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After the withdrawal of sensory reflexes, mice were shaved on the back of the neck to the base of the skull and then disinfected with ethanol wipes. The mouse head was stabilized on a stereotaxic frame (David Kopf Instruments, Tujunga, CA, USA) without fixing the jaw on the tooth adapter. No lateral head movement ensured proper head fixation on the stereotactic frame. To level the mouse on the stereotaxic frame, the body was placed on a custom-made box. A linear skin incision was made in the midline from the top of the skull to the neck using a scalpel blade (No.11). The skin was retracted, and the muscles were exposed after the removal of the superficial layer of connective tissue using forceps. Using fine scissors muscles were detached from the base of the occipital bone. Extra muscles and blood were removed using a cotton tip, and the area was cleaned for better visualization of the Cisterna Magna. Little connective tissue was left on the top of the dura that stabilized the Hamilton needle and prevented the backflow of the CSF. Hemostasis was prevented during the entire process, either using a cotton swab or a suction aspirator. A Hamilton syringe containing a 30 G needle was attached to a micro syringe injector pump (KD Scientific) and placed on a stereotaxic frame. The Hamilton syringe needle was adjusted at a 60-degree angle. Mice heads were pressed down using a tooth bar adaptor to expose the cisterna magna syringe injector needle was aligned with the cisterna magna linearly, and the needle was inserted into the cisterna magna after puncturing the dura. CSF was drawn into the syringe by slow and smooth aspiration by starting the microinjector pump aspirator at a flow rate of 8 µl/min. CSF was inspected with the naked eye for quality and clarity. The CSF was transferred into a new 500 µl centrifuge tube and briefly spun down, and the supernatant was transferred into a new vial and snap frozen on dry ice for further analysis. Tissue collection   After CSF collections, mice were transcardially perfused with ice-cold PBS (1X). Mice brains were immediately removed by dissection without damaging the brain structure. The right hemisphere of the brain was snap frozen on dry ice either intact or after regional dissection (cerebellum, brain stem, cortex, hippocampus, and striatum/thalamus) for downstream proteomics analysis, while the left hemisphere was post-fixed in 4% PFA for 24 hours (Cat# J19943.K2, Thermo Scientific Fisher) and then transferred to the 30% sucrose for immunohistological studies. Tissue processing for protein-based analysis, including Western blot Protein digestion MS, Protein identification, and quantification     "],"repository":["Pride"],"quantification_method":[""],"modification":[""],"data_protocol":["Protein filtering, imputation, and normalization LC-MS/MS data analysis of 5xFAD Mouse CSF      Proteomic data from 5xFAD mouse CSF were first process to remove the blood contamination across the samples using regression analysis and variance partitiona analysis after that it processed in Perseus (http://www.perseus-framework.org/). Proteins were retained if at least 50% of values were present in one group, and intensities were log2-transformed. Missing values were imputed in Perseus by random sampling from a normal distribution. To correct sample loading differences, column-sum median normalization was performed in R, where total protein intensity was calculated per sample, and each was scaled by the ratio of the global median column sum to its own sum. Group comparisons were assessed using one-way ANOVA and two-sample t-tests, with statistical significance defined at p < 0.05. Differentially expressed proteins were visualized using volcano plots generated in ggplot2, displaying log2 fold change against P-value.        "],"omics_type":["Proteomics"],"labhead":["upasna srivastava"],"instrument_platform":[""],"submission_type":["PARTIAL"],"labhead_affiliation":["Department of Neurology, Yale School of Medicine, Yale University, new haven CT, USA"],"species":["Mus Musculus (mouse)"],"publication":["Not available"],"submitter_mail":["upasna.srivastava@yale.edu"],"submitter_affiliation":["YALE UNIVERSITY"],"submitter_country":["United States"],"additional_accession":[]},"is_claimable":false,"name":"Kv1.3 inhibition alleviates neuropathology via neuroinflammatory and resilience pathways in a mouse model of Aβ pathology","description":"Kv1.3, a voltage-gated potassium channel, is a therapeutic target in various neuropathological conditions. In rodent models, Kv1.3 inhibitors have been shown to inhibit disease-induced neuroinflammation. However, the cellular mechanisms of disease-modification by Kv1.3 blockers and the signaling pathways regulated by Kv1.3 and are unclear. In this -study, we aim to delineate the major protective mechanisms of Kv1.3 blockade in Alzheimer’s disease (AD) pathology by investigating the brain-derived bulk and single nuclear transcriptomes (snRNAseq) and bulk proteomes, along with neurobehavioral and pathological changes in 5xFAD mice. Following 3 months of Kv1.3 blockade via either small molecule PAP-1 or ShK-223 peptide, we observed Ab plaque reduction with both drugs and improved behavior with PAP-1. snRNAseq of brain nuclei identified neuronal and glial populations with expected AD-related changes in microglia. Pseudo-bulk analyses showed that PAP-1 disproportionately impacted oligodendrocytes and microglia while ShK-223 had more pronounced effects on glutamatergic neurons. PAP-1 also increased expression of myelination genes in oligodendrocytes and synaptic genes in neurons, with distinct effects on excitatory and inhibitory neurons. Cell-cell communication analyses suggested that PAP-1 increased crosstalk between neurons and astrocytes with endothelial cells. Bulk brain transcriptomics and proteomics with PAP-1 showed increased levels of synaptic and cognitive resilience-associated proteins while decreasing glial activation and interferon (IFN) responsive genes/proteins. PAP-1 also decreased STAT1 phosphorylation in the 5xFAD brain. To gain further mechanistic insight into Kv1.3's role in interferon signaling in microglia, we confirmed physical interactions between Kv1.3 and STAT1 and STAT3 in BV2 microglia overexpressing Kv1.3. We also found that Kv1.3 blockade (PAP-1) specifically reduced type 2 IFN but not type 1 IFN-induced phosphorylation of STAT1/3, a finding with further validated in another microglial cell line as well as in primary microglia derived from wild-type or Kv1.3-knockout mice. Lastly, we identified novel proteomic changes occurring in cerebrospinal fluid that reflect disease-modification by Kv1.3 blockers. This study reveals cell type-specific effects of Kv1.3 blockers in AD pathology that correspond with reduced neuropathology and neuroinflammation, augmentation of resilience pathways and neuro-vascular coupling and translationally relevant biofluid biomarkers of therapeutic effect. We also provide novel mechanistic insights into functional coupling between Kv1.3 channels and type-2 IFN signaling pathways in microglia.","dates":{"publication":"2026-07-13","submission":"2025-12-16"},"accession":"PXD072055","cross_references":{"TAXONOMY":["NEWT:68239","NEWT:108018","NEWT:377960","NEWT:1131","NEWT:2042546","NEWT:295546","NEWT:258594","NEWT:1126","NEWT:1129","NEWT:309807","NEWT:89184","NEWT:309800","NEWT:1323429","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:2315688","NEWT:592321","NEWT:1073882","NEWT:6819","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:10034","NEWT:661410","NEWT:224326","NEWT:376619","NEWT:3765","NEWT:3767","NEWT:433727","NEWT:135622","NEWT:672127","NEWT:656061","NEWT:37965","NEWT:656064","NEWT:3760","NEWT:36630","NEWT:381198","NEWT:418106","NEWT:10029","NEWT:913645","NEWT:2492962","NEWT:641809","NEWT:44250","NEWT:317447","NEWT:5530","NEWT:868565","NEWT:135674","NEWT:1562698","NEWT:12637","NEWT:2164133","NEWT:2767846","NEWT:314344","NEWT:108061","NEWT:224308","NEWT:49928","NEWT:160621","NEWT:110368","NEWT:1916073","NEWT:1310161","NEWT:1230383","NEWT:77133","NEWT:1310165","NEWT:114727","NEWT:5516","NEWT:498211","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:1310173","NEWT:37919","NEWT:161934","NEWT:156587","NEWT:1148","NEWT:5508","NEWT:419481","NEWT:5507","NEWT:1140","NCBITaxon:2157","NEWT:2066563","NEWT:1143","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:709964","NEWT:3712","NEWT:3711","NEWT:3714","NEWT:3715","NEWT:2065263","NEWT:1549675","NEWT:177437","NEWT:9014","NEWT:10891","NEWT:226126","NEWT:1616117","NEWT:34865","NEWT:28174","NEWT:550537","NEWT:284812","NEWT:3701","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:1246791","NEWT:9005","NEWT:34871","NEWT:9006","NEWT:33548","NEWT:1883426","NEWT:657321","NEWT:10886","NEWT:3708","NEWT:536231","NEWT:219813","NEWT:215451","NEWT:460519","NEWT:272844","NEWT:1348799","NEWT:55784","NEWT:485","NEWT:2709072","NEWT:2499525","NEWT:142786","NEWT:2853422","NEWT:487","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:46835","NEWT:216778","NEWT:376686","NEWT:237610","EFO:0001352","NEWT:219809","NEWT:219808","NEWT:119562","NEWT:1883446","NEWT:1233435","NEWT:118223","NEWT:141469","NEWT:987059","NEWT:491","NEWT:460523","NEWT:85694","NEWT:85692","NEWT:3750","NEWT:291146","NEWT:29466","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:278161","NEWT:415978","NEWT:410289","NEWT:191701","NEWT:9054","NEWT:472","NEWT:29459","NEWT:1380096","NEWT:28128","NEWT:470","NEWT:41122","NEWT:1328300","NEWT:748693","NCBITaxon:50557","NEWT:29491","NEWT:911022","NEWT:446","NEWT:153481","NEWT:445","NEWT:2003758","NEWT:63366","NEWT:63367","NEWT:29488","NEWT:215402","NEWT:3726","NEWT:30455","NEWT:28141","NEWT:9031","NEWT:3728","NEWT:108931","NEWT:219799","NEWT:219798","NEWT:548","NEWT:150475","NEWT:256737","NEWT:51350","NEWT:8694","NEWT:8692","NEWT:542","NEWT:394503","NEWT:13076","NEWT:219785","NEWT:451515","NEWT:8673","NEWT:550","NEWT:10804","NEWT:554","NEWT:451516","NEWT:552","NEWT:1325291","NEWT:42410","NEWT:1225786","NEWT:771870","NEWT:51338","NEWT:9995","NEWT:28112","NEWT:49012","NEWT:28110","NEWT:10832","NEWT:507601","NEWT:520","NEWT:64657","NEWT:186441","NEWT:2585209","NEWT:643680","NEWT:214092","NEWT:40697","NEWT:13095","NEWT:162425","NEWT:242124","NEWT:9986","NEWT:8654","NEWT:8658","NEWT:1268063","NEWT:8657","NEWT:9983","NEWT:8655","NEWT:595495","NEWT:80833","NEWT:479096","NEWT:28108","NEWT:28104","NEWT:1114957","NEWT:568703","NEWT:568708","NEWT:52641","NEWT:353152","NEWT:353153","NEWT:227377","NEWT:1774284","NEWT:27689","NEWT:207769","NEWT:40674","NEWT:51329","NCBITaxon:1763","NEWT:443906","NEWT:13037","NEWT:519","NEWT:7394","NEWT:6063","NEWT:404330","NEWT:1328388","NEWT:300641","NEWT:39655","NEWT:38323","NEWT:206411","NEWT:80869","NEWT:214053","NEWT:80863","NEWT:665890","NEWT:52639","NEWT:52638","NEWT:8697","NEWT:861109","NEWT:8695","NEWT:1196866","NEWT:510516","NEWT:1045775","NEWT:1123869","NCBITaxon:1335626","NEWT:7370","NEWT:83906","NEWT:99488","NEWT:1134506","NEWT:255470","NEWT:27697","NEWT:86185","NEWT:9935","NEWT:287889","NEWT:1028307","NEWT:52603","NEWT:92867","NEWT:92866","NEWT:1344414","NEWT:202950","NEWT:741158","NEWT:1293091","NEWT:202956","NEWT:315750","NEWT:632957","NEWT:9925","NCBITaxon:9606","NEWT:1436183","NEWT:4232","NEWT:9922","NEWT:416348","NEWT:11298","NEWT:4236","NEWT:398031","NEWT:196627","NEWT:523840","NEWT:1076878","NEWT:200308","NEWT:200302","NEWT:9913","NEWT:9915","NEWT:105841","NEWT:1170227","NEWT:2666255","NEWT:999810","NEWT:28995","NEWT:1392998","NEWT:1150621","NEWT:1085791","NEWT:745531","NEWT:226900","NEWT:231490","NEWT:590651","NEWT:449239","NEWT:244704","NEWT:9909","NEWT:9902","NEWT:430615","NEWT:563041","NEWT:36599","NEWT:41949","NEWT:9900","NEWT:326423","NEWT:326424","NEWT:13013","NEWT:749906","NEWT:418985","NEWT:749907","NEWT:431947","NEWT:142809","NEWT:1095183","NEWT:302409","NEWT:9979","NEWT:130821","NEWT:2320211","NEWT:27606","NEWT:290610","NEWT:1519788","NEWT:59202","NEWT:9975","NEWT:8643","NEWT:1801631","NEWT:1159899","NEWT:502780","NEWT:860688","NEWT: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