{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T11:05:27Z","timestamp":1784804727612,"version":"3.55.0"},"reference-count":52,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2020,10,9]],"date-time":"2020-10-09T00:00:00Z","timestamp":1602201600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100007771","name":"\u015al\u0105ski Uniwersytet Medyczny","doi-asserted-by":"publisher","award":["KNW-2-O41\/D\/9\/N; KNW-1-037\/K\/9\/O"],"award-info":[{"award-number":["KNW-2-O41\/D\/9\/N; KNW-1-037\/K\/9\/O"]}],"id":[{"id":"10.13039\/100007771","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Cobalamin deficiency affects human physiology with sequelae ranging from mild fatigue to severe neuropsychiatric abnormalities. The cellular and molecular aspects of the nervous system disorders associated with hypovitaminosis B12 remain largely unknown. Growing evidence indicates that astrogliosis is an underlying component of a wide range of neuropathologies. Previously, we developed an in vitro model of cobalamin deficiency in normal human astrocytes (NHA) by culturing the cells with c-lactam of hydroxycobalamin (c-lactam OH-Cbl). We revealed a non-apoptotic activation of caspases (3\/7, 8, 9) in cobalamin-deficient NHA, which may suggest astrogliosis. The aim of the current study was to experimentally verify this hypothesis. We indicated an increase in the cellular expression of two astrogliosis markers: glial fibrillary acidic protein and vimentin in cobalamin-deficient NHA using Western blot analysis and immunocytochemistry with confocal laser scanning microscopy. In the next step of the study, we revealed c-lactam OH-Cbl as a potential non-toxic vitamin B12 antagonist in an in vivo model using zebrafish embryos. We believe that the presented results will contribute to a better understanding of the cellular mechanism underlying neurologic pathology due to cobalamin deficiency and will serve as a foundation for further studies.<\/jats:p>","DOI":"10.3390\/cells9102261","type":"journal-article","created":{"date-parts":[[2020,10,15]],"date-time":"2020-10-15T21:42:21Z","timestamp":1602798141000},"page":"2261","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Astrogliosis in an Experimental Model of Hypovitaminosis B12: A Cellular Basis of Neurological Disorders due to Cobalamin Deficiency"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7892-0462","authenticated-orcid":false,"given":"Zuzanna","family":"Rzepka","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences in Sosnowiec, Medical University of Silesia in Katowice, Jagiello\u0144ska 4, 41-200 Sosnowiec, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8260-4339","authenticated-orcid":false,"given":"Jakub","family":"Rok","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences in Sosnowiec, Medical University of Silesia in Katowice, Jagiello\u0144ska 4, 41-200 Sosnowiec, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Justyna","family":"Kowalska","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences in Sosnowiec, Medical University of Silesia in Katowice, Jagiello\u0144ska 4, 41-200 Sosnowiec, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Klaudia","family":"Banach","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences in Sosnowiec, Medical University of Silesia in Katowice, Jagiello\u0144ska 4, 41-200 Sosnowiec, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3985-4244","authenticated-orcid":false,"given":"Justyna Magdalena","family":"Hermanowicz","sequence":"additional","affiliation":[{"name":"Department of Pharmacodynamics, Medical University of Bialystok, Mickiewicza 2C, 15-222 Bialystok, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2247-3004","authenticated-orcid":false,"given":"Artur","family":"Beberok","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences in Sosnowiec, Medical University of Silesia in Katowice, Jagiello\u0144ska 4, 41-200 Sosnowiec, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8384-7116","authenticated-orcid":false,"given":"Beata","family":"Sieklucka","sequence":"additional","affiliation":[{"name":"Department of Pharmacodynamics, Medical University of Bialystok, Mickiewicza 2C, 15-222 Bialystok, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5197-4222","authenticated-orcid":false,"given":"Dorota","family":"Gryko","sequence":"additional","affiliation":[{"name":"Institute of Organic Chemistry, Polish Academy of Science, Kasprzaka 44\/52, 01-224 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6918-3330","authenticated-orcid":false,"given":"Dorota","family":"Wrze\u015bniok","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences in Sosnowiec, Medical University of Silesia in Katowice, Jagiello\u0144ska 4, 41-200 Sosnowiec, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,10,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"17040","DOI":"10.1038\/nrdp.2017.40","article-title":"Vitamin B12 deficiency","volume":"3","author":"Green","year":"2017","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1016\/j.mpmed.2017.01.007","article-title":"Iron, vitamin B12 and folate","volume":"45","author":"Moll","year":"2017","journal-title":"Medicine"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1016\/j.mayocpiqo.2019.03.002","article-title":"The many faces of cobalamin (vitamin B12) deficiency","volume":"3","author":"Wolffenbuttel","year":"2019","journal-title":"Mayo Clin. Proc. Innov. Qual. Outcomes"},{"key":"ref_4","first-page":"809","article-title":"Effect of hydroxycobalamin[c-lactam] on propionate and carnitine metabolism in the rat","volume":"266","author":"Brass","year":"1990","journal-title":"Biochem. J."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1422","DOI":"10.1172\/JCI115148","article-title":"Inhibition of cobalamin-dependent enzymes by cobalamin analogues in rats","volume":"87","author":"Stabler","year":"1991","journal-title":"J. Clin. Investig."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1016\/S0304-3940(00)01243-X","article-title":"Inhibition of vitamin B12 metabolism by OH-cobalamin c-lactam in rat oligodendrocytes in culture: A model for studying neuropathy due to vitamin B12 deficiency","volume":"288","author":"Sponne","year":"2000","journal-title":"Neurosci. Lett."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"720","DOI":"10.1007\/s10545-009-1197-6","article-title":"Long-term exposure of human proximal tubule cells to hydroxycobalamin[c-lactam] as a possible model to study renal disease in methylmalonic acidurias","volume":"32","author":"Sauer","year":"2009","journal-title":"J. Inherit. Metab. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1016\/j.tox.2015.07.015","article-title":"Impaired mitochondrial function in HepG2 cells treated with hydroxy-cobalamin[c-lactam]: A cell model for idiosyncratic toxicity","volume":"336","author":"Haegler","year":"2015","journal-title":"Toxicology"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Haegler, P., Gr\u00fcnig, D., Berger, B., Terracciano, L., Kr\u00e4henb\u00fchl, S., and Bouitbir, J. (2017). Hepatic effects of pharmacological doses of hydroxy-cobalamin[c-lactam] in mice. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0171026"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1264","DOI":"10.1002\/cbic.201500072","article-title":"Antivitamins for medicinal applications","volume":"16","author":"Zelder","year":"2015","journal-title":"Chembiochem"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Rzepka, Z., Respondek, M., Pawlik, J., Beberok, A., Gryko, D., and Wrze\u015bniok, D. (2019). Cobalamin deficiency: Effect on homeostasis of cultured human astrocytes. Cells, 8.","DOI":"10.3390\/cells8121505"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/j.brainres.2013.03.033","article-title":"High-fat diet feeding causes rapid, non-apoptotic cleavage of caspase-3 in astrocytes","volume":"1512","author":"Guyenet","year":"2013","journal-title":"Brain Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1016\/j.brainres.2012.02.056","article-title":"Caspase activation contributes to astrogliosis","volume":"1450","author":"Aras","year":"2012","journal-title":"Brain Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1016\/j.neuroscience.2008.01.054","article-title":"Survivin and heat shock protein 25\/27 colocalize with cleaved caspase-3 in surviving reactive astrocytes following excitotoxicity to the immature brain","volume":"153","author":"Villapol","year":"2008","journal-title":"Neuroscience"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1016\/j.pneurobio.2016.01.001","article-title":"Astrogliosis: An integral player in the pathogenesis of Alzheimer\u2019s disease","volume":"144","author":"Osborn","year":"2016","journal-title":"Prog. Neurobiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2739","DOI":"10.1007\/s00018-019-03111-7","article-title":"Astrocyte alterations in neurodegenerative pathologies and their modeling in human induced pluripotent stem cell platforms","volume":"76","author":"Oksanen","year":"2019","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1007\/s12035-010-8098-4","article-title":"Astrogliosis in CNS pathologies: Is there a role for microglia?","volume":"41","author":"Zhang","year":"2010","journal-title":"Mol. Neurobiol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1038\/cdd.2008.172","article-title":"Astroglia in dementia and Alzheimer\u2019s disease","volume":"16","author":"Olabarria","year":"2009","journal-title":"Cell Death Differ."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Rzepka, Z., Respondek, M., Beberok, A., \u00d3 Proinsias, K., Gryko, D., and Wrze\u015bniok, D. (2018). Vitamin B12 deficiency induces imbalance in melanocytes homeostasis\u2014A cellular basis of hypocobalaminemia pigmentary manifestations. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19092845"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"496","DOI":"10.1016\/j.yrtph.2014.05.018","article-title":"OECD validation study to assess intra- and inter-laboratory reproducibility of the zebrafish embryo toxicity test for acute aquatic toxicity testing","volume":"69","author":"Busquet","year":"2014","journal-title":"Regul. Toxicol. Pharmacol."},{"key":"ref_21","unstructured":"OECD (2013). Test No. 236: Fish Embryo Acute Toxicity (FET) Test, OECD Publishing."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1007\/s11062-016-9568-8","article-title":"Glial fibrillary acidic protein (GFAP): On the 45th anniversary of its discovery","volume":"48","author":"Tykhomyrov","year":"2016","journal-title":"Neurophysiology"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1007\/978-1-61779-452-0_21","article-title":"An in vitro model of reactive astrogliosis and its effect on neuronal growth","volume":"814","author":"Yu","year":"2012","journal-title":"Methods Mol Biol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Danielsson, F., Peterson, M.K., Caldeira Ara\u00fajo, H., Lautenschl\u00e4ger, F., and Gad, A.K. (2018). Vimentin diversity in health and disease. Cells, 7.","DOI":"10.3390\/cells7100147"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Lopez-Rodriguez, A.B., Acaz-Fonseca, E., Viveros, M.P., and Garcia-Segura, L.M. (2015). Changes in cannabinoid receptors, aquaporin 4 and vimentin expression after traumatic brain injury in adolescent male mice. Association with edema and neurological deficit. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0128782"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1077","DOI":"10.1152\/physrev.00041.2013","article-title":"Astrocyte reactivity and reactive astrogliosis: Costs and benefits","volume":"94","author":"Pekny","year":"2014","journal-title":"Physiol. Rev."},{"key":"ref_27","first-page":"311","article-title":"Vimentin and GFAP responses in astrocytes after contusion trauma to the murine brain","volume":"28","author":"Israelsson","year":"2010","journal-title":"Restor. Neurol. Neurosci."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"6391","DOI":"10.1523\/JNEUROSCI.6221-11.2012","article-title":"Genomic analysis of reactive astrogliosis","volume":"32","author":"Zamanian","year":"2012","journal-title":"J. Neurosci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1007\/s00418-013-1110-0","article-title":"Intermediate filaments are important for astrocyte response to oxidative stress induced by oxygen-glucose deprivation and reperfusion","volume":"140","author":"Nilsson","year":"2013","journal-title":"Histochem. Cell Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"5016","DOI":"10.1523\/JNEUROSCI.0820-04.2004","article-title":"Absence of glial fibrillary acidic protein and vimentin prevents hypertrophy of astrocytic processes and improves post-traumatic regeneration","volume":"24","author":"Wilhelmsson","year":"2004","journal-title":"J. Neurosci."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Pablo, Y.D., Marasek, P., Pozo-Rodrig\u00e1lvarez, A., Wilhelmsson, U., Inagaki, M., Pekna, M., and Pekny, M. (2019). Vimentin phosphorylation is required for normal cell division of immature astrocytes. Cells, 8.","DOI":"10.3390\/cells8091016"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1503","DOI":"10.1038\/s41418-018-0058-y","article-title":"RGMa mediates reactive astrogliosis and glial scar formation through TGF\u03b21\/Smad2\/3 signaling after stroke","volume":"25","author":"Zhang","year":"2018","journal-title":"Cell Death Differ."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"474","DOI":"10.1111\/nan.12420","article-title":"Astrogliosis and impaired aquaporin-4 and dystrophin systems in idiopathic normal pressure hydrocephalus","volume":"44","author":"Eide","year":"2018","journal-title":"Neuropathol. Appl. Neurobiol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"166","DOI":"10.1007\/s11064-017-2385-7","article-title":"SOD1 mutations causing familial amyotrophic lateral sclerosis induce toxicity in astrocytes: Evidence for bystander effects in a continuum of astrogliosis","volume":"43","author":"Wallis","year":"2018","journal-title":"Neurochem. Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1921","DOI":"10.1007\/s00210-020-01905-2","article-title":"Agathisflavone modulates astrocytic responses and increases the population of neurons in an in vitro model of traumatic brain injury","volume":"393","author":"David","year":"2020","journal-title":"Naunyn Schmiedebergs Arch. Pharmacol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"204","DOI":"10.3109\/00207454.2016.1173692","article-title":"Astrocytes: Inexplicable cells in neurodegeneration","volume":"127","author":"Singh","year":"2017","journal-title":"Int. J. Neurosci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1186\/s40478-020-00919-4","article-title":"Abnormal mitosis in reactive astrocytes","volume":"8","author":"Sosunov","year":"2020","journal-title":"Acta Neuropathol. Commun."},{"key":"ref_38","first-page":"1318","article-title":"Dodecylamine derivative of hydroxocobalamin acts as a potent inhibitor of cobalamin-dependent methionine synthase in mammalian cultured COS-7 cells","volume":"5","author":"Bito","year":"2014","journal-title":"Food Nutr. Sci."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Wasel, O., and Freeman, J.L. (2020). Chemical and genetic zebrafish models to define mechanisms of and treatments for dopaminergic neurodegeneration. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21175981"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1002\/glia.23083","article-title":"Identification of a conserved and acute neurodegeneration-specific microglial transcriptome in the zebrafish","volume":"65","author":"Oosterhof","year":"2017","journal-title":"Glia"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Koiwa, J., Shiromizu, T., Adachi, Y., Ikejiri, M., Nakatani, K., Tanaka, T., and Nishimura, Y. (2019). Generation of a triple-transgenic zebrafish line for assessment of developmental neurotoxicity during neuronal differentiation. Pharmaceuticals (Basel), 12.","DOI":"10.3390\/ph12040145"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1038\/s41420-018-0109-7","article-title":"Zebrafish: An emerging real-time model system to study Alzheimer\u2019s disease and neurospecific drug discovery","volume":"4","author":"Saleem","year":"2018","journal-title":"Cell. Death. Discov."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1297","DOI":"10.1038\/s41593-020-0703-x","article-title":"Live-imaging of astrocyte morphogenesis and function in zebrafish neural circuits","volume":"23","author":"Chen","year":"2020","journal-title":"Nat. Neurosci."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"121639","DOI":"10.1016\/j.jhazmat.2019.121639","article-title":"Neurotoxic effects of aflatoxin B1 on human astrocytes in vitro and on glial cell development in zebrafish in vivo","volume":"386","author":"Park","year":"2020","journal-title":"J. Hazard. Mater."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"364","DOI":"10.1016\/j.ecoenv.2018.06.009","article-title":"Billings reservoir water used for human consumption presents microbiological contaminants and induces both behavior impairments and astrogliosis in zebrafish","volume":"161","author":"Leme","year":"2018","journal-title":"Ecotoxicol. Environ. Saf."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1016\/j.expneurol.2018.04.008","article-title":"Glia maturation factor beta is required for reactive gliosis after traumatic brain injury in zebrafish","volume":"305","author":"Yin","year":"2018","journal-title":"Exp. Neurol."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Greibe, E., Fedosov, S., and Nexo, E. (2012). The cobalamin-binding protein in zebrafish is an intermediate between the three cobalamin-binding proteins in human. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0035660"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"17606","DOI":"10.1074\/jbc.RA118.005323","article-title":"Functional and phylogenetic characterization of noncanonical vitamin B12-binding proteins in zebrafish suggests involvement in cobalamin transport","volume":"293","author":"Benoit","year":"2018","journal-title":"J. Biol. Chem."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"282","DOI":"10.1111\/j.1346-8138.2001.tb00134.x","article-title":"Generalized hyperpigmentation of the skin due to vitamin B12 deficiency","volume":"28","author":"Mori","year":"2001","journal-title":"J. Dermatol."},{"key":"ref_50","first-page":"551","article-title":"Reversible cutaneous hyperpigmentation and nails with white hair due to vitamin B12 deficiency","volume":"17","author":"Niiyama","year":"2007","journal-title":"Eur. J. Dermatol."},{"key":"ref_51","first-page":"529","article-title":"Cutaneous lesions and vitamin B12 deficiency: An often-forgotten link","volume":"54","author":"Kannan","year":"2008","journal-title":"Can. Fam. Physician"},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Lee, M.S., Bonner, J.R., Bernard, D.J., Sanchez, E.L., Sause, E.T., Prentice, R.R., Burgess, S.M., and Brody, L.C. (2012). Disruption of the folate pathway in zebrafish causes developmental defects. BMC Dev. Biol., 12.","DOI":"10.1186\/1471-213X-12-12"}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/9\/10\/2261\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:17:49Z","timestamp":1760177869000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/9\/10\/2261"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,10,9]]},"references-count":52,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2020,10]]}},"alternative-id":["cells9102261"],"URL":"https:\/\/doi.org\/10.3390\/cells9102261","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,10,9]]}}}