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Assessment of Protein Expression Levels After Transient Global Cerebral Ischemia Using an Antibody Microarray Analysis

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Abstract

An antibody microarray was used to analyze potential modifications in brain protein levels induced by ischemic reperfusion. Total brain extracts from rats subjected to 15 min of transient global ischemia followed by 3 days of reperfusion and sham control animals were compared within the same array. Separate arrays were run to analyze resistant (cortex) and vulnerable (CA1) regions to ischemia. Candidate components distinguishing the two cellular populations were selected under stringent criteria. IR significantly decreased the expression of Bcl-x, caspase 11, GADD153, Cdk4, E2F1, Retinoblastoma-P, SMAD4, AP-1/c-jun, ATF2, PCAF, MAP1b and cofilin within both regions. NGF and NMDA 2A receptors and IκB were specifically down-regulated in CA1, while Pyk2-P, b-NOS, and tyrosine hydroxylase were slightly up-regulated in the same region. Some of the array results were validated by western blot. Both the array and western blot results suggested a relevant IR induced activation of calpain specifically at CA1.

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Acknowledgments

This work was supported by grants from the Instituto de Salud Carlos III, Spain: 08/0761; 06/0289 and RETICS-RD06/0026/008, including a predoctoral position for MIA. We thank Mercedes Gomez-Calcerrada for her technical assistance. MEM is a researcher from FIBio-HRC supported by Consejeria de Sanidad (Comunidad de Madrid), Spain.

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Authors and Affiliations

  1. Servicio de Bioquímica-Investigación, Hospital Ramón y Cajal, Irycis, Ctra. Colmenar Km.9, 28034, Madrid, Spain

    Maria Irene Ayuso, Maria Elena Martín & Matilde Salinas

  2. Neurovascular Research Laboratory, Neurovascular Unit, Institut de Recerca, Hospital Vall d’Hebron, Barcelona, Spain

    Lidia García-Bonilla

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  1. Maria Irene Ayuso
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  2. Lidia García-Bonilla
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  3. Maria Elena Martín
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  4. Matilde Salinas
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Correspondence to Matilde Salinas.

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Ayuso, M.I., García-Bonilla, L., Martín, M.E. et al. Assessment of Protein Expression Levels After Transient Global Cerebral Ischemia Using an Antibody Microarray Analysis. Neurochem Res 35, 1239–1247 (2010). https://doi.org/10.1007/s11064-010-0180-9

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  • DOI: https://doi.org/10.1007/s11064-010-0180-9

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