{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T19:31:08Z","timestamp":1778700668603,"version":"3.51.4"},"reference-count":52,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2023,9,23]],"date-time":"2023-09-23T00:00:00Z","timestamp":1695427200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Doctoral School of the University of Medicine and Pharmacy of Craiova, Romania"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>As the population ages, a high prevalence of multimorbidity will affect the way physicians need to think about drug interactions. With microglia\u2019s important involvement in the pathology and progression of Alzheimer\u2019s disease (AD), understanding whether systemically administered drugs intended for other affections could impact microglia function, already impacted by the presence of beta-amyloid, is important. The aim of this study was to evaluate morphological changes of microglia, using in vivo 2-photon laser scanning microscopy, in a murine model of AD under systemic administration of sodium or calcium ion channel blockers in order to establish potential effects that these drugs might have on microglia under neuro-inflammatory conditions. A total of 30 mice (age 14\u201316 weeks, weight 20\u201325 g) were used, with 25 APP randomly divided into three groups. The remaining animals were CX3CR1GFP\/GFP male mice (n = 5) used as WT controls. After baseline behavior testing, all animals received daily intraperitoneal injections for 30 days according to the assigned group [WT (n = 5), Control (n = 5), Carbamazepine (n = 10), and Verapamil (n = 10)]. The results showed that the Verapamil treatment improved short-term memory and enhanced exploratory behavior in APP mice. The Carbamazepine treatment also improved short-term memory but did not elicit significant changes in anxiety-related behavior. Both Verapamil and Carbamazepine reduced the surveillance speed of microglia processes and changed microglia morphology in the cortex compared to the Control group. Due to their complex molecular machinery, microglia are potentially affected by drugs that do not target them specifically, and, as such, investigating these interactions could prove beneficial in our management of neurodegenerative pathologies.<\/jats:p>","DOI":"10.3390\/ijms241914474","type":"journal-article","created":{"date-parts":[[2023,9,24]],"date-time":"2023-09-24T10:36:26Z","timestamp":1695551786000},"page":"14474","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Chronic Administration of Ion Channel Blockers Impact Microglia Morphology and Function in a Murine Model of Alzheimer\u2019s Disease"],"prefix":"10.3390","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8516-9766","authenticated-orcid":false,"given":"Ianis Kevyn Stefan","family":"Boboc","sequence":"first","affiliation":[{"name":"Department of Pharmacology, University of Medicine and Pharmacy of Craiova, 200349 Craiova, Romania"},{"name":"Experimental Research Centre for Normal and Pathological Aging, University of Medicine and Pharmacy of Craiova, 200349 Craiova, Romania"},{"name":"U.M.F. Doctoral School Craiova, University of Medicine and Pharmacy of Craiova, 200349 Craiova, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexandru","family":"Cojocaru","sequence":"additional","affiliation":[{"name":"Experimental Research Centre for Normal and Pathological Aging, University of Medicine and Pharmacy of Craiova, 200349 Craiova, Romania"},{"name":"Department of Physiology, University of Medicine and Pharmacy of Craiova, 200349 Craiova, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gabriel","family":"Nedelea","sequence":"additional","affiliation":[{"name":"Experimental Research Centre for Normal and Pathological Aging, University of Medicine and Pharmacy of Craiova, 200349 Craiova, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5706-8722","authenticated-orcid":false,"given":"Bogdan","family":"Catalin","sequence":"additional","affiliation":[{"name":"Experimental Research Centre for Normal and Pathological Aging, University of Medicine and Pharmacy of Craiova, 200349 Craiova, Romania"},{"name":"Department of Physiology, University of Medicine and Pharmacy of Craiova, 200349 Craiova, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8089-1787","authenticated-orcid":false,"given":"Maria","family":"Bogdan","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of Medicine and Pharmacy of Craiova, 200349 Craiova, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1523-9116","authenticated-orcid":false,"given":"Daniela","family":"Calina","sequence":"additional","affiliation":[{"name":"Department of Clinical Pharmacy, University of Medicine and Pharmacy of Craiova, 200349 Craiova, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,9,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1093\/gerona\/glv128","article-title":"Global Multimorbidity Patterns: A Cross-Sectional, Population-Based, Multi-Country Study","volume":"71","author":"Garin","year":"2016","journal-title":"J. Gerontol. A Biol. Sci. Med. Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1437","DOI":"10.1007\/s11096-021-01308-0","article-title":"Management of Drug-Disease Interactions: A Best Practice from the Netherlands","volume":"43","author":"Diesveld","year":"2021","journal-title":"Int. J. Clin. Pharm."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"773","DOI":"10.3233\/JAD-170786","article-title":"Medical Comorbidity in Alzheimer\u2019s Disease: A Nested Case-Control Study","volume":"63","author":"Wang","year":"2018","journal-title":"J. Alzheimers Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"17294","DOI":"10.1074\/jbc.M500997200","article-title":"Calcium Dysregulation and Membrane Disruption as a Ubiquitous Neurotoxic Mechanism of Soluble Amyloid Oligomers","volume":"280","author":"Demuro","year":"2005","journal-title":"J. Biol. Chem."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"3348","DOI":"10.1073\/pnas.0813355106","article-title":"Small Molecule Blockers of the Alzheimer Abeta Calcium Channel Potently Protect Neurons from Abeta Cytotoxicity","volume":"106","author":"Diaz","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_6","first-page":"263","article-title":"Different Age Related Neurological and Cardiac Effects of Verapamil on a Transgenic Mouse Model of Alzheimer\u2019s Disease","volume":"47","author":"Cojocaru","year":"2021","journal-title":"Curr. Health Sci. J."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1016\/j.neuropharm.2010.10.002","article-title":"Verapamil Protects Dopaminergic Neuron Damage through a Novel Anti-Inflammatory Mechanism by Inhibition of Microglial Activation","volume":"60","author":"Liu","year":"2011","journal-title":"Neuropharmacology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1186\/s10020-022-00564-8","article-title":"Chronotherapeutic Neuroprotective Effect of Verapamil against Lipopolysaccharide-Induced Neuroinflammation in Mice through Modulation of Calcium-Dependent Genes","volume":"28","author":"Mosalam","year":"2022","journal-title":"Mol. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"815347","DOI":"10.3389\/fnagi.2022.815347","article-title":"Microglia Polarization From M1 to M2 in Neurodegenerative Diseases","volume":"14","author":"Guo","year":"2022","journal-title":"Front. Aging Neurosci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1159\/000519974","article-title":"Homoharringtonine Inhibits Alzheimer\u2019s Disease Progression by Reducing Neuroinflammation via STAT3 Signaling in APP\/PS1 Mice","volume":"21","author":"Jiang","year":"2022","journal-title":"Neurodegener. Dis."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1111\/j.1532-5415.2008.02089.x","article-title":"Chronic Pain and Obesity in Elderly People: Results from the Einstein Aging Study","volume":"57","author":"Mccarthy","year":"2009","journal-title":"J. Am. Geriatr. Soc."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Nakai, Y., Makizako, H., Kiyama, R., Tomioka, K., Taniguchi, Y., Kubozono, T., Takenaka, T., and Ohishi, M. (2019). Association between Chronic Pain and Physical Frailty in Community-Dwelling Older Adults. Int. J. Environ. Res. Public Health, 16.","DOI":"10.3390\/ijerph16081330"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/S0140-6736(16)31919-5","article-title":"Worldwide Trends in Blood Pressure from 1975 to 2015: A Pooled Analysis of 1479 Population-Based Measurement Studies with 19\u00b71 Million Participants","volume":"389","author":"Zhou","year":"2017","journal-title":"Lancet"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"872","DOI":"10.1093\/ije\/dyy016","article-title":"Contributions of Mean and Shape of Blood Pressure Distribution to Worldwide Trends and Variations in Raised Blood Pressure: A Pooled Analysis of 1018 Population-Based Measurement Studies with 88.6 Million Participants","volume":"47","author":"Ezzati","year":"2018","journal-title":"Int. J. Epidemiol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Cojocaru, A., Burada, E., B\u0103l\u0219eanu, A.T., Deftu, A.F., C\u0103t\u0103lin, B., Popa-Wagner, A., and Osiac, E. (2021). Roles of Microglial Ion Channel in Neurodegenerative Diseases. J. Clin. Med., 10.","DOI":"10.3390\/jcm10061239"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1038\/s41582-020-00435-y","article-title":"Neuroinflammation and Microglial Activation in Alzheimer Disease: Where Do We Go from Here?","volume":"17","author":"Leng","year":"2020","journal-title":"Nat. Rev. Neurol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Boboc, I.K.S., Rotaru-Zavaleanu, A.D., Calina, D., Albu, C.V., Catalin, B., and Turcu-Stiolica, A. (2023). A Preclinical Systematic Review and Meta-Analysis of Behavior Testing in Mice Models of Ischemic Stroke. Life, 13.","DOI":"10.3390\/life13020567"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1314","DOI":"10.1126\/science.1110647","article-title":"Neuroscience: Resting Microglial Cells Are Highly Dynamic Surveillants of Brain Parenchyma in Vivo","volume":"308","author":"Nimmerjahn","year":"2005","journal-title":"Science (80-)"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2000","DOI":"10.1038\/s41380-019-0407-3","article-title":"A Novel Role for the Late-Onset Alzheimer\u2019s Disease (LOAD)-Associated Protein Bin1 in Regulating Postsynaptic Trafficking and Glutamatergic Signaling","volume":"25","author":"Bermingham","year":"2020","journal-title":"Mol. Psychiatry"},{"key":"ref_20","first-page":"7","article-title":"Microglia Morphology in the Physiological and Diseased Brain\u2014From Fixed Tissue to in Vivo Conditions","volume":"59","author":"Stopper","year":"2018","journal-title":"Rom. J. Morphol. Embryol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"13211","DOI":"10.1038\/s41598-017-13581-z","article-title":"Quantitative Microglia Analyses Reveal Diverse Morphologic Responses in the Rat Cortex after Diffuse Brain Injury","volume":"7","author":"Morrison","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1113","DOI":"10.1002\/dvdy.582","article-title":"Microglial Morphology in the Somatosensory Cortex across Lifespan. A Quantitative Study","volume":"252","author":"Godeanu","year":"2023","journal-title":"Dev. Dyn."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1038\/s41380-019-0583-1","article-title":"Long-Read Sequencing Reveals the Complex Splicing Profile of the Psychiatric Risk Gene CACNA1C in Human Brain","volume":"25","author":"Clark","year":"2020","journal-title":"Mol. Psychiatry"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1461","DOI":"10.1016\/j.molimm.2005.01.014","article-title":"Molecular Characterization of L-Type Calcium Channel Splice Variants Expressed in Human T Lymphocytes","volume":"42","author":"Kotturi","year":"2005","journal-title":"Mol. Immunol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1002\/jnr.24585","article-title":"Targeting Microglia L-Type Voltage-Dependent Calcium Channels for the Treatment of Central Nervous System Disorders","volume":"99","author":"Hopp","year":"2021","journal-title":"J. Neurosci. Res."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Zhang, Y., Zhang, L., Zhou, C.-N., Jiang, L., Tang, J., Liang, X., Fan, J.-H., Dou, X.-Y., and Tang, Y. (2020). Oligodendrocyte Generation and Maturation in the Hippocampus May Be a Target of Fluoxetine for Delaying Cognitive Dysfunction during Early Alzheimer\u2019s Disease. arXiv.","DOI":"10.21203\/rs.2.24601\/v1"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"bay050","DOI":"10.1093\/database\/bay050","article-title":"A Systematic Approach for Identifying Shared Mechanisms in Epilepsy and Its Comorbidities","volume":"2018","author":"Hoyt","year":"2018","journal-title":"Database"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"964077","DOI":"10.3389\/fnagi.2022.964077","article-title":"Antarctic Krill Oil Exhibited Synergistic Effects with Nobiletin and Theanine in Ameliorating Memory and Cognitive Deficiency in SAMP8 Mice: Applying the Perspective of the Sea\u2013Land Combination to Retard Brain Aging","volume":"14","author":"Wang","year":"2022","journal-title":"Front. Aging Neurosci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"763866","DOI":"10.3389\/fphar.2021.763866","article-title":"TRPV1-Mediated Microglial Autophagy Attenuates Alzheimer\u2019s Disease-Associated Pathology and Cognitive Decline","volume":"12","author":"Wang","year":"2022","journal-title":"Front. Pharmacol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"940","DOI":"10.1038\/sj.embor.7400784","article-title":"A\u03b242-Driven Cerebral Amyloidosis in Transgenic Mice Reveals Early and Robust Pathology","volume":"7","author":"Radde","year":"2006","journal-title":"EMBO Rep."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"(1988). Verapamil Appears to Reduce Anxiety and Panic Attacks. InPharma, 637, 12.","DOI":"10.1007\/BF03308285"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"479","DOI":"10.1007\/978-1-4613-2363-1_75","article-title":"Carbamazepine: Treatment of Affective Illness and Anxiety Syndromes","volume":"17","author":"Uhde","year":"1985","journal-title":"Psychiatry State Art"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1089\/adt.2013.552","article-title":"Evaluating State Dependence and Subtype Selectivity of Calcium Channel Modulators in Automated Electrophysiology Assays","volume":"12","author":"Kuryshev","year":"2014","journal-title":"Assay Drug Dev. Technol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1002","DOI":"10.1016\/j.neurobiolaging.2013.10.090","article-title":"Age-Related Downregulation of the CaV3.1 T-Type Calcium Channel as a Mediator of Amyloid Beta Production","volume":"35","author":"Rice","year":"2014","journal-title":"Neurobiol. Aging"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"805755","DOI":"10.3389\/fncel.2021.805755","article-title":"Pharmacological Investigations in Glia Culture Model of Inflammation","volume":"15","author":"Ismail","year":"2021","journal-title":"Front. Cell. Neurosci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"624","DOI":"10.1038\/nrneurol.2012.200","article-title":"Neuroprotection and Repair in Multiple Sclerosis","volume":"8","author":"Franklin","year":"2012","journal-title":"Nat. Rev. Neurol."},{"key":"ref_37","first-page":"556","article-title":"Aripiprazole Lauroxil (Aristada): An Extended-Release, Long-Acting Injection For the Treatment of Schizophrenia","volume":"41","author":"Cruz","year":"2016","journal-title":"Pharm. Ther."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"310","DOI":"10.2174\/138920008784220664","article-title":"Drugs Behave as Substrates, Inhibitors and Inducers of Human Cytochrome P450 3A4","volume":"9","author":"Zhou","year":"2008","journal-title":"Curr. Drug Metab."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"367","DOI":"10.4070\/kcj.2009.39.9.367","article-title":"Prognosis and Natural History of Drug-Related Bradycardia","volume":"39","author":"Jang","year":"2009","journal-title":"Korean Circ. J."},{"key":"ref_40","unstructured":"(2023, February 10). FDA 2002 Highlights of Prescribing Information Abilify\u00ae (Aripiprazole), Available online: https:\/\/www.accessdata.fda.gov\/drugsatfda_docs\/label\/2014\/021436s038,021713s030,021729s022,021866s023lbl.pdf."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1018","DOI":"10.4103\/1673-5374.324843","article-title":"Repurposing Verapamil for Prevention of Cognitive Decline in Sporadic Alzheimer\u2019s Disease","volume":"17","author":"Ahmed","year":"2022","journal-title":"Neural. Regen. Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"4106","DOI":"10.1128\/MCB.20.11.4106-4114.2000","article-title":"Analysis of Fractalkine Receptor CX(3)CR1 Function by Targeted Deletion and Green Fluorescent Protein Reporter Gene Insertion","volume":"20","author":"Jung","year":"2000","journal-title":"Mol. Cell. Biol."},{"key":"ref_43","first-page":"55718","article-title":"Novel Object Recognition Test for the Investigation of Learning and Memory in Mice","volume":"2017","author":"Lueptow","year":"2017","journal-title":"J. Vis. Exp."},{"key":"ref_44","first-page":"151","article-title":"Comparative Aspects of Microglia Reaction in White and Gray Matter","volume":"39","author":"Mitran","year":"2013","journal-title":"Curr. Health Sci. J."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1038\/nmeth706","article-title":"Sulforhodamine 101 as a Specific Marker of Astroglia in the Neocortex in Vivo","volume":"1","author":"Nimmerjahn","year":"2004","journal-title":"Nat. Methods"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1023\/B:VISI.0000029664.99615.94","article-title":"Distinctive Image Features from Scale-Invariant Keypoints","volume":"60","author":"Lowe","year":"2004","journal-title":"Int. J. Comput. Vis."},{"key":"ref_47","unstructured":"(2023, August 10). Registration. Available online: https:\/\/imagej.net\/imaging\/registration."},{"key":"ref_48","first-page":"1","article-title":"Original Paper Microglia Branching Using a Sholl Analysis Method","volume":"39","author":"Alexandru","year":"2013","journal-title":"Curr. Health Sci. J."},{"key":"ref_49","unstructured":"(2023, August 10). Sholl Analysis. Available online: https:\/\/imagej.net\/plugins\/sholl-analysis."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Jung, C.K.E., Keppler, K., Steinbach, S., Blazquez-Llorca, L., and Herms, J. (2015). Fibrillar Amyloid Plaque Formation Precedes Microglial Activation. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0119768"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"586","DOI":"10.1038\/s41590-021-00913-5","article-title":"Microglia Use TAM Receptors to Detect and Engulf Amyloid \u03b2 Plaques","volume":"22","author":"Huang","year":"2021","journal-title":"Nat. Immunol."},{"key":"ref_52","first-page":"379872","article-title":"In Vivo Imaging of Microglia with Multiphoton Microscopy","volume":"10","author":"Bacskai","year":"2018","journal-title":"Front. Aging Neurosci."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/19\/14474\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T20:56:34Z","timestamp":1760129794000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/19\/14474"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,9,23]]},"references-count":52,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2023,10]]}},"alternative-id":["ijms241914474"],"URL":"https:\/\/doi.org\/10.3390\/ijms241914474","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,9,23]]}}}