{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T19:42:06Z","timestamp":1785267726912,"version":"3.55.0"},"reference-count":34,"publisher":"Public Library of Science (PLoS)","issue":"4","license":[{"start":{"date-parts":[[2024,4,16]],"date-time":"2024-04-16T00:00:00Z","timestamp":1713225600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000038","name":"Natural Sciences and Engineering Research Council of Canada","doi-asserted-by":"publisher","award":["458021"],"award-info":[{"award-number":["458021"]}],"id":[{"id":"10.13039\/501100000038","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000038","name":"Natural Sciences and Engineering Research Council of Canada","doi-asserted-by":"publisher","award":["478124"],"award-info":[{"award-number":["478124"]}],"id":[{"id":"10.13039\/501100000038","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Anoxia in the mammalian brain leads to hyper-excitability and cell death; however, this cascade of events does not occur in the anoxia-tolerant brain of the western painted turtle, <jats:italic>Chrysemys picta belli<\/jats:italic>. The painted turtle has become an important anoxia-tolerant model to study brain, heart, and liver function in the absence of oxygen, but being anoxia-tolerant likely means that decapitation alone is not a suitable method of euthanasia. Many anesthetics have long-term effects on ion channels and are not appropriate for same day experimentation. Using whole-cell electrophysiological techniques, we examine the effects of the anesthetic, Alfaxalone, on pyramidal cell action potential amplitude, threshold, rise and decay time, width, frequency, whole cell conductance, and evoked GABA<jats:sub>A<\/jats:sub> receptors currents to determine if any of these characteristics are altered with the use of Alfaxalone for animal sedation. We find that Alfaxalone has no long-term impact on action potential parameters or whole-cell conductance. When acutely applied to na\u00efve tissue, Alfaxalone did lengthen GABA<jats:sub>A<\/jats:sub> receptor current decay rates by 1.5-fold. Following whole-animal sedation with Alfaxalone, evoked whole cell GABA<jats:sub>A<\/jats:sub> receptor current decay rates displayed an increasing trend with 1 and 2 hours after brain sheet preparation, but showed no significant change after a 3-hour washout period. Therefore, we conclude that Alfaxalone is a suitable anesthetic for same day use in electrophysiological studies in western painted turtle brain tissue.<\/jats:p>","DOI":"10.1371\/journal.pone.0298065","type":"journal-article","created":{"date-parts":[[2024,4,16]],"date-time":"2024-04-16T18:23:41Z","timestamp":1713291821000},"page":"e0298065","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":4,"title":["Alfaxalone is an effective anesthetic for the electrophysiological study of anoxia-tolerance mechanisms in western painted turtle pyramidal neurons"],"prefix":"10.1371","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-4586-3055","authenticated-orcid":true,"given":"Haushe","family":"Suganthan","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Domenic Di","family":"Stefano","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leslie T.","family":"Buck","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2024,4,16]]},"reference":[{"issue":"1","key":"pone.0298065.ref001","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1146\/annurev.physiol.69.031905.162529","article-title":"Hypoxia Tolerance in Reptiles, Amphibians, and Fishes: Life with Variable Oxygen Availability","volume":"69","author":"P. 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Buck","year":"2018","journal-title":"Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology"},{"issue":"4","key":"pone.0298065.ref005","first-page":"256","article-title":"Considerations for the Use of Anesthetics in Neurotoxicity Studies","volume":"60","author":"S. W. Karmarkar","year":"2010","journal-title":"Comparative Medicine"},{"issue":"7","key":"pone.0298065.ref006","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0101606","article-title":"Efficacy of Tricaine Methanesulfonate (MS-222) as an Anesthetic Agent for Blocking Sensory-Motor Responses in Xenopus laevis Tadpoles","volume":"9","author":"C. Ramlochansingh","year":"2014","journal-title":"PLoS ONE"},{"issue":"23","key":"pone.0298065.ref007","first-page":"4370","article-title":"Cold block ofin vitroeyeblink reflexes: evidence supporting the use of hypothermia as an anesthetic in pond turtles","volume":"220","author":"J. Keifer","year":"2017","journal-title":"The Journal of Experimental Biology"},{"issue":"3","key":"pone.0298065.ref008","first-page":"282","article-title":"A Review of Anesthesia and Chemical Restraint in Reptiles","volume":"22","author":"A. Bennett","year":"1991","journal-title":"Journal of Zoo and Wildlife Medicine"},{"issue":"6","key":"pone.0298065.ref009","doi-asserted-by":"crossref","first-page":"370","DOI":"10.1111\/cns.12099","article-title":"Ketamine Pharmacology: An Update (Pharmacodynamics and Molecular Aspects, Recent Findings)","volume":"19","author":"G. Mion","year":"2013","journal-title":"CNS Neuroscience & Therapeutics"},{"issue":"12","key":"pone.0298065.ref010","doi-asserted-by":"crossref","first-page":"1539","DOI":"10.1007\/s40262-018-0672-3","article-title":"Clinical Pharmacokinetics and Pharmacodynamics of Propofol","volume":"57","author":"M. M. Sahinovic","year":"2018","journal-title":"Clinical Pharmacokinetics"},{"key":"pone.0298065.ref011","author":"R. K. Stoelting","year":"1989","journal-title":"Basics of Anesthesia"},{"issue":"2","key":"pone.0298065.ref012","doi-asserted-by":"crossref","first-page":"699","DOI":"10.1093\/icb\/28.2.699","article-title":"Structural and functional determinates of metabolic rate","volume":"28","author":"A. Bennett","year":"1988","journal-title":"American Zoologist"},{"issue":"5","key":"pone.0298065.ref013","first-page":"R350","article-title":"Comparison of the \"mammal machine\" and the \"reptile machine\": energy use and thyroid activity","volume":"241","author":"AJ Hulbert","year":"1981","journal-title":"Am J Physiol"},{"key":"pone.0298065.ref014","unstructured":"Jurox. (2018, June 6). Freedom of Information Summary."},{"issue":"1","key":"pone.0298065.ref015","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1093\/bja\/43.1.2-a","article-title":"The Pharmacological Properties In Animals Of Ct1341\u2014A New Steroid Anesthetic Agent","volume":"43","author":"K. Child","year":"1971","journal-title":"British Journal of Anaesthesia"},{"issue":"7","key":"pone.0298065.ref016","doi-asserted-by":"crossref","first-page":"543","DOI":"10.1016\/j.tips.2016.04.003","article-title":"Clinical Potential of Neurosteroids for CNS Disorders","volume":"37","author":"D. S. Reddy","year":"2016","journal-title":"Trends in Pharmacological Sciences"},{"issue":"2","key":"pone.0298065.ref017","doi-asserted-by":"crossref","first-page":"604","DOI":"10.1523\/JNEUROSCI.07-02-00604.1987","article-title":"A steroid anesthetic prolongs inhibitory postsynaptic currents in cultured rat hippocampal neurons","volume":"7","author":"N. Harrison","year":"1987","journal-title":"The Journal of Neuroscience"},{"issue":"1","key":"pone.0298065.ref018","article-title":"Enhanced gabaergic actions resulting from the coapplication of the steroid 3\u03b1-hydroxy-5\u03b1-pregnane-11,20-dione (Alfaxalone) with propofol or diazepam","volume":"8","author":"L. Q. Cao","year":"2018","journal-title":"Scientific Reports"},{"issue":"1","key":"pone.0298065.ref019","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1111\/j.1467-2995.2009.00503.x","article-title":"Alfaxalone induction dose following administration of Medetomidine and butorphanol in the dog","volume":"37","author":"K. Maddern","year":"2010","journal-title":"Veterinary Anaesthesia and Analgesia"},{"issue":"6","key":"pone.0298065.ref020","doi-asserted-by":"crossref","first-page":"590","DOI":"10.1111\/vaa.12061","article-title":"Pharmacodynamics of Alfaxalone after single\u2010dose intramuscular administration in red\u2010eared sliders (Trachemys scripta elegans): a comparison of two different doses at two different ambient temperatures","volume":"40","author":"M. K. Shepard","year":"2013","journal-title":"Veterinary Anaesthesia and Analgesia"},{"key":"pone.0298065.ref021","unstructured":"Knotek, Z. (2015). Optimal Protocol for Intravenous Introduction to Inhalant Anaesthesia in Chelonians. World Small Animal Veterinary Association World Congress Proceedings."},{"issue":"3","key":"pone.0298065.ref022","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1016\/j.vaa.2017.12.003","article-title":"Anesthetic induction with alfaxalone in the ball Python (python regius): Dose response and effect of injection site","volume":"45","author":"L. E. James","year":"2018","journal-title":"Veterinary Anaesthesia and Analgesia"},{"key":"pone.0298065.ref023","doi-asserted-by":"crossref","first-page":"109","DOI":"10.2754\/avb201382010109","article-title":"Alfaxalone anaesthesia in the green iguana (Iguana iguana)","volume":"82","author":"Z Knotek","year":"2013","journal-title":"Acta Veterinaria Brno"},{"issue":"2","key":"pone.0298065.ref024","doi-asserted-by":"crossref","first-page":"466","DOI":"10.1152\/jn.00104.2019","article-title":"Oxygen-sensitive interneurons exhibit increased activity and GABA release during ROS scavenging in the cerebral cortex of the western painted turtle","volume":"122","author":"P. J. Hawrysh","year":"2019","journal-title":"Journal of Neurophysiology"},{"key":"pone.0298065.ref025","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1016\/0165-0270(89)90131-3","article-title":"Whole cell recording from neurons in slices of reptilian and mammalian cerebral cortex","volume":"30","author":"M. G. Blanton","year":"1989","journal-title":"J. Neurosci. Meth"},{"issue":"27","key":"pone.0298065.ref026","doi-asserted-by":"crossref","first-page":"11274","DOI":"10.1073\/pnas.1102429108","article-title":"Endogenous GABAA and GABAB receptor-mediated electrical suppression is critical to neuronal anoxia tolerance","volume":"108","author":"M. E. Pamenter","year":"2011","journal-title":"Proceedings of the National Academy of Sciences"},{"issue":"1","key":"pone.0298065.ref027","doi-asserted-by":"crossref","first-page":"164","DOI":"10.1523\/JNEUROSCI.06-01-00164.1986","article-title":"Cellular physiology of the turtle visual cortex: distinctive properties of pyramidal and stellate neurons","volume":"6","author":"B. Connors","year":"1986","journal-title":"The Journal of Neuroscience"},{"issue":"2","key":"pone.0298065.ref028","doi-asserted-by":"crossref","first-page":"1254","DOI":"10.1152\/jn.00346.2009","article-title":"The Influence of Neuroactive Steroid Lipophilicity on GABAA Receptor Modulation: Evidence for a Low-Affinity Interaction","volume":"102","author":"M. Chisari","year":"2009","journal-title":"Journal of Neurophysiology"},{"issue":"2","key":"pone.0298065.ref029","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1007\/s12035-007-8001-0","article-title":"Ionic Channel Function in Action Potential Generation: Current Perspective","volume":"35","author":"G. Baranauskas","year":"2007","journal-title":"Mol Neurobiol"},{"issue":"1","key":"pone.0298065.ref030","article-title":"Enhanced gabaergic actions resulting from the coapplication of the steroid 3\u03b1-hydroxy-5\u03b1-pregnane-11,20-dione (Alfaxalone) with propofol or diazepam","volume":"8","author":"M Chen","year":"2018","journal-title":"Scientific Reports"},{"issue":"3","key":"pone.0298065.ref031","first-page":"280","article-title":"Structurally Specific Effects of Some Steroid Anesthetics on Spin-Labeled Liposomes","volume":"11","author":"DK Lawrence","year":"1975","journal-title":"Mol Pharmacol"},{"key":"pone.0298065.ref032","first-page":"P83","article-title":"Voltage-Clamp Studies of the Potentiation of Gaba-Activated Chloride Conductance by the Steroid Anesthetic Alphaxalone and a Reduced Metabolite of Progesterone in Cultured Rat Cns Neurons","volume":"377","author":"JL Barker","year":"1986","journal-title":"Journal of Physiology-London"},{"issue":"6","key":"pone.0298065.ref033","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1016\/0165-6147(87)90067-8","article-title":"Actions of synthetic and endogenous steroids on the GABAA receptor","volume":"8","author":"JJ Lambert","year":"1987","journal-title":"Trends in Pharmacological Sciences"},{"issue":"5","key":"pone.0298065.ref034","doi-asserted-by":"crossref","first-page":"516","DOI":"10.1111\/jvp.12055","article-title":"Plasma pharmacokinetics of alfaxalone after a single intraperitoneal or intravenous injection of Alfaxan\u00ae in rats","volume":"36","author":"C Lau","year":"2013","journal-title":"Journal of Veterinary Pharmacology and Therapeutics"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0298065","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,4,16]],"date-time":"2024-04-16T18:24:41Z","timestamp":1713291881000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0298065"}},"subtitle":[],"editor":[{"given":"Steven","family":"Barnes","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2024,4,16]]},"references-count":34,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2024,4,16]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0298065","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,4,16]]}}}