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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

Immunohistochemistry of two consecutive slices of brain tissue showing the expression of HSP70 proteins.Images (4x and 10x) showing the expression of the constitutive form of HSP70 (HSP70 or HSP70c) in cells of the infarct, peri-infarct, and contralateral areas of an ischemic brain (top row), while the inducible form of this protein (HSP72 or HSP70i) is only expressed in cells of the peri-infarct region, and not in the other two regions (lower row).
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f5: Immunohistochemistry of two consecutive slices of brain tissue showing the expression of HSP70 proteins.Images (4x and 10x) showing the expression of the constitutive form of HSP70 (HSP70 or HSP70c) in cells of the infarct, peri-infarct, and contralateral areas of an ischemic brain (top row), while the inducible form of this protein (HSP72 or HSP70i) is only expressed in cells of the peri-infarct region, and not in the other two regions (lower row).

Mentions: Immunolabelling of brain sections was performed with an antibody that specifically labelled HSP72 and with another antibody that labelled both the inducible and constitutive forms of HSP70 (Fig. 5). While HSP72 expression was not observed in the contra-lateral tissue and was nearly absent in the lesion site, the expression of this protein is clearly visible in the peri-infarct region. On the contrary, strong staining for the antibody against the constitutive form of HSP70 was observed in both the peri-infarct and the contra-lateral (control) tissue.


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)

Immunohistochemistry of two consecutive slices of brain tissue showing the expression of HSP70 proteins.Images (4x and 10x) showing the expression of the constitutive form of HSP70 (HSP70 or HSP70c) in cells of the infarct, peri-infarct, and contralateral areas of an ischemic brain (top row), while the inducible form of this protein (HSP72 or HSP70i) is only expressed in cells of the peri-infarct region, and not in the other two regions (lower row).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f5: Immunohistochemistry of two consecutive slices of brain tissue showing the expression of HSP70 proteins.Images (4x and 10x) showing the expression of the constitutive form of HSP70 (HSP70 or HSP70c) in cells of the infarct, peri-infarct, and contralateral areas of an ischemic brain (top row), while the inducible form of this protein (HSP72 or HSP70i) is only expressed in cells of the peri-infarct region, and not in the other two regions (lower row).
Mentions: Immunolabelling of brain sections was performed with an antibody that specifically labelled HSP72 and with another antibody that labelled both the inducible and constitutive forms of HSP70 (Fig. 5). While HSP72 expression was not observed in the contra-lateral tissue and was nearly absent in the lesion site, the expression of this protein is clearly visible in the peri-infarct region. On the contrary, strong staining for the antibody against the constitutive form of HSP70 was observed in both the peri-infarct and the contra-lateral (control) tissue.

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