drug Abuse, Intramural Research Program, Baltimore, Maryland 21224, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026041206260771000_2025.04.28.650242v2.1","doi-asserted-by":"publisher","DOI":"10.1007\/s00213-013-3006-z"},{"key":"2026041206260771000_2025.04.28.650242v2.2","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuron.2007.01.008"},{"key":"2026041206260771000_2025.04.28.650242v2.3","doi-asserted-by":"publisher","DOI":"10.1002\/dta.1333"},{"key":"2026041206260771000_2025.04.28.650242v2.4","doi-asserted-by":"publisher","DOI":"10.1124\/pr.115.011478"},{"key":"2026041206260771000_2025.04.28.650242v2.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuropharm.2019.107933"},{"key":"2026041206260771000_2025.04.28.650242v2.6","doi-asserted-by":"publisher","DOI":"10.1021\/acschemneuro.4c00513"},{"key":"2026041206260771000_2025.04.28.650242v2.7","doi-asserted-by":"publisher","DOI":"10.1021\/acsptsci.2c00177"},{"key":"2026041206260771000_2025.04.28.650242v2.8","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jmedchem.3c01961"},{"key":"2026041206260771000_2025.04.28.650242v2.9","doi-asserted-by":"publisher","DOI":"10.1021\/cn500292d"},{"key":"2026041206260771000_2025.04.28.650242v2.10","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuropharm.2013.08.025"},{"key":"2026041206260771000_2025.04.28.650242v2.11","doi-asserted-by":"publisher","DOI":"10.1021\/acschemneuro.3c00610"},{"key":"2026041206260771000_2025.04.28.650242v2.12","doi-asserted-by":"publisher","DOI":"10.1124\/jpet.110.172247"},{"key":"2026041206260771000_2025.04.28.650242v2.13","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-020-3008-z"},{"key":"2026041206260771000_2025.04.28.650242v2.14","doi-asserted-by":"publisher","DOI":"10.1016\/S0031-9384(96)00323-X"},{"key":"2026041206260771000_2025.04.28.650242v2.15","doi-asserted-by":"publisher","DOI":"10.1016\/j.biopha.2022.113612"},{"key":"2026041206260771000_2025.04.28.650242v2.16","doi-asserted-by":"publisher","DOI":"10.1007\/s00213-010-1784-0"},{"key":"2026041206260771000_2025.04.28.650242v2.17","doi-asserted-by":"publisher","DOI":"10.3791\/53717"},{"key":"2026041206260771000_2025.04.28.650242v2.18","first-page":"3","article-title":"Department of Health and Human Services: Office of Disease Prevention and Health Promotion--Healthy People","volume":"15","author":"U.S","year":"2020","journal-title":"Nasnewsletter"},{"key":"2026041206260771000_2025.04.28.650242v2.19","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-019-49913-4"},{"key":"2026041206260771000_2025.04.28.650242v2.20","doi-asserted-by":"crossref","unstructured":"Jaster, A. 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