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Title

Generation of LyzM-Cre x C/EBPβfl/fl mice to obtain selective depletion of the transcription factor C/EBPβ in microglia

AuthorsPulido-Salgado, Marta ; Straccia, Marco ; Sterneck, E.; Solà, Carme ; Saura, Josep
Issue DateJul-2012
CitationI CIBICAT (2012)
AbstractComplete deficiency of the transcription factor CCAAT/enhancer binding protein β (C/EBPβ) protects from excitotoxic and ischemic insults suggesting the involvement of C/EBPβ in neurotoxicity in these models. C/EBPβ can be expressed by numerous cell types and the cell types involved in the potential detrimental effects of C/EBPβ have not been determined. We have recently shown that C/EBPβ absence results in the attenuated expression of pro-inflammatory genes in primary mixed glial cultures and in the abrogation of the neurotoxicity elicited by activated microglia in neuron-microglia co-cultures. In order to determine the neuroprotective potential of C/EBPβ inhibition in microglia we have generated a mouse transgenic line in which C/EBPβ gene is flanked by LoxP sequences (C/EBPβfl/fl) and Cre-recombinase is expressed under the control of the microglia/macrophage promoter Lysozyme M. In primary mixed glial cultures from wild-type or from the two parental lines, C/EBPβ was expressed both in astrocytes and microglia. Primary mixed glial cultures prepared from the cerebral cortex of neonatal LyzM-Cre x C/EBPβfl/fl mice revealed that virtually all microglial cells, identified by CD11b or Iba1 immunostaining, lacked C/EBPβ expression whereas C/EBPβ expression in astrocytes, identified by GFAP immunostaining, was similar to control lines. Interestingly, LPS or LPS+IFNγ-induced NO production in mixed glial cultures, which is of microglial origin, was markedly attenuated in LyzM-Cre x C/EBPβfl/fl cultures when compared to control lines and the same effect was observed in microglial-enriched cultures. We are in the process of obtaining samples to analyze gene expression profile of control and activated microglial cultures from LyzM-Cre x C/EBPβfl/fl and wild-type mice. In parallel, we are interested to determine whether the cell specific absence of C/EBPβ in microglia in LyzM-Cre x C/EBPβfl/fl mice is also observed in vivo. In this case, LyzM-Cre x C/EBPβfl/fl mice would be a useful tool to analyze the involvement of microglial C/EBPβ in neurodegeneration in vivo and to test the hypothesis that inhibition of microglial C/EBPβ could have potential for the treatment of neurological disorders in which neuroinflammation plays a pathogenic role.
DescriptionTrabajo presentado al I Congrés Internacional de Biologia de Catalunya: Global questions on advanced biology, celebrado en Barcelona del 9 al 12 de julio de 2012.
URIhttp://hdl.handle.net/10261/92435
Appears in Collections:(IIBB) Comunicaciones congresos
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