To access the full text documents, please follow this link:

Pro-inflammatory gene expression and neurotoxic effects of activated microglia are attenuated by absence of CCAAT/enhancer binding protein beta
Straccia, Marco; Gresa Arribas, Nuria; Dentesano, Guido; Ejarque Ortiz, Aroa; Tusell Puigbert, José Ma.; Serratosa i Serdà, Joan; Solà i Subirana, Carme; Saura Martí, Josep
Universitat de Barcelona
Background. Microglia and astrocytes respond to homeostatic disturbances with profound changes of gene expression. This response, known as glial activation or neuroinflammation, can be detrimental to the surrounding tissue. The transcription factor CCAAT/enhancer binding protein ß (C/EBPß) is an important regulator of gene expression in inflammation but little is known about its involvement in glial activation. To explore the functional role of C/EBPß in glial activation we have analyzed pro-inflammatory gene expression and neurotoxicity in murine wild type and C/EBPß-null glial cultures. Methods. Due to fertility and mortality problems associated with the C/EBPß-null genotype we developed a protocol to prepare mixed glial cultures from cerebral cortex of a single mouse embryo with high yield. Wild-type and C/EBPß-null glial cultures were compared in terms of total cell density by Hoechst-33258 staining; microglial content by CD11b immunocytochemistry; astroglial content by GFAP western blot; gene expression by quantitative real-time PCR, western blot, immunocytochemistry and Griess reaction; and microglial neurotoxicity by estimating MAP2 content in neuronal/microglial cocultures. C/EBPß DNA binding activity was evaluated by electrophoretic mobility shift assay and quantitative chromatin immunoprecipitation. Results. C/EBPß mRNA and protein levels, as well as DNA binding, were increased in glial cultures by treatment with lipopolysaccharide (LPS) or LPS + interferon ¿ (IFN¿). Quantitative chromatin immunoprecipitation showed binding of C/EBPß to pro-inflammatory gene promoters in glial activation in a stimulus- and gene-dependent manner. In agreement with these results, LPS and LPS+IFN¿ induced different transcriptional patterns between pro-inflammatory cytokines and NO synthase-2 genes. Furthermore, the expressions of IL-1ß and NO synthase-2, and consequent NO production, were reduced in the absence of C/EBPß. In addition, neurotoxicity elicited by LPS+IFN¿-treated microglia co-cultured with neurons was completely abolished by the absence of C/EBPß in microglia.
Expressió gènica
Gene expression
cc-by, (c) Straccia et al., 2011
BioMed Central

Show full item record

Related documents

Other documents of the same author

Dentesano, Guido; Straccia, Marco; Ejarque Ortiz, Aroa; Tusell Puigbert, José Ma; Serratosa i Serdà, Joan; Saura Martí, Josep; Solà i Subirana, Carme
Gresa Arribas, Núria; Viéitez, Cristina; Dentesano, Guido; Serratosa i Serdà, Joan; Saura Martí, Josep; Solà i Subirana, Carme
Pulido Salgado, Marta; Vidal Taboada, José Manuel; Garcia Diaz-Barriga, Gerardo; Serratosa i Serdà, Joan; Valente, Tony; Castillo, Paola; Matalonga, Jonathan; Straccia, Marco; Canals i Coll, Josep M.; Valledor Fernández, Annabel; Solà i Subirana, Carme; Saura Martí, Josep
Valente, Tony; Serratosa i Serdà, Joan; Perpiñá Martin, Unai; Saura Martí, Josep; Solà, Carme
Martínez Barriocanal, Águeda; Arcas García, Andrea; Magallon Lorenz, Miriam; Ejarque Ortiz, Aroa; Negro Demontel, M.Luciana; Comas Casellas, Emma; Schwartz Navarro, Simó; Malhotra, Sunny; Montalbán Gairín, Xavier; Peluffo, Hugo; Martín Andorrà, Margarita; Comabella López, Manuel; Sayós Ortega, Joan