{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T05:44:56Z","timestamp":1790142296603,"version":"4.0.1"},"reference-count":0,"publisher":"Georg Thieme Verlag KG","issue":"04","license":[{"start":{"date-parts":[[2020,4,14]],"date-time":"2020-04-14T00:00:00Z","timestamp":1586822400000},"content-version":"vor","delay-in-days":13,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"name":"Auckland Medical Research Foundation, Leukemia & Blood Cancer New Zealand and Cancer Research Trust"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Thromb Haemost"],"published-print":{"date-parts":[[2020,4]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The release of calcium ions (Ca2+) from the endoplasmic reticulum (ER) and related store-operated calcium entry (SOCE) regulate maturation of normal megakaryocytes. The N-methyl-D-aspartate (NMDA) receptor (NMDAR) provides an additional mechanism for Ca2+ influx in megakaryocytic cells, but its role remains unclear. We created a model of NMDAR hypofunction in Meg-01 cells using CRISPR-Cas9 mediated knockout of the GRIN1 gene, which encodes an obligate, GluN1 subunit of the NMDAR. We found that compared with unmodified Meg-01 cells, Meg-01-GRIN1\n          \u2212\/\u2212 cells underwent atypical differentiation biased toward erythropoiesis, associated with increased basal ER stress and cell death. Resting cytoplasmic Ca2+ levels were higher in Meg-01-GRIN1\n          \u2212\/\u2212 cells, but ER Ca2+ release and SOCE were lower after activation. Lysosome-related organelles accumulated including immature dense granules that may have contributed an alternative source of intracellular Ca2+. Microarray analysis revealed that Meg-01-GRIN1\n          \u2212\/\u2212 cells had deregulated expression of transcripts involved in Ca2+ metabolism, together with a shift in the pattern of hematopoietic transcription factors toward erythropoiesis. In keeping with the observed pro-cell death phenotype induced by GRIN1 deletion, memantine (NMDAR inhibitor) increased cytotoxic effects of cytarabine in unmodified Meg-01 cells. In conclusion, NMDARs comprise an integral component of the Ca2+ regulatory network in Meg-01 cells that help balance ER stress and megakaryocytic-erythroid differentiation. We also provide the first evidence that megakaryocytic NMDARs regulate biogenesis of lysosome-related organelles, including dense granules. Our results argue that intracellular Ca2+ homeostasis may be more important for normal megakaryocytic and erythroid differentiation than currently recognized; thus, modulation may offer therapeutic opportunities.<\/jats:p>","DOI":"10.1055\/s-0040-1708483","type":"journal-article","created":{"date-parts":[[2020,4,14]],"date-time":"2020-04-14T22:31:23Z","timestamp":1586903483000},"page":"671-686","source":"Crossref","is-referenced-by-count":16,"title":["N-Methyl-D-Aspartate Receptor Hypofunction in Meg-01 Cells Reveals a Role for Intracellular Calcium Homeostasis in Balancing Megakaryocytic-Erythroid Differentiation"],"prefix":"10.1055","volume":"120","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3988-2659","authenticated-orcid":false,"given":"James I.","family":"Hearn","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine and Pathology, School of Medical Sciences, University of Auckland, Auckland, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Taryn N.","family":"Green","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine and Pathology, School of Medical Sciences, University of Auckland, Auckland, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin","family":"Chopra","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine and Pathology, School of Medical Sciences, University of Auckland, Auckland, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yohanes N. S.","family":"Nursalim","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine and Pathology, School of Medical Sciences, University of Auckland, Auckland, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leandro","family":"Ladvanszky","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine and Pathology, School of Medical Sciences, University of Auckland, Auckland, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicholas","family":"Knowlton","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine and Pathology, School of Medical Sciences, University of Auckland, Auckland, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cherie","family":"Blenkiron","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine and Pathology, School of Medical Sciences, University of Auckland, Auckland, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raewyn C.","family":"Poulsen","sequence":"additional","affiliation":[{"name":"Department of Medicine, School of Medicine, University of Auckland, Auckland, New Zealand"},{"name":"Department of Pharmacology and Clinical Pharmacology, School of Medical Sciences, University of Auckland, Auckland, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dean C.","family":"Singleton","sequence":"additional","affiliation":[{"name":"Auckland Cancer Society Research Centre, University of Auckland, Auckland, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan K.","family":"Bohlander","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine and Pathology, School of Medical Sciences, University of Auckland, Auckland, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8378-8048","authenticated-orcid":false,"given":"Maggie L.","family":"Kalev-Zylinska","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine and Pathology, School of Medical Sciences, University of Auckland, Auckland, New Zealand"},{"name":"LabPlus Haematology, Auckland City Hospital, Auckland, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"194","published-online":{"date-parts":[[2020,4,14]]},"container-title":["Thrombosis and Haemostasis"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/www.thieme-connect.de\/products\/ejournals\/pdf\/10.1055\/s-0040-1708483.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,4,14]],"date-time":"2020-04-14T22:31:57Z","timestamp":1586903517000},"score":1,"resource":{"primary":{"URL":"http:\/\/www.thieme-connect.de\/DOI\/DOI?10.1055\/s-0040-1708483"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,4]]},"references-count":0,"journal-issue":{"issue":"04","published-online":{"date-parts":[[2020,4,14]]},"published-print":{"date-parts":[[2020,4]]}},"URL":"https:\/\/doi.org\/10.1055\/s-0040-1708483","relation":{},"ISSN":["0340-6245","2567-689X"],"issn-type":[{"value":"0340-6245","type":"print"},{"value":"2567-689X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,4]]}}}