{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T15:53:45Z","timestamp":1785426825561,"version":"3.56.0"},"posted":{"date-parts":[[2026,4,17]]},"group-title":"Neuroscience","reference-count":40,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,4,17]],"date-time":"2026-04-17T00:00:00Z","timestamp":1776384000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"award":["R35GM142712"],"award-info":[{"award-number":["R35GM142712"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["1RF1AG066905"],"award-info":[{"award-number":["1RF1AG066905"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["5R01MH063352"],"award-info":[{"award-number":["5R01MH063352"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R01GM144377"],"award-info":[{"award-number":["R01GM144377"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R01GM151556"],"award-info":[{"award-number":["R01GM151556"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,4,17]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Introduction<\/jats:title>\n                  <jats:p>\n                    Here, we create a conditional\n                    <jats:italic>Adra2a<\/jats:italic>\n                    line and use it to show that sedative, hypnotic, and hypothermic effects of \u03b1\n                    <jats:sub>2<\/jats:sub>\n                    -agonists are neuronally mediated via the \u03b1\n                    <jats:sub>2A<\/jats:sub>\n                    adrenergic receptor.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>\n                    We generated mice with loxP sites flanking\n                    <jats:italic>Adra2a<\/jats:italic>\n                    using CRISPR\/Cas9 gene targeting. This line was crossed with lines encoding Cre recombinase (Cre) under the control of the Vgat, Snap25, and Dbh promoters. Cell-specific knockout was confirmed using fluorescent in-situ hybridization demonstrating targeted reduction in\n                    <jats:italic>Adra2a<\/jats:italic>\n                    mRNA in the appropriate cell types. Mice were given intraperitoneal dexmedetomidine (0.3 or 1 mg\/kg) or saline, and 20 minutes later righting reflex was assessed, followed by 3 rounds of rotarod testing, with fall time and end temperature recorded. Spontaneous activity was recorded using beam break for an hour after. Mice of each genotype were implanted with EEG leads and recorded while given 0.3 mg\/kg IP dexmedetomidine.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>\n                    We created a conditional knockout and demonstrated cell-type specific reduction of\n                    <jats:italic>Adra2a<\/jats:italic>\n                    mRNA in crossed lines with cell-specific Cre. The pan-neuronal\n                    <jats:italic>Adra2a<\/jats:italic>\n                    knockout showed resistance to all temperature, sedative, and hypnotic effect endpoints in response to the \u03b1\n                    <jats:sub>2<\/jats:sub>\n                    -agonist dexmedetomidine. Adrenergic knockout demonstrated resistance to \u03b1\n                    <jats:sub>2<\/jats:sub>\n                    -agonist hypnosis and moderate resistance to hypothermia and impaired coordination with forced movement. GABAergic knockout showed resistance only to impairment of spontaneous movement by \u03b1\n                    <jats:sub>2<\/jats:sub>\n                    -agonists. Spectral analysis of the EEG showed an increase in proportion of delta power with a sedative dose of dexmedetomidine in all lines except the pan-neuronal\n                    <jats:italic>Adra2a<\/jats:italic>\n                    knockout.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Discussion<\/jats:title>\n                  <jats:p>\n                    Future studies will pursue both the specific subtype(s) and location of neuronal populations responsible for sedative, hypnotic, and hypothermic effects of \u03b1\n                    <jats:sub>2<\/jats:sub>\n                    -agonists.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.04.14.712766","type":"posted-content","created":{"date-parts":[[2026,4,17]],"date-time":"2026-04-17T17:35:18Z","timestamp":1776447318000},"source":"Crossref","is-referenced-by-count":0,"title":["A Novel Conditional Adra2a-Knockout Mouse Line Reveals Cell-specific Contributions to Specific Dimensions of Sedation"],"prefix":"10.64898","author":[{"given":"Noah L.","family":"Fryou","sequence":"first","affiliation":[{"name":"Department of Anesthesiology and Critical Care, University of Pennsylvania Perelman School of Medicine, Philadelphia, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Timothy","family":"Jiang","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology and Critical Care, University of Pennsylvania Perelman School of Medicine, Philadelphia, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nathan","family":"Frick","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology and Critical Care, University of Pennsylvania Perelman School of Medicine, Philadelphia, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paula","family":"Kwasniewska","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology and Critical Care, University of Pennsylvania Perelman School of Medicine, Philadelphia, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mikhail Y.","family":"Lipin","sequence":"additional","affiliation":[{"name":"Department of Systems Pharmacology and Translational Therapeutics, University of Pennsylvania Perelman School of Medicine, Philadelphia, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2803-6078","authenticated-orcid":false,"given":"Max B.","family":"Kelz","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology and Critical Care, University of Pennsylvania Perelman School of Medicine, Philadelphia, USA"},{"name":"Center for the Neuroscience of Unconsciousness and Reanimation Research Alliance, University of Pennsylvania Perelman School of Medicine, Philadelphia, USA"},{"name":"Chronobiology and Sleep Institute, University of Pennsylvania Perelman School of Medicine, Philadelphia, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steven A.","family":"Thomas","sequence":"additional","affiliation":[{"name":"Department of Systems Pharmacology and Translational Therapeutics, University of Pennsylvania Perelman School of Medicine, Philadelphia, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7078-4603","authenticated-orcid":false,"given":"Andrew","family":"McKinstry-Wu","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology and Critical Care, University of Pennsylvania Perelman School of Medicine, Philadelphia, USA"},{"name":"Center for the Neuroscience of Unconsciousness and Reanimation Research Alliance, University of Pennsylvania Perelman School of Medicine, Philadelphia, USA"},{"name":"Chronobiology and Sleep Institute, University of Pennsylvania Perelman School of Medicine, Philadelphia, 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