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The PreC hub appears the most resilient to AD pathology, suggesting differential vulnerability within the DMN. However, the mechanisms that underlie this differential pathobiology remain obscure. Here, we investigated changes in RNA polymerase II (RNA pol II) and splicing proteins U1\u201070K, U1A, SRSF2, and hnRNPA2B1, phosphorylated AT8 tau, 3R and 4Rtau isoforms containing neurons and amyloid plaques in layers III and V\u2013VI in FC, PreC, and PCC obtained from individuals with a preclinical diagnosis of no cognitive impairment (NCI), mild cognitive impairment (MCI), and mild\/moderate mAD. We found a significant increase in pS5\u2010RNA pol II levels in FC NCI, U1\u201070K in PreC MCI and mAD, and hnRNPA2B1 and SRSF2 levels in PCC mAD. 1N3Rtau levels were significantly increased in FC, decreased in PreC in mAD, and unchanged in PCC, whereas 1N4Rtau increased in mAD across the hubs. SRSF2, U1\u201070K, U1A, and hnRNPA2B1 nuclear optical density (OD), size, and number were unchanged across groups in FC and PCC, while PreC OD hnRNPA2B1 was significantly greater in mAD. Mislocalized U1A and U1\u201070K tangle\u2010like structures were found in a few PCC cases and colocalized with AT8\u2010bearing neurofibrillary tangles (NFTs). FC pS5\u2010RNA pol II, PreC U1\u201070K, Pre pS5,2\u2010RNA pol II, and PCC hnRNPA2B1 and SRSF2 protein levels were associated with cognitive decline but not neuropathology across clinical groups. By contrast, splicing protein nuclear OD measures, size, counts, and mislocalized U1\u201070K and U1A NFT\u2010like structures were not correlated with NFT or plaque density, cognitive domains, and neuropathological criteria in DMN hubs. Findings suggest that RNA splicing protein alterations and U1 mislocalization contribute differentially to DMN pathogenesis and cognitive deterioration in AD.<\/jats:p>","DOI":"10.1111\/bpa.70004","type":"journal-article","created":{"date-parts":[[2025,3,23]],"date-time":"2025-03-23T23:04:19Z","timestamp":1742771059000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Spliceosome protein alterations differentiate hubs of the default mode connectome during the progression of Alzheimer's disease"],"prefix":"10.1111","volume":"35","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-1412-5721","authenticated-orcid":false,"given":"Sylvia E.","family":"Perez","sequence":"first","affiliation":[{"name":"Department of Translational Neurosciences Barrow Neurological Institute  Phoenix Arizona 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