title: The Impact of CXCR4 Blockade on the Survival of Rat Brain Cortical Neurons.
abstract: BACKGROUND
  Chemokine receptor type 4 (CXCR4) plays a role in neuronal survival/cell repair and also contributes to the progression of cancer and neurodegenerative diseases. Chemokine ligand 12 (CXCL12) binds to CXCR4. In this study, we have investigated whether CXCR4 blockade by AMD3100 (a CXCR4 antagonist, member of bicyclam family) may affect neuronal survival in the absence of insult. Thus, we have measured the mitochondrial membrane potential (MMP), Bax and Bcl-2 protein translocation, and cytochrome c release in AMD3100-treated brain cortical neurons at 7 DIV (days in vitro).
  
  
  METHODS
  For this aim, AMD3100 (200 nM) was added to cortical neurons for 24 h, and several biomarkers like cell viability, reactive oxygen species (ROS) generation, lactate dehydrogenase (LDH) release, caspase-3/9 activity, proteins Bax and Bcl-2 translocation, and cytochromecrelease were analyzed by immunoblot.
  
  
  RESULTS
  CXCR4 blockade by AMD3100 (200 nM, 24 h) induces mitochondrial hyperpolarization and increases caspase-3/9 hyperpolarization without affecting LDH release as compared to untreated controls. AMD3100 also increases cytochrome c release and promotes Bax translocation to the mitochondria, whereas it raises cytosolic Bcl-2 levels in brain cortical neurons.
  
  
  CONCLUSIONS
  CXCR4 blockade induces cellular death via intrinsic apoptosis in rat brain cortical neurons in absence of insult.
authors.affiliation: Departamento de Bioquímica y Biología Molecular II, Facultad de Farmacia, Universidad Complutense de Madrid (U.C.M.), Ciudad Universitaria, 28040 Madrid, Spain. josem2005@yahoo.es.|Instituto de Investigación Neuroquímica (I.U.I.N.), Universidad Complutense de Madrid (U.C.M.), Ciudad Universitaria, 28040 Madrid, Spain. josem2005@yahoo.es.
authors.forename: José Joaquín
authors.initials: JJ
authors.lastname: Merino
authors.affiliation: Departamento de Farmacología, Facultad de Farmacia, Universidad Complutense de Madrid (U.C.M.), Ciudad Universitaria, 28040 Madrid, Spain. a.garcimartin@ucm.es.
authors.forename: Alba
authors.initials: A
authors.lastname: Garcimartín
authors.affiliation: Sección Departamental de Fisiología, Facultad de Farmacia, Universidad Complutense de Madrid (U.C.M.), Ciudad Universitaria, 28040 Madrid, Spain. elopez@ucm.es.
authors.forename: María Elvira
authors.initials: ME
authors.lastname: López-Oliva
authors.affiliation: Departamento de Farmacología, Facultad de Farmacia, Universidad Complutense de Madrid (U.C.M.), Ciudad Universitaria, 28040 Madrid, Spain. jbenedi@ucm.es.
authors.forename: Juana
authors.initials: J
authors.lastname: Benedí
authors.affiliation: Departamento de Bioquímica y Biología Molecular II, Facultad de Farmacia, Universidad Complutense de Madrid (U.C.M.), Ciudad Universitaria, 28040 Madrid, Spain. pilarg@ucm.es.
authors.forename: María Pilar
authors.initials: MP
authors.lastname: González
mesh_terms: D000818:Animals; D001596:Benzylamines; D015153:Blotting, Western; D001921:Brain / Q000166:cytology*; D016923:Cell Death / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D000080027:Cyclams; D006571:Heterocyclic Compounds / Q000494:pharmacology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D009474:Neurons / Q000166:cytology* / Q000187:drug effects*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D019718:Receptors, CXCR4 / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D001596:Benzylamines; C486264:Cxcr4 protein, rat; D000080027:Cyclams; D006571:Heterocyclic Compounds; D019253:Proto-Oncogene Proteins c-bcl-2; D017382:Reactive Oxygen Species; D019718:Receptors, CXCR4; D007770:L-Lactate Dehydrogenase; C088327:plerixafor
keywords: AMD3100; CXCR4 chemokine receptor; apoptosis; caspase-3/9; cortical neurons in vitro; cytochrome c/Bcl-2/Bax; cytosol; mitochondria; neuroprotection