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In mouse primary visual cortex, excitatory neurons with similar response properties are more likely to be synaptically connected\n                    <jats:sup>1\u20138<\/jats:sup>\n                    ; however, broader connectivity rules remain unknown. Here we leverage the millimetre-scale MICrONS dataset to analyse synaptic connectivity and functional properties of neurons across cortical layers and areas. Our results reveal that neurons with similar response properties are preferentially connected within and across layers and areas\u2014including feedback connections\u2014supporting the universality of \u2018like-to-like\u2019 connectivity across the visual hierarchy. Using a validated digital twin model, we separated neuronal tuning into feature (what neurons respond to) and spatial (receptive field location) components. We found that only the feature component predicts fine-scale synaptic connections beyond what could be explained by the proximity of axons and dendrites. We also discovered a higher-order rule whereby postsynaptic neuron cohorts downstream of presynaptic cells show greater functional similarity than predicted by a pairwise like-to-like rule. Recurrent neural networks trained on a simple classification task develop connectivity patterns that mirror both pairwise and higher-order rules, with magnitudes similar to those in\u00a0MICrONS data. Ablation studies in these recurrent neural networks reveal that disrupting like-to-like connections impairs performance more than disrupting random connections. These findings suggest that these connectivity principles may have a functional role in sensory processing and learning, highlighting shared principles between biological and artificial systems.\n                  <\/jats:p>","DOI":"10.1038\/s41586-025-08840-3","type":"journal-article","created":{"date-parts":[[2025,4,9]],"date-time":"2025-04-09T11:03:23Z","timestamp":1744196603000},"page":"459-469","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":56,"title":["Functional connectomics reveals general wiring rule in mouse visual cortex"],"prefix":"10.1038","volume":"640","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8610-5182","authenticated-orcid":false,"given":"Zhuokun","family":"Ding","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul G.","family":"Fahey","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stelios","family":"Papadopoulos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1084-7059","authenticated-orcid":false,"given":"Eric Y.","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brendan","family":"Celii","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-0669-7968","authenticated-orcid":false,"given":"Christos","family":"Papadopoulos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andersen","family":"Chang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1949-9457","authenticated-orcid":false,"given":"Alexander B.","family":"Kunin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dat","family":"Tran","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiakun","family":"Fu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhiwei","family":"Ding","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Saumil","family":"Patel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-9158-8408","authenticated-orcid":false,"given":"Lydia","family":"Ntanavara","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-0260-9036","authenticated-orcid":false,"given":"Rachel","family":"Froebe","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kayla","family":"Ponder","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Taliah","family":"Muhammad","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J. 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