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Study of Protein Expression in Peri-Infarct Tissue after Cerebral Ischemia.

Brea D, Agulla J, Staes A, Gevaert K, Campos F, Sobrino T, Blanco M, Dávalos A, Castillo J, Ramos-Cabrer P - Sci Rep (2015)

Bottom Line: Apart from blood-related proteins (such as serum albumin), both techniques showed that the 70 kDa family of heat shock proteins were highly expressed in the peri-infarct tissue.Further studies by 1D and 2D western blotting and immunohistochemistry revealed that only one member of this family (the inducible form, HSP72 or HSP70i) is specifically expressed by the peri-infarct tissue, while the majority of this family (the constitutive form, HSC70 or HSP70c) is expressed in the whole brain.Our data support that HSP72 is a suitable biomarker of peri-infarct tissue in the ischemic brain.

View Article: PubMed Central - PubMed

Affiliation: 1] Neurology Department, Neurovascular Area, Clinical Neurosciences Research Laboratory, University Clinical Hospital, Health Research Institute of Santiago de Compostela (IDIS), University of Santiago de Compostela, Spain [2] Cellular and Molecular Neurobiology Research Group and Grup de Recerça en Neurociencies del IGTP, Department of Neurosciences, Fundació Institut d'Investigació en Ciències de la Salut Germans Trias I Pujol-Universitat Autónoma de Barcelona, Badalona, Spain.

ABSTRACT
In this work, we report our study of protein expression in rat peri-infarct tissue, 48 h after the induction of permanent focal cerebral ischemia. Two proteomic approaches, gel electrophoresis with mass spectrometry and combined fractional diagonal chromatography (COFRADIC), were performed using tissue samples from the periphery of the induced cerebral ischemic lesions, using tissue from the contra-lateral hemisphere as a control. Several protein spots (3408) were identified by gel electrophoresis, and 11 showed significant differences in expression between peri-infarct and contra-lateral tissues (at least 3-fold, p < 0.05). Using COFRADIC, 5412 proteins were identified, with 72 showing a difference in expression. Apart from blood-related proteins (such as serum albumin), both techniques showed that the 70 kDa family of heat shock proteins were highly expressed in the peri-infarct tissue. Further studies by 1D and 2D western blotting and immunohistochemistry revealed that only one member of this family (the inducible form, HSP72 or HSP70i) is specifically expressed by the peri-infarct tissue, while the majority of this family (the constitutive form, HSC70 or HSP70c) is expressed in the whole brain. Our data support that HSP72 is a suitable biomarker of peri-infarct tissue in the ischemic brain.

No MeSH data available.


Related in: MedlinePlus

Schematic representation of the experimental procedures performed.Three tissue portions were isolated from rat brains at 48 h post-ischemia: infarcted tissue (lesion), as delimited by TTC staining; a 2-mm strip around the infarct region (peri-infarct); and control tissue from the contra-lateral hemisphere. The protein content from each portion was extracted in three different fractions (membrane, soluble and insoluble). The samples were analysed 2D-PAGE and COFRADIC proteomics. Proteins with significant differences of expression between the tissue portions (>3-fold, p < 0.05) were further studied by western blotting (WB) and immunohistochemistry (IHC). Data were combined to define suitable molecular targets of the peri-infarct tissue.
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f1: Schematic representation of the experimental procedures performed.Three tissue portions were isolated from rat brains at 48 h post-ischemia: infarcted tissue (lesion), as delimited by TTC staining; a 2-mm strip around the infarct region (peri-infarct); and control tissue from the contra-lateral hemisphere. The protein content from each portion was extracted in three different fractions (membrane, soluble and insoluble). The samples were analysed 2D-PAGE and COFRADIC proteomics. Proteins with significant differences of expression between the tissue portions (>3-fold, p < 0.05) were further studied by western blotting (WB) and immunohistochemistry (IHC). Data were combined to define suitable molecular targets of the peri-infarct tissue.

Mentions: The experimental procedure used in this study is schematically represented in Fig. 1. Brain sections from animals subjected to a focal permanent cerebral ischemia were subsequently studied by proteomics, blotting and immunohistochemistry techniques, as described in the methods section.


Study of Protein Expression in Peri-Infarct Tissue after Cerebral Ischemia.

Brea D, Agulla J, Staes A, Gevaert K, Campos F, Sobrino T, Blanco M, Dávalos A, Castillo J, Ramos-Cabrer P - Sci Rep (2015)

Schematic representation of the experimental procedures performed.Three tissue portions were isolated from rat brains at 48 h post-ischemia: infarcted tissue (lesion), as delimited by TTC staining; a 2-mm strip around the infarct region (peri-infarct); and control tissue from the contra-lateral hemisphere. The protein content from each portion was extracted in three different fractions (membrane, soluble and insoluble). The samples were analysed 2D-PAGE and COFRADIC proteomics. Proteins with significant differences of expression between the tissue portions (>3-fold, p < 0.05) were further studied by western blotting (WB) and immunohistochemistry (IHC). Data were combined to define suitable molecular targets of the peri-infarct tissue.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4495553&req=5

f1: Schematic representation of the experimental procedures performed.Three tissue portions were isolated from rat brains at 48 h post-ischemia: infarcted tissue (lesion), as delimited by TTC staining; a 2-mm strip around the infarct region (peri-infarct); and control tissue from the contra-lateral hemisphere. The protein content from each portion was extracted in three different fractions (membrane, soluble and insoluble). The samples were analysed 2D-PAGE and COFRADIC proteomics. Proteins with significant differences of expression between the tissue portions (>3-fold, p < 0.05) were further studied by western blotting (WB) and immunohistochemistry (IHC). Data were combined to define suitable molecular targets of the peri-infarct tissue.
Mentions: The experimental procedure used in this study is schematically represented in Fig. 1. Brain sections from animals subjected to a focal permanent cerebral ischemia were subsequently studied by proteomics, blotting and immunohistochemistry techniques, as described in the methods section.

Bottom Line: Apart from blood-related proteins (such as serum albumin), both techniques showed that the 70 kDa family of heat shock proteins were highly expressed in the peri-infarct tissue.Further studies by 1D and 2D western blotting and immunohistochemistry revealed that only one member of this family (the inducible form, HSP72 or HSP70i) is specifically expressed by the peri-infarct tissue, while the majority of this family (the constitutive form, HSC70 or HSP70c) is expressed in the whole brain.Our data support that HSP72 is a suitable biomarker of peri-infarct tissue in the ischemic brain.

View Article: PubMed Central - PubMed

Affiliation: 1] Neurology Department, Neurovascular Area, Clinical Neurosciences Research Laboratory, University Clinical Hospital, Health Research Institute of Santiago de Compostela (IDIS), University of Santiago de Compostela, Spain [2] Cellular and Molecular Neurobiology Research Group and Grup de Recerça en Neurociencies del IGTP, Department of Neurosciences, Fundació Institut d'Investigació en Ciències de la Salut Germans Trias I Pujol-Universitat Autónoma de Barcelona, Badalona, Spain.

ABSTRACT
In this work, we report our study of protein expression in rat peri-infarct tissue, 48 h after the induction of permanent focal cerebral ischemia. Two proteomic approaches, gel electrophoresis with mass spectrometry and combined fractional diagonal chromatography (COFRADIC), were performed using tissue samples from the periphery of the induced cerebral ischemic lesions, using tissue from the contra-lateral hemisphere as a control. Several protein spots (3408) were identified by gel electrophoresis, and 11 showed significant differences in expression between peri-infarct and contra-lateral tissues (at least 3-fold, p < 0.05). Using COFRADIC, 5412 proteins were identified, with 72 showing a difference in expression. Apart from blood-related proteins (such as serum albumin), both techniques showed that the 70 kDa family of heat shock proteins were highly expressed in the peri-infarct tissue. Further studies by 1D and 2D western blotting and immunohistochemistry revealed that only one member of this family (the inducible form, HSP72 or HSP70i) is specifically expressed by the peri-infarct tissue, while the majority of this family (the constitutive form, HSC70 or HSP70c) is expressed in the whole brain. Our data support that HSP72 is a suitable biomarker of peri-infarct tissue in the ischemic brain.

No MeSH data available.


Related in: MedlinePlus