{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T12:03:24Z","timestamp":1790424204461,"version":"4.1.0"},"reference-count":58,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2021,5,8]],"date-time":"2021-05-08T00:00:00Z","timestamp":1620432000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100010663","name":"H2020 European Research Council","doi-asserted-by":"publisher","award":["Grant-2019-WATCH-810331"],"award-info":[{"award-number":["Grant-2019-WATCH-810331"]}],"id":[{"id":"10.13039\/100010663","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100004807","name":"DFG","doi-asserted-by":"publisher","award":["GRK1957"],"award-info":[{"award-number":["GRK1957"]}],"id":[{"id":"10.13039\/100004807","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The mechanisms underlying the transport of leptin into the brain are still largely unclear. While the leptin receptor has been implicated in the transport process, recent evidence has suggested an additional role of LRP2 (megalin). To evaluate the function of LRP2 for leptin transport across the blood-brain barrier (BBB), we developed a novel leptin-luciferase fusion protein (pLG), which stimulated leptin signaling and was transported in an in vitro BBB model based on porcine endothelial cells. The LRP inhibitor RAP did not affect leptin transport, arguing against a role of LRP2. In line with this, the selective deletion of LRP2 in brain endothelial cells and epithelial cells of the choroid plexus did not influence bodyweight, body composition, food intake, or energy expenditure of mice. These findings suggest that LRP2 at the BBB is not involved in the transport of leptin into the brain, nor in the development of obesity as has previously been described.<\/jats:p>","DOI":"10.3390\/ijms22094998","type":"journal-article","created":{"date-parts":[[2021,5,9]],"date-time":"2021-05-09T21:32:46Z","timestamp":1620595966000},"page":"4998","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Is LRP2 Involved in Leptin Transport over the Blood-Brain Barrier and Development of Obesity?"],"prefix":"10.3390","volume":"22","author":[{"given":"Elvira S.","family":"Sandin","sequence":"first","affiliation":[{"name":"Institute for Experimental and Clinical Pharmacology and Toxicology, Center of Brain, Behavior and Metabolism, University of L\u00fcbeck, 23562 L\u00fcbeck, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julica","family":"Folberth","sequence":"additional","affiliation":[{"name":"Institute for Experimental and Clinical Pharmacology and Toxicology, Center of Brain, Behavior and Metabolism, University of L\u00fcbeck, 23562 L\u00fcbeck, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2815-4426","authenticated-orcid":false,"given":"Helge","family":"M\u00fcller-Fielitz","sequence":"additional","affiliation":[{"name":"Institute for Experimental and Clinical Pharmacology and Toxicology, Center of Brain, Behavior and Metabolism, University of L\u00fcbeck, 23562 L\u00fcbeck, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claus U.","family":"Pietrzik","sequence":"additional","affiliation":[{"name":"Institute for Pathobiochemistry, University Medical Center of Johannes Gutenberg University Mainz, 55099 Mainz, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elisabeth","family":"Herold","sequence":"additional","affiliation":[{"name":"Institute of Biochemistry, Center of Structural and Cell Biology in Medicine, University of L\u00fcbeck, 23562 L\u00fcbeck, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas E.","family":"Willnow","sequence":"additional","affiliation":[{"name":"Molecular Cardiovascular Research, Max-Delbrueck-Center for Molecular Medicine, 13092 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul T.","family":"Pfluger","sequence":"additional","affiliation":[{"name":"Research Unit Neurobiology of Diabetes, Helmholtz Zentrum M\u00fcnchen and Technical University Munich, 85764 Neuherberg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9976-9930","authenticated-orcid":false,"given":"Ruben","family":"Nogueiras","sequence":"additional","affiliation":[{"name":"CIMUS, Universidade de Santiago de Compostela-Instituto de Investigaci\u00f3n Sanitaria, 15782 Santiago de Compostela, Spain"},{"name":"CIBER Fisiopatolog\u00eda de la Obesidad y Nutrici\u00f3n (CIBERobn), 15782 Santiago de Compostela, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7185-3615","authenticated-orcid":false,"given":"Vincent","family":"Prevot","sequence":"additional","affiliation":[{"name":"Lille Neuroscience &amp; Cognition, Laboratory of Development and Plasticity of the Neuroendocrine Brain, Inserm, CHU Lille, University Lille, UMR-S1172, EGID, DISTALZ, F-59000 Lille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4548-7241","authenticated-orcid":false,"given":"Thomas","family":"Krey","sequence":"additional","affiliation":[{"name":"Institute of Biochemistry, Center of Structural and Cell Biology in Medicine, University of L\u00fcbeck, 23562 L\u00fcbeck, Germany"},{"name":"German Center for Infection Research (DZIF), Partner Site Hamburg-L\u00fcbeck-Borstel-Riems, 22607 Hamburg, Germany"},{"name":"Excellence Cluster 2155 RESIST, Hannover Medical School, 30625 Hannover, Germany"},{"name":"Centre for Structural Systems Biology (CSSB), 22607 Hamburg, Germany"},{"name":"Institute of Virology, Hannover Medical School, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4510-9718","authenticated-orcid":false,"given":"Markus","family":"Schwaninger","sequence":"additional","affiliation":[{"name":"Institute for Experimental and Clinical Pharmacology and Toxicology, Center of Brain, Behavior and Metabolism, University of L\u00fcbeck, 23562 L\u00fcbeck, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,5,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"754","DOI":"10.1038\/s42255-019-0095-y","article-title":"Leptin and the endocrine control of energy balance","volume":"1","author":"Friedman","year":"2019","journal-title":"Nat. Metab."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"543","DOI":"10.1126\/science.7624777","article-title":"Weight-reducing effects of the plasma protein encoded by the obese gene","volume":"269","author":"Halaas","year":"1995","journal-title":"Science"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.molmet.2017.12.001","article-title":"The LepR-mediated leptin transport across brain barriers controls food reward","volume":"8","author":"Sandin","year":"2018","journal-title":"Mol. Metab."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1016\/j.tips.2013.11.003","article-title":"Feelings about food: The ventral tegmental area in food reward and emotional eating","volume":"35","author":"Meye","year":"2014","journal-title":"Trends Pharmacol. Sci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"741","DOI":"10.1016\/j.cmet.2015.08.003","article-title":"Leptin Suppresses the Rewarding Effects of Running via STAT3 Signaling in Dopamine Neurons","volume":"22","author":"Fernandes","year":"2015","journal-title":"Cell Metab."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"856","DOI":"10.1038\/nature01388","article-title":"STAT3 signalling is required for leptin regulation of energy balance but not reproduction","volume":"421","author":"Bates","year":"2003","journal-title":"Nature"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1038\/ng0996-95","article-title":"Leptin activation of Stat3 in the hypothalamus of wild-type and ob\/ob mice but not db\/db mice","volume":"14","author":"Vaisse","year":"1996","journal-title":"Nat. Genet."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1038\/nrn3745","article-title":"Neurobiology of food intake in health and disease","volume":"15","author":"Morton","year":"2014","journal-title":"Nat. Rev. Neurosci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1016\/j.neuron.2016.08.032","article-title":"Hunger-Driven Motivational State Competition","volume":"92","author":"Burnett","year":"2016","journal-title":"Neuron"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"763","DOI":"10.1038\/s42255-020-0229-2","article-title":"Profound and redundant functions of arcuate neurons in obesity development","volume":"2","author":"Zhu","year":"2020","journal-title":"Nat. Metab."},{"key":"ref_11","unstructured":"(2020). World Health Statistics 2020: Monitoring Health for the SDGs, Sustainable Development Goals, World Health Organization."},{"key":"ref_12","unstructured":"(2020). 2020 Global Nutrition Report: Food Systems and Nutrition Equity, The Global Nutrition Report\u2019s Independent Expert Group."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1311","DOI":"10.1038\/nm1295-1311","article-title":"Leptin levels reflect body lipid content in mice: Evidence for diet-induced resistance to leptin action","volume":"1","author":"Frederich","year":"1995","journal-title":"Nat. Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1016\/j.cmet.2012.07.004","article-title":"Selective insulin and leptin resistance in metabolic disorders","volume":"16","year":"2012","journal-title":"Cell Metab."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1172\/JCI119171","article-title":"Diet-induced obese mice develop peripheral, but not central, resistance to leptin","volume":"99","author":"Compton","year":"1997","journal-title":"J. Clin. Investig."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"8878","DOI":"10.1073\/pnas.94.16.8878","article-title":"Physiological response to long-term peripheral and central leptin infusion in lean and obese mice","volume":"94","author":"Halaas","year":"1997","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"444","DOI":"10.1038\/s41574-019-0213-7","article-title":"The blood-brain barrier as an endocrine tissue","volume":"15","author":"Banks","year":"2019","journal-title":"Nat. Rev. Endocrinol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1016\/j.molmet.2013.09.001","article-title":"Hunting for the functions of short leptin receptor isoforms","volume":"2","author":"Dam","year":"2013","journal-title":"Mol. Metab."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1449","DOI":"10.1016\/S0196-9781(99)00156-4","article-title":"Decreased transport of leptin across the blood-brain barrier in rats lacking the short form of the leptin receptor","volume":"20","author":"Kastin","year":"1999","journal-title":"Peptides"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"775","DOI":"10.1210\/endo.143.3.8669","article-title":"Characterizaton of short isoforms of the leptin receptor in rat cerebral microvessels and of brain uptake of leptin in mouse models of obesity","volume":"143","author":"Hileman","year":"2002","journal-title":"Endocrinology"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2281","DOI":"10.1096\/fj.09-143487","article-title":"Unique leptin trafficking by a tailless receptor","volume":"24","author":"Tu","year":"2010","journal-title":"FASEB J."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1392","DOI":"10.1016\/j.peptides.2011.05.011","article-title":"Effects of cell-type specific leptin receptor mutation on leptin transport across the BBB","volume":"32","author":"Hsuchou","year":"2011","journal-title":"Peptides"},{"key":"ref_23","first-page":"1","article-title":"Inhibition of Leptin-ObR Interaction Does not Prevent Leptin Translocation Across a Human Blood-Brain Barrier Model","volume":"28","author":"Goode","year":"2016","journal-title":"J. Neuroendocrinol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1016\/j.molmet.2019.12.013","article-title":"Identification of the leptin receptor sequences crucial for the STAT3-Independent control of metabolism","volume":"32","author":"Barnes","year":"2020","journal-title":"Mol. Metab."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"5339","DOI":"10.2741\/3599","article-title":"Functional roles of short sequence motifs in the endocytosis of membrane receptors","volume":"14","author":"Pandey","year":"2009","journal-title":"Front. Biosci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1006\/exnr.1999.7052","article-title":"Glycoprotein 330\/megalin (LRP-2) has low prevalence as mRNA and protein in brain microvessels and choroid plexus","volume":"157","author":"Chun","year":"1999","journal-title":"Exp. Neurol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"4229","DOI":"10.1073\/pnas.93.9.4229","article-title":"Glycoprotein 330\/megalin: Probable role in receptor-mediated transport of apolipoprotein J alone and in a complex with Alzheimer disease amyloid beta at the blood-brain and blood-cerebrospinal fluid barriers","volume":"93","author":"Zlokovic","year":"1996","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1016\/j.cmet.2013.12.015","article-title":"Hypothalamic tanycytes are an ERK-gated conduit for leptin into the brain","volume":"19","author":"Balland","year":"2014","journal-title":"Cell Metab."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"258","DOI":"10.1038\/nrm778","article-title":"Megalin and cubilin: Multifunctional endocytic receptors","volume":"3","author":"Christensen","year":"2002","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"902","DOI":"10.1016\/j.neurobiolaging.2007.01.008","article-title":"Megalin mediates the transport of leptin across the blood-CSF barrier","volume":"29","author":"Dietrich","year":"2008","journal-title":"Neurobiol. Aging"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"10884","DOI":"10.1523\/JNEUROSCI.2909-05.2005","article-title":"Choroid plexus megalin is involved in neuroprotection by serum insulin-like growth factor I","volume":"25","author":"Carro","year":"2005","journal-title":"J. Neurosci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"3935","DOI":"10.1210\/en.2004-0074","article-title":"Evidence indicating that renal tubular metabolism of leptin is mediated by megalin but not by the leptin receptors","volume":"145","author":"Hama","year":"2004","journal-title":"Endocrinology"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1186\/s12974-017-0800-2","article-title":"Obesity and neuroinflammatory phenotype in mice lacking endothelial megalin","volume":"14","author":"Bartolome","year":"2017","journal-title":"J. Neuroinflamm."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"230","DOI":"10.1006\/dbio.2000.0106","article-title":"Tie2-Cre transgenic mice: A new model for endothelial cell-lineage analysis in vivo","volume":"230","author":"Kisanuki","year":"2001","journal-title":"Dev. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1016\/j.bbadis.2015.10.006","article-title":"Genetic manipulation of brain endothelial cells in vivo","volume":"1862","author":"Assmann","year":"2016","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2615","DOI":"10.1084\/jem.20110398","article-title":"TAK1 in brain endothelial cells mediates fever and lethargy","volume":"208","author":"Ridder","year":"2011","journal-title":"J. Exp. Med."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"862","DOI":"10.1177\/0271678X16630991","article-title":"In vitro models of the blood-brain barrier: An overview of commonly used brain endothelial cell culture models and guidelines for their use","volume":"36","author":"Helms","year":"2016","journal-title":"J. Cereb. Blood Flow Metab."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1016\/0304-4157(94)90005-1","article-title":"The alpha 2-macroglobulin receptor and epithelial glycoprotein-330: Two giant receptors mediating endocytosis of multiple ligands","volume":"1197","author":"Moestrup","year":"1994","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"897","DOI":"10.1083\/jcb.201504057","article-title":"TDP-43 is intercellularly transmitted across axon terminals","volume":"211","author":"Feiler","year":"2015","journal-title":"J. Cell Biol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1083\/jcb.201609035","article-title":"A targeted RNAi screen identifies factors affecting diverse stages of receptor-mediated transcytosis","volume":"216","author":"Nelms","year":"2017","journal-title":"J. Cell Biol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1096\/fj.02-0578fje","article-title":"Hypocalcemia and osteopathy in mice with kidney-specific megalin gene defect","volume":"17","author":"Leheste","year":"2003","journal-title":"FASEB J."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1186\/s12974-020-1702-2","article-title":"Endothelial-specific deficiency of megalin in the brain protects mice against high-fat diet challenge","volume":"17","author":"Bartolome","year":"2020","journal-title":"J. Neuroinflamm."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1002\/ar.a.10123","article-title":"Differential distribution of cubilin and megalin expression in the mouse embryo","volume":"277","author":"Drake","year":"2004","journal-title":"Anat. Rec. A Discov. Mol. Cell Evol. Biol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1928","DOI":"10.1182\/blood-2016-02-697870","article-title":"Identification of novel regulators of developmental hematopoiesis using Endoglin regulatory elements as molecular probes","volume":"128","author":"Nasrallah","year":"2016","journal-title":"Blood"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"4261","DOI":"10.1172\/JCI128164","article-title":"Lactoferrin-induced myeloid-derived suppressor cell therapy attenuates pathologic inflammatory conditions in newborn mice","volume":"129","author":"Liu","year":"2019","journal-title":"J. Clin. Investig."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"711","DOI":"10.3233\/JAD-131604","article-title":"Alzheimer\u2019s disease-like impaired cognition in endothelial-specific megalin-null mice","volume":"39","author":"Dietrich","year":"2014","journal-title":"J. Alzheimers Dis."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"e1586","DOI":"10.1038\/cddis.2014.553","article-title":"Tamoxifen reduces fat mass by boosting reactive oxygen species","volume":"6","author":"Liu","year":"2015","journal-title":"Cell Death Dis."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1038\/s41366-019-0330-3","article-title":"Even a low dose of tamoxifen profoundly induces adipose tissue browning in female mice","volume":"44","author":"Zhao","year":"2020","journal-title":"Int. J. Obes."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1016\/j.cmet.2011.08.009","article-title":"Distinct Hypothalamic Neurons Mediate Estrogenic Effects on Energy Homeostasis and Reproduction","volume":"14","author":"Xu","year":"2011","journal-title":"Cell Metab."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"117384","DOI":"10.1016\/j.lfs.2020.117384","article-title":"Beneficial effects of tamoxifen on leptin sensitivity in young mice fed a high fat diet: Role of estrogen receptor alpha and cytokines","volume":"246","author":"Farhadi","year":"2020","journal-title":"Life Sci."},{"key":"ref_51","unstructured":"Adam, M.P., Ardinger, H.H., Pagon, R.A., Wallace, S.E., Bean, L.J.H., Mirzaa, G., and Amemiya, A. (1993). Donnai-Barrow Syndrome. GeneReviews((R)), University of Washington."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"27459","DOI":"10.1038\/srep27459","article-title":"IVA cloning: A single-tube universal cloning system exploiting bacterial In Vivo Assembly","volume":"6","author":"Watson","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1093\/protein\/gzr058","article-title":"High-level secretion of recombinant monomeric murine and human single-chain Fv antibodies from Drosophila S2 cells","volume":"25","author":"Gilmartin","year":"2012","journal-title":"Protein Eng. Des. Sel."},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Backovic, M., and Krey, T. (2016). Stable Drosophila cell lines: An alternative approach to exogenous protein expression. Baculovirus and Insect Cell Expression Protocols, Springer.","DOI":"10.1007\/978-1-4939-3043-2_17"},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Harrison, L., Schriever, S.C., Feuchtinger, A., Kyriakou, E., Baumann, P., Pfuhlmann, K., Messias, A.C., Walch, A., Tschop, M.H., and Pfluger, P.T. (2018). Fluorescent blood-brain barrier tracing shows intact leptin transport in obese mice. Int. J. Obes.","DOI":"10.1038\/s41366-018-0221-z"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.brainres.2012.08.031","article-title":"An immortalised astrocyte cell line maintains the in vivo phenotype of a primary porcine in vitro blood-brain barrier model","volume":"1479","author":"Cantrill","year":"2012","journal-title":"Brain Res."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"101022","DOI":"10.1016\/j.molmet.2020.101022","article-title":"NF-kappaB signaling in tanycytes mediates inflammation-induced anorexia","volume":"39","author":"Sundaram","year":"2020","journal-title":"Mol. Metab."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"e2294","DOI":"10.21769\/BioProtoc.2294","article-title":"Isolation and Cultivation of Primary Brain Endothelial Cells from Adult Mice","volume":"7","author":"Assmann","year":"2017","journal-title":"Bio Protoc."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/9\/4998\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:58:15Z","timestamp":1760162295000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/9\/4998"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,5,8]]},"references-count":58,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2021,5]]}},"alternative-id":["ijms22094998"],"URL":"https:\/\/doi.org\/10.3390\/ijms22094998","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,5,8]]}}}