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However, our understanding of GABA\n                  <jats:sub>A<\/jats:sub>\n                  R pharmacology has been hindered by the vast number of pentameric assemblies that can be derived from a total 19 different subunits and the lack of structural knowledge of clinically relevant receptors. Here, we isolate native murine GABA\n                  <jats:sub>A<\/jats:sub>\n                  R assemblies containing the widely expressed \u03b1\n                  <jats:sub>1<\/jats:sub>\n                  subunit, and elucidate their structures in complex with drugs used to treat insomnia (zolpidem and flurazepam) and postpartum depression (the neurosteroid allopregnanolone). Using cryo-EM analysis and single-molecule photobleaching experiments, we uncover only three structural populations in the brain: the canonical \u03b1\n                  <jats:sub>1<\/jats:sub>\n                  \u03b22\u03b3\n                  <jats:sub>2<\/jats:sub>\n                  receptor containing two \u03b1\n                  <jats:sub>1<\/jats:sub>\n                  subunits and two unanticipated assemblies containing one \u03b1\n                  <jats:sub>1<\/jats:sub>\n                  and either an \u03b1\n                  <jats:sub>2<\/jats:sub>\n                  , \u03b1\n                  <jats:sub>3<\/jats:sub>\n                  or \u03b1\n                  <jats:sub>5<\/jats:sub>\n                  subunit. Both of the noncanonical assemblies feature a more compact arrangement between the transmembrane and extracellular domains. Interestingly, allopregnanolone is bound at the transmembrane \u03b1\/\u03b2 subunit interface, even when not added to the sample, revealing an important role for endogenous neurosteroids in modulating native GABA\n                  <jats:sub>A<\/jats:sub>\n                  Rs. Together with structurally engaged lipids, neurosteroids produce global conformational changes throughout the receptor that modify both the pore diameter and binding environments for GABA and insomnia medications. Together, our data reveal that GABA\n                  <jats:sub>A<\/jats:sub>\n                  R assembly is a strictly regulated process that yields a small number of structurally distinct complexes, defining a structural landscape from which subtype-specific drugs can be developed.\n                <\/jats:p>","DOI":"10.1101\/2023.02.16.528867","type":"posted-content","created":{"date-parts":[[2023,2,16]],"date-time":"2023-02-16T22:25:13Z","timestamp":1676586313000},"source":"Crossref","is-referenced-by-count":3,"title":["Regulated assembly and neurosteroid modulation constrain GABA\n                  <sub>A<\/sub>\n                  receptor pharmacology\n                  <i>in vivo<\/i>"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6070-5545","authenticated-orcid":false,"given":"Chang","family":"Sun","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hongtao","family":"Zhu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2805-2526","authenticated-orcid":false,"given":"Sarah","family":"Clark","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8549-2360","authenticated-orcid":false,"given":"Eric","family":"Gouaux","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2023022010200978000_2023.02.16.528867v1.1","doi-asserted-by":"crossref","unstructured":"Lee, V. & Maguire, J . 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