title: Cultured cells of the blood-brain barrier from apolipoprotein B-100 transgenic mice: effects of oxidized low-density lipoprotein treatment.
abstract: BACKGROUND
  The apolipoprotein B-100 (ApoB-100) transgenic mouse line is a model of human atherosclerosis. Latest findings suggest the importance of ApoB-100 in the development of neurodegenerative diseases and microvascular/perivascular localization of ApoB-100 protein was demonstrated in the cerebral cortex of ApoB-100 transgenic mice. The aim of the study was to characterize cultured brain endothelial cells, pericytes and glial cells from wild-type and ApoB-100 transgenic mice and to study the effect of oxidized low-density lipoprotein (oxLDL) on these cells.
  
  
  METHODS
  Morphology of cells isolated from brains of wild type and ApoB-100 transgenic mice was characterized by immunohistochemistry and the intensity of immunolabeling was quantified by image analysis. Toxicity of oxLDL treatment was monitored by real-time impedance measurement and lactate dehydrogenase release. Reactive oxygen species and nitric oxide production, barrier permeability in triple co-culture blood-brain barrier model and membrane fluidity were also determined after low-density lipoprotein (LDL) or oxLDL treatment.
  
  
  RESULTS
  The presence of ApoB-100 was confirmed in brain endothelial cells, while no morphological change was observed between wild type and transgenic cells. Oxidized but not native LDL exerted dose-dependent toxicity in all three cell types, induced barrier dysfunction and increased reactive oxygen species (ROS) production in both genotypes. A partial protection from oxLDL toxicity was seen in brain endothelial and glial cells from ApoB-100 transgenic mice. Increased membrane rigidity was measured in brain endothelial cells from ApoB-100 transgenic mice and in LDL or oxLDL treated wild type cells.
  
  
  CONCLUSIONS
  The morphological and functional properties of cultured brain endothelial cells, pericytes and glial cells from ApoB-100 transgenic mice were characterized and compared to wild type cells for the first time. The membrane fluidity changes in ApoB-100 transgenic cells related to brain microvasculature indicate alterations in lipid composition which may be linked to the partial protection against oxLDL toxicity.
authors.affiliation: Laboratory of Animal Genetics and Molecular Neurobiology, Institute of Biochemistry, Biological Research Centre, Hungarian Academy of Sciences, Temesvári krt. 62, 6720, Szeged, Hungary. lenart.nikolett@koki.mta.hu.|Laboratory of Neuroimmunology, Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, Hungary. lenart.nikolett@koki.mta.hu.
authors.forename: Nikolett
authors.initials: N
authors.lastname: Lénárt
authors.affiliation: Biological Barriers Research Group, Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, Hungary. walter.fruzsina@brc.mta.hu.
authors.forename: Fruzsina R
authors.initials: FR
authors.lastname: Walter
authors.affiliation: Biological Barriers Research Group, Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, Hungary. bocsik.alexandra@brc.mta.hu.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Bocsik
authors.affiliation: Biological Barriers Research Group, Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, Hungary. santha.petra@brc.mta.hu.
authors.forename: Petra
authors.initials: P
authors.lastname: Sántha
authors.affiliation: Laboratory of Animal Genetics and Molecular Neurobiology, Institute of Biochemistry, Biological Research Centre, Hungarian Academy of Sciences, Temesvári krt. 62, 6720, Szeged, Hungary. toth.erzsebet.melinda@brc.mta.hu.
authors.forename: Melinda E
authors.initials: ME
authors.lastname: Tóth
authors.affiliation: Biological Barriers Research Group, Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, Hungary. harazin.andras@brc.mta.hu.
authors.forename: András
authors.initials: A
authors.lastname: Harazin
authors.affiliation: Biological Barriers Research Group, Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, Hungary. toth.andrea.e@gmail.com.
authors.forename: Andrea E
authors.initials: AE
authors.lastname: Tóth
authors.affiliation: Laboratory of Molecular Stress Biology, Institute of Biochemistry, Biological Research Centre, Hungarian Academy of Sciences, Szeged, Hungary. vizler.csaba@brc.mta.hu.
authors.forename: Csaba
authors.initials: C
authors.lastname: Vizler
authors.affiliation: Laboratory of Molecular Stress Biology, Institute of Biochemistry, Biological Research Centre, Hungarian Academy of Sciences, Szeged, Hungary. torok.zsolt@brc.mta.hu.
authors.forename: Zsolt
authors.initials: Z
authors.lastname: Török
authors.affiliation: Laboratory of Molecular Stress Biology, Institute of Biochemistry, Biological Research Centre, Hungarian Academy of Sciences, Szeged, Hungary. pilbat.ana_maria@brc.mta.hu.
authors.forename: Ana-Maria
authors.initials: AM
authors.lastname: Pilbat
authors.affiliation: Laboratory of Molecular Stress Biology, Institute of Biochemistry, Biological Research Centre, Hungarian Academy of Sciences, Szeged, Hungary. vigh.laszlo@brc.mta.hu.
authors.forename: László
authors.initials: L
authors.lastname: Vígh
authors.affiliation: Laboratory of Functional Genomics, Laboratories of Core Facilities, Biological Research Centre, Hungarian Academy of Sciences, Szeged, Hungary. laszlo@avidinbiotech.com.
authors.forename: László G
authors.initials: LG
authors.lastname: Puskás
authors.affiliation: Laboratory of Animal Genetics and Molecular Neurobiology, Institute of Biochemistry, Biological Research Centre, Hungarian Academy of Sciences, Temesvári krt. 62, 6720, Szeged, Hungary. santha.miklos@brc.mta.hu.
authors.forename: Miklós
authors.initials: M
authors.lastname: Sántha
authors.affiliation: Biological Barriers Research Group, Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, Hungary. deli.maria@brc.mta.hu.
authors.forename: Mária A
authors.initials: MA
authors.lastname: Deli
mesh_terms: D000818:Animals; D053299:Apolipoprotein B-100 / Q000235:genetics / Q000378:metabolism*; D050197:Atherosclerosis / Q000378:metabolism; D001812:Blood-Brain Barrier / Q000166:cytology* / Q000187:drug effects* / Q000502:physiology*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D008077:Lipoproteins, LDL / Q000633:toxicity*; D008560:Membrane Fluidity / Q000187:drug effects; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009457:Neuroglia / Q000187:drug effects / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D020286:Pericytes / Q000187:drug effects / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D053299:Apolipoprotein B-100; D008077:Lipoproteins, LDL; D017382:Reactive Oxygen Species; C099252:oxidized low density lipoprotein; D009569:Nitric Oxide