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GlyRs are clustered in the postsynaptic membrane by strong interactions of the \u03b2 subunit with the scaffold protein gephyrin. Even though\n                    <jats:italic>Glrb<\/jats:italic>\n                    mRNA is highly expressed throughout the brain, the existence of synaptic GlyRs remains controversial as there is little conclusive evidence using conventional fluorescence microscopy and electrophysiological recordings. Here, we exploit the high sensitivity and spatial resolution of single molecule localisation microscopy (SMLM) to investigate the presence of GlyRs at inhibitory synapses in the brain, focusing on several areas of the telencephalon. Making use of a knock-in mouse model expressing endogenous mEos4b-tagged GlyR\u03b2, we identified few GlyRs in sub-regions of the hippocampus. Dual-colour SMLM revealed that these sparse receptors are integrated within the postsynaptic gephyrin domain, pointing to a possible role in maintaining the structural integrity of inhibitory synapses. In contrast, we found functionally relevant numbers of synaptic GlyRs at inhibitory synapses in the ventral striatum. Our results highlight the strength of SMLM to detect few and sparsely distributed synaptic molecules in complex samples and to analyse their organisation with high spatial precision.\n                  <\/jats:p>","DOI":"10.7554\/elife.109447","type":"journal-article","created":{"date-parts":[[2025,12,15]],"date-time":"2025-12-15T15:30:41Z","timestamp":1765812641000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.109447","source":"Crossref","is-referenced-by-count":4,"title":["Single molecule counting detects low-copy glycine receptors in hippocampal and striatal synapses"],"prefix":"10.7554","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3116-6340","authenticated-orcid":true,"given":"Serena","family":"Camuso","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03xjwb503","id-type":"ROR","asserted-by":"publisher"}],"name":"Neuro-Bic\u00eatre, Inserm U1195, Universit\u00e9 Paris-Saclay","place":["Paris, 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Abadi","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03xjwb503","id-type":"ROR","asserted-by":"publisher"}],"name":"Neuro-Bic\u00eatre, Inserm U1195, Universit\u00e9 Paris-Saclay","place":["Paris, France"]}],"role":[{"role":"author","vocabulary":"crossref"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2069-8250","authenticated-orcid":true,"given":"Cl\u00e9mence","family":"Mille","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03xjwb503","id-type":"ROR","asserted-by":"publisher"}],"name":"Neuro-Bic\u00eatre, Inserm U1195, Universit\u00e9 Paris-Saclay","place":["Paris, 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