{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T07:16:44Z","timestamp":1788419804448,"version":"build-2803163510"},"reference-count":45,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T00:00:00Z","timestamp":1788393600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Neurosci."],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>We previously demonstrated that apolipoprotein E (apoE4) accelerates neurodegeneration in a transgenic mouse model, but the mechanism is still unclear. A previous study reported that death-associated protein kinase 1 (DAPK1) interacts with the NR2B subunit of the N-methyl-D-aspartate (NMDA) receptor, this DAPK1\u2013NR2B interaction increases calcium intake in cells, leading to cell damage. As apoE4 is the strongest genetic risk factor for sporadic Alzheimer\u2019s disease (sAD) and calcium dyshomeostasis is a key driver of sAD-related neurotoxicity, the unclear mechanism linking apoE4 to calcium imbalance remains a critical gap in developing targeted therapies for sAD. Addressing this gap is urgent to advance our understanding of sAD pathogenesis and identify new therapeutic targets. Our study aimed to determine whether the neurotoxic effects of apoE4 are mediated by the interaction between DAPK-1 and the NMDA receptor NR2B subunit.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Primary mouse hippocampal neurons were cultured with apoEs in the presence or absence of an NMDA receptor antagonist that preferentially targets NR2B-containing receptors. Intracellular calcium concentration, cell viability and the proteins related to calcium homeostasis were detected.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Cells treated with apoE4 showed increased levels of intracellular calcium and cell death. The expression of DAPK-1 and NR2B was up-regulated, and interacted with each other under the condition of ApoE4 culture.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>Our study showed that apoE4 causes neurotoxicity by increasing intracellular calcium concentration. The possible mechanism of which was that apoE4 increased the expression of DAPK-1\/NMDAR2B, and further promoted the interaction between the two. These results may provide new targets and ideas for the treatment of AD.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fnins.2026.1845934","type":"journal-article","created":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T06:44:48Z","timestamp":1788417888000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["The role of apolipoprotein E4 on calcium homeostasis in primary hippocampal neurons: an in vitro study"],"prefix":"10.3389","volume":"20","author":[{"given":"Deshan","family":"Liu","sequence":"first","affiliation":[{"name":"Department of Neurology, Fujian Provincial Hospital, Shengli Clinical Medical College of Fujian Medical University, Fuzhou","place":["Fujian, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yifen","family":"Zhuo","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology, Xiamen Haicang Hospital, Xiamen","place":["Fujian, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sisi","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology, Fujian Provincial Hospital, Shengli Clinical Medical College of Fujian Medical University, Fuzhou","place":["Fujian, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yinzhou","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Neurology, Fujian Provincial Hospital, Shengli Clinical Medical College of Fujian Medical University, Fuzhou","place":["Fujian, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaochun","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Neurology, Fujian Medical University Union Hospital, Fuzhou","place":["Fujian, China"]},{"name":"Fujian Institute of Geriatrics, Fujian Medical University Union Hospital, Fuzhou","place":["Fujian, China"]},{"name":"Key Laboratory of Brain Aging and Neurodegenerative Disease, Fujian Key Laboratory of Molecular Neurology, Fujian Medical University, Fuzhou","place":["Fujian, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yusheng","family":"Yao","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology, Fujian Provincial Hospital, Shengli Clinical Medical College of Fujian Medical University, Fuzhou","place":["Fujian, China"]},{"name":"Fujian Provincial Key Laboratory of Critical Care Medicine, Fuzhou","place":["Fujian, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,9,3]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"908","DOI":"10.3390\/brainsci14090908","article-title":"Apolipoprotein E and Alzheimer\u2019s disease in Italian population: Systematic review and meta-analysis.","volume":"14","author":"Abrego-Guandique","year":"2024","journal-title":"Brain Sci."},{"key":"B2","doi-asserted-by":"publisher","first-page":"414","DOI":"10.1016\/j.clnesp.2021.12.030","article-title":"The possible role of ursolic acid in Covid-19: A real game changer.","volume":"47","author":"Al-Kuraishy","year":"2022","journal-title":"Clin. Nutr. ESPEN"},{"key":"B3","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1007\/s12017-025-08857-x","article-title":"Unlocking Alzheimer\u2019s disease: The role of BDNF signaling in neuropathology and treatment.","volume":"27","author":"Alqahtani","year":"2025","journal-title":"Neuromolecular Med."},{"key":"B4","doi-asserted-by":"publisher","first-page":"1385","DOI":"10.4103\/1673-5374.253520","article-title":"Compartmentalized necroptosis activation in excitotoxicity-induced axonal degeneration: A novel mechanism implicated in neurodegenerative disease pathology.","volume":"14","author":"Arrazola","year":"2019","journal-title":"Neural Regen. Res."},{"key":"B5","doi-asserted-by":"publisher","first-page":"10479","DOI":"10.1093\/nar\/gkaa765","article-title":"Post-translational modifications of hnRNP A1 differentially modulate retroviral IRES-mediated translation initiation.","volume":"48","author":"Barrera","year":"2020","journal-title":"Nucleic Acids Res."},{"key":"B6","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1038\/s41422-020-00439-9","article-title":"CaMKII\u03b1-driven, phosphatase-checked postsynaptic plasticity via phase separation.","volume":"31","author":"Cai","year":"2021","journal-title":"Cell Res."},{"key":"B7","doi-asserted-by":"publisher","first-page":"136","DOI":"10.1016\/j.tins.2020.10.004","article-title":"Mitochondria and calcium in Alzheimer\u2019s disease: From cell signaling to neuronal cell death.","volume":"44","author":"Calvo-Rodriguez","year":"2021","journal-title":"Trends Neurosci."},{"key":"B8","doi-asserted-by":"publisher","first-page":"e70038","DOI":"10.1002\/prp2.70038","article-title":"NMDA receptor-mediated Ca2 + flux attenuated by the NMDA receptor\/TRPM4 interface inhibitor brophenexin.","volume":"12","author":"Casby","year":"2024","journal-title":"Pharmacol. Res. Perspect."},{"key":"B9","doi-asserted-by":"publisher","first-page":"2312","DOI":"10.1016\/j.neuron.2023.05.002","article-title":"NMDA receptor functions in health and disease: Old actor, new dimensions.","volume":"111","author":"Dupuis","year":"2023","journal-title":"Neuron"},{"key":"B10","doi-asserted-by":"publisher","first-page":"777","DOI":"10.4103\/1673-5374.322452","article-title":"APOE4 expression is associated with impaired autophagy and mitophagy in astrocytes.","volume":"17","author":"Eran","year":"2022","journal-title":"Neural Regen. Res."},{"key":"B11","doi-asserted-by":"publisher","first-page":"1263","DOI":"10.4103\/1673-5374.327341","article-title":"Mechanosensitivity of N-methyl-D-aspartate receptors (NMDAR) is the key through which amyloid beta oligomers activate them.","volume":"17","author":"Fani","year":"2022","journal-title":"Neural Regen. Res."},{"key":"B12","doi-asserted-by":"publisher","first-page":"2669","DOI":"10.4103\/1673-5374.373714","article-title":"Calcium-permeable AMPA receptors: An underestimated pharmacological target for the therapy of brain pathologies.","volume":"18","author":"Gaidin","year":"2023","journal-title":"Neural Regen. Res."},{"key":"B13","doi-asserted-by":"publisher","first-page":"486","DOI":"10.1038\/s41418-017-0012-4","article-title":"Molecular mechanisms of cell death: Recommendations of the nomenclature committee on cell death 2018.","volume":"25","author":"Galluzzi","year":"2018","journal-title":"Cell Death Differ."},{"key":"B14","doi-asserted-by":"publisher","first-page":"477","DOI":"10.1016\/j.cmet.2024.10.021","article-title":"Mitochondrial calcium uptake declines during aging and is directly activated by oleuropein to boost energy metabolism and skeletal muscle performance.","volume":"37","author":"Gherardi","year":"2025","journal-title":"Cell Metab."},{"key":"B15","doi-asserted-by":"publisher","first-page":"101549","DOI":"10.1016\/j.xpro.2022.101549","article-title":"Immunofluorescence staining of phosphoinositides in primary mouse hippocampal neurons in dissociated culture.","volume":"3","author":"Guo","year":"2022","journal-title":"STAR Protoc."},{"key":"B16","doi-asserted-by":"publisher","first-page":"10103","DOI":"10.1038\/s41467-024-54423-7","article-title":"Functional specificity of liquid-liquid phase separation at the synapse.","volume":"15","author":"Guzikowski","year":"2024","journal-title":"Nat. Commun."},{"key":"B17","doi-asserted-by":"publisher","first-page":"5541","DOI":"10.1113\/jphysiol.2013.258863","article-title":"Comparison of Ca2 + transients and [Ca2 +]i in the dendrites and boutons of non-fast-spiking GABAergic hippocampal interneurons using two-photon laser microscopy and high- and low-affinity dyes.","volume":"591","author":"Kisfali","year":"2013","journal-title":"J. Physiol."},{"key":"B18","doi-asserted-by":"publisher","first-page":"467","DOI":"10.3988\/jcn.2024.0353","article-title":"Impact of apolipoprotein E \u03b54 in Alzheimer\u2019s disease: Insights from a meta-analysis.","volume":"20","author":"Kwon","year":"2024","journal-title":"J. Clin. Neurol."},{"key":"B19","doi-asserted-by":"publisher","first-page":"35","DOI":"10.1186\/s13024-020-00382-8","article-title":"Sex-dependent calcium hyperactivity due to lysosomal-related dysfunction in astrocytes from APOE4 versus APOE3 gene targeted replacement mice.","volume":"15","author":"Larramona-Arcas","year":"2020","journal-title":"Mol. Neurodegener."},{"key":"B20","doi-asserted-by":"publisher","first-page":"382","DOI":"10.4103\/1673-5374.346546","article-title":"Neuroprotective effects of exogenous brain-derived neurotrophic factor on amyloid-beta 1-40-induced retinal degeneration.","volume":"18","author":"Lazaldin","year":"2023","journal-title":"Neural Regen Res."},{"key":"B21","doi-asserted-by":"publisher","first-page":"1075","DOI":"10.1007\/s11427-012-4421-9","article-title":"Zinc reverses glycine-dependent inactivation of NMDARs in cultured rat hippocampal neurons.","volume":"55","author":"Li","year":"2012","journal-title":"Sci. China Life Sci."},{"key":"B22","doi-asserted-by":"publisher","first-page":"1795","DOI":"10.4103\/1673-5374.361532","article-title":"Death-associated protein kinase 1 is associated with cognitive dysfunction in major depressive disorder.","volume":"18","author":"Li","year":"2023","journal-title":"Neural Regen Res."},{"key":"B23","doi-asserted-by":"publisher","first-page":"7","DOI":"10.1186\/s13024-015-0002-2","article-title":"APOE4 enhances age-dependent decline in cognitive function by down-regulating an NMDA receptor pathway in EFAD-Tg mice.","volume":"10","author":"Liu","year":"2015","journal-title":"Mol. Neurodegener."},{"key":"B24","doi-asserted-by":"publisher","first-page":"321","DOI":"10.1016\/j.ijbiomac.2021.01.071","article-title":"CaMK II\/Ca2 + dependent endoplasmic reticulum stress mediates apoptosis of hepatic stellate cells stimulated by transforming growth factor beta 1.","volume":"172","author":"Liu","year":"2021","journal-title":"Int. J. Biol. Macromol."},{"key":"B25","doi-asserted-by":"publisher","first-page":"154594","DOI":"10.1016\/j.phymed.2022.154594","article-title":"Inhibition of GluN2B pathway is involved in the neuroprotective effect of silibinin on streptozotocin-induced Alzheimer\u2019s disease models.","volume":"109","author":"Liu","year":"2023","journal-title":"Phytomedicine"},{"key":"B26","doi-asserted-by":"publisher","first-page":"24646","DOI":"10.1038\/srep24646","article-title":"Effect of the PGD2-DP signaling pathway on primary cultured rat hippocampal neuron injury caused by aluminum overload.","volume":"6","author":"Ma","year":"2016","journal-title":"Sci. Rep."},{"key":"B27","doi-asserted-by":"publisher","first-page":"657","DOI":"10.1016\/j.apsb.2024.10.009","article-title":"Microglial APOE4 promotes neuron degeneration in Alzheimer\u2019s disease through inhibition of lipid droplet autophagy.","volume":"15","author":"Mao","year":"2025","journal-title":"Acta Pharm. Sin. B"},{"key":"B28","doi-asserted-by":"publisher","first-page":"2685","DOI":"10.4103\/1673-5374.335817","article-title":"Implication of 14-3-3\u03b6-BDNF pathway in long-lasting memory enhancement and the rescue from memory deficits.","volume":"17","author":"Masmudi-Mart\u00edn","year":"2022","journal-title":"Neural Regen. Res."},{"key":"B29","doi-asserted-by":"publisher","first-page":"71","DOI":"10.1038\/s41586-020-2247-3","article-title":"APOE4 leads to blood-brain barrier dysfunction predicting cognitive decline.","volume":"581","author":"Montagne","year":"2020","journal-title":"Nature"},{"key":"B30","doi-asserted-by":"publisher","first-page":"2805","DOI":"10.1093\/cvr\/cvab325","article-title":"Genetic inhibition of nuclear factor of activated T-cell c2 prevents atrial fibrillation in CREM transgenic mice.","volume":"118","author":"Ni","year":"2022","journal-title":"Cardiovasc. Res."},{"key":"B31","doi-asserted-by":"publisher","first-page":"eadd6260","DOI":"10.1126\/science.add6260","article-title":"Pharmacological modulation of septins restores calcium homeostasis and is neuroprotective in models of Alzheimer\u2019s disease.","volume":"384","author":"Princen","year":"2024","journal-title":"Science"},{"key":"B32","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1186\/s12974-019-1695-x","article-title":"Dysregulation of BDNF\/TrkB signaling mediated by NMDAR\/Ca2 +\/calpain might contribute to postoperative cognitive dysfunction in aging mice.","volume":"17","author":"Qiu","year":"2020","journal-title":"J. Neuroinflammation"},{"key":"B33","doi-asserted-by":"publisher","first-page":"8686","DOI":"10.1523\/JNEUROSCI.0435-21.2021","article-title":"APOE4 affects basal and NMDAR-mediated protein synthesis in neurons by perturbing calcium homeostasis.","volume":"41","author":"Ramakrishna","year":"2021","journal-title":"J. Neurosci."},{"key":"B34","doi-asserted-by":"publisher","first-page":"238","DOI":"10.3389\/fcimb.2020.00238","article-title":"The PANoptosome: A deadly protein complex driving pyroptosis, apoptosis, and necroptosis (PANoptosis).","volume":"10","author":"Samir","year":"2020","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"B35","doi-asserted-by":"publisher","first-page":"274","DOI":"10.1016\/j.brainresbull.2008.08.002","article-title":"The protective effects of Achyranthes bidentata polypeptides against NMDA-induced cell apoptosis in cultured hippocampal neurons through differential modulation of NR2A- and NR2B-containing NMDA receptors.","volume":"77","author":"Shen","year":"2008","journal-title":"Brain Res. Bull."},{"key":"B36","doi-asserted-by":"publisher","first-page":"2208","DOI":"10.4103\/1673-5374.335793","article-title":"N-methyl-D-aspartate receptor functions altered by neuronal PTP1B activation in Alzheimer\u2019s disease and schizophrenia models.","volume":"17","author":"Stewart","year":"2022","journal-title":"Neural Regen. Res."},{"key":"B37","doi-asserted-by":"publisher","first-page":"222","DOI":"10.1016\/j.cell.2009.12.055","article-title":"DAPK1 interaction with NMDA receptor NR2B subunits mediates brain damage in stroke","volume":"140","author":"Tu","year":"2010","journal-title":"Cell"},{"key":"B38","doi-asserted-by":"publisher","first-page":"379","DOI":"10.1002\/jnr.10138","article-title":"Neurotoxic effects of apolipoprotein E4 are mediated via dysregulation of calcium homeostasis","volume":"67","author":"Veinbergs","year":"2002","journal-title":"J. Neurosci. Res."},{"key":"B39","doi-asserted-by":"publisher","first-page":"513","DOI":"10.1016\/j.jare.2025.01.007","article-title":"Calcium signaling hypothesis: A non-negligible pathogenesis in Alzheimer\u2019s disease.","volume":"77","author":"Wang","year":"2025","journal-title":"J. Adv. Res."},{"key":"B40","doi-asserted-by":"publisher","first-page":"e102161","DOI":"10.1371\/journal.pone.0102161","article-title":"Anti-epileptic effect of Ganoderma lucidum polysaccharides by inhibition of intracellular calcium accumulation and stimulation of expression of CaMKII \u03b1 in epileptic hippocampal neurons.","volume":"9","author":"Wang","year":"2014","journal-title":"PLoS One"},{"key":"B41","doi-asserted-by":"publisher","first-page":"1761","DOI":"10.4103\/1673-5374.331539","article-title":"Do pyroptosis, apoptosis, and necroptosis (PANoptosis) exist in cerebral ischemia? Evidence from cell and rodent studies.","volume":"17","author":"Yan","year":"2022","journal-title":"Neural Regen. Res."},{"key":"B42","doi-asserted-by":"publisher","first-page":"1981","DOI":"10.4103\/NRR.NRR-D-24-00398","article-title":"Acute and chronic excitotoxicity in ischemic stroke and late-onset Alzheimer\u2019s disease.","volume":"20","author":"Yu","year":"2025","journal-title":"Neural Regen. Res."},{"key":"B43","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1038\/s41583-018-0093-1","article-title":"Necroptosis and RIPK1-mediated neuroinflammation in CNS diseases.","volume":"20","author":"Yuan","year":"2019","journal-title":"Nat. Rev. Neurosci."},{"key":"B44","doi-asserted-by":"publisher","first-page":"4","DOI":"10.1186\/s40035-023-00395-5","article-title":"Death-associated protein kinase 1 as a therapeutic target for Alzheimer\u2019s disease.","volume":"13","author":"Zhang","year":"2024","journal-title":"Transl. Neurodegener."},{"key":"B45","doi-asserted-by":"publisher","first-page":"e2410436122","DOI":"10.1073\/pnas.2410436122","article-title":"Ratio-based indicators for cytosolic Ca2 + with visible light excitation.","volume":"122","author":"Zhou","year":"2025","journal-title":"Proc. Natl. Acad. Sci. U. S. A."}],"container-title":["Frontiers in Neuroscience"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fnins.2026.1845934\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T06:44:49Z","timestamp":1788417889000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fnins.2026.1845934\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,3]]},"references-count":45,"alternative-id":["10.3389\/fnins.2026.1845934"],"URL":"https:\/\/doi.org\/10.3389\/fnins.2026.1845934","relation":{},"ISSN":["1662-453X"],"issn-type":[{"value":"1662-453X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,3]]},"article-number":"1845934"}}