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Changes in HSP gene and protein expression in natural scrapie with brain damage.

Serrano C, Bolea R, Lyahyai J, Filali H, Varona L, Marcos-Carcavilla A, Acín C, Calvo JH, Serrano M, Badiola JJ, Zaragoza P, Martín-Burriel I - Vet. Res. (2011)

Bottom Line: Whereas changes in transcript levels were not observed in the cerebellum or medulla oblongata, a significant decrease in HSP27 and HSP90 was detected in the prefrontal cortex.Changes in Hsp gene and protein expression were associated with prion protein deposition, gliosis and spongiosis rather than with apoptosis.Finally, immunohistochemistry revealed intense Hsp70 and Hsp90 immunolabelling in Purkinje cells of scrapie sheep.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratorio de Genética Bioquímica (LAGENBIO), Facultad de Veterinaria, Universidad de Zaragoza, Miguel Servet 177, 50013 Zaragoza, Spain. minma@unizar.es.

ABSTRACT
Heat shock proteins (Hsp) perform cytoprotective functions such as apoptosis regulation and inflammatory response control. These proteins can also be secreted to the extracellular medium, acting as inflammatory mediators, and their chaperone activity permits correct folding of proteins and avoids the aggregation of anomalous isoforms. Several studies have proposed the implication of Hsp in prion diseases. We analysed the gene expression and protein distribution of different members of the Hsp27, Hsp70, and Hsp90 families in the central nervous system of sheep naturally infected with scrapie. Different expression profiles were observed in the areas analysed. Whereas changes in transcript levels were not observed in the cerebellum or medulla oblongata, a significant decrease in HSP27 and HSP90 was detected in the prefrontal cortex. In contrast, HSP73 was over-expressed in diencephalons of scrapie animals. Western blotting did not reveal significant differences in Hsp90 and Hsp70 protein expression between scrapie and control animals. Expression rates identified by real-time RT-PCR and western blotting were compared with the extent of classical scrapie lesions using stepwise regression. Changes in Hsp gene and protein expression were associated with prion protein deposition, gliosis and spongiosis rather than with apoptosis. Finally, immunohistochemistry revealed intense Hsp70 and Hsp90 immunolabelling in Purkinje cells of scrapie sheep. In contrast, controls displayed little or no staining in these cells. The observed differences in gene expression and protein distribution suggest that the heat shock proteins analysed play a role in the natural form of the disease.

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Hsp protein expression and distribution in scrapie tissues. Specificity of anti-Hsp90 and anti-Hsp70 antibodies in the ovine medulla oblongata, as detected by western blotting (a). Arrowheads indicate intense Hsp70 immunostaining of spheroids observed in medulla oblongata (b). Immunohistochemical determination of Hsp70 in scrapie (c) and control (d) animals; Hsp70-positive and -negative Purkinje cells are indicated with arrows and arrowheads, respectively. Immunohistochemical determination of Hsp90 in scrapie (e) and control (f) animals; Hsp90-negative Purkinje cells are indicated by arrowheads, and Hsp90-positive cells are indicated by arrows. Significant increase in the percentage of Purkinje cells staining positively for Hsp70 (g) and Hsp90 (h) in scrapie cerebella. ** p < 0.01 (Student's t-test).
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Figure 2: Hsp protein expression and distribution in scrapie tissues. Specificity of anti-Hsp90 and anti-Hsp70 antibodies in the ovine medulla oblongata, as detected by western blotting (a). Arrowheads indicate intense Hsp70 immunostaining of spheroids observed in medulla oblongata (b). Immunohistochemical determination of Hsp70 in scrapie (c) and control (d) animals; Hsp70-positive and -negative Purkinje cells are indicated with arrows and arrowheads, respectively. Immunohistochemical determination of Hsp90 in scrapie (e) and control (f) animals; Hsp90-negative Purkinje cells are indicated by arrowheads, and Hsp90-positive cells are indicated by arrows. Significant increase in the percentage of Purkinje cells staining positively for Hsp70 (g) and Hsp90 (h) in scrapie cerebella. ** p < 0.01 (Student's t-test).

Mentions: A single band of ~70 kDa, approximately, was obtained by Western blotting, demonstrating the specificity of the anti-Hsp70 antibody (Figure 2). No statistically significant changes were observed between controls and scrapie-affected individuals with respect to global Hsp70 expression in the four CNS zones. Using the anti-Hsp90 antibody, Western blot also revealed a single band that corresponded to the expected value (~90 kDa) (Figure 2). As for Hsp70, no significant differences in Hsp90 expression were detected between controls and scrapie-affected animals in any area of the CNS.


Changes in HSP gene and protein expression in natural scrapie with brain damage.

Serrano C, Bolea R, Lyahyai J, Filali H, Varona L, Marcos-Carcavilla A, Acín C, Calvo JH, Serrano M, Badiola JJ, Zaragoza P, Martín-Burriel I - Vet. Res. (2011)

Hsp protein expression and distribution in scrapie tissues. Specificity of anti-Hsp90 and anti-Hsp70 antibodies in the ovine medulla oblongata, as detected by western blotting (a). Arrowheads indicate intense Hsp70 immunostaining of spheroids observed in medulla oblongata (b). Immunohistochemical determination of Hsp70 in scrapie (c) and control (d) animals; Hsp70-positive and -negative Purkinje cells are indicated with arrows and arrowheads, respectively. Immunohistochemical determination of Hsp90 in scrapie (e) and control (f) animals; Hsp90-negative Purkinje cells are indicated by arrowheads, and Hsp90-positive cells are indicated by arrows. Significant increase in the percentage of Purkinje cells staining positively for Hsp70 (g) and Hsp90 (h) in scrapie cerebella. ** p < 0.01 (Student's t-test).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3037893&req=5

Figure 2: Hsp protein expression and distribution in scrapie tissues. Specificity of anti-Hsp90 and anti-Hsp70 antibodies in the ovine medulla oblongata, as detected by western blotting (a). Arrowheads indicate intense Hsp70 immunostaining of spheroids observed in medulla oblongata (b). Immunohistochemical determination of Hsp70 in scrapie (c) and control (d) animals; Hsp70-positive and -negative Purkinje cells are indicated with arrows and arrowheads, respectively. Immunohistochemical determination of Hsp90 in scrapie (e) and control (f) animals; Hsp90-negative Purkinje cells are indicated by arrowheads, and Hsp90-positive cells are indicated by arrows. Significant increase in the percentage of Purkinje cells staining positively for Hsp70 (g) and Hsp90 (h) in scrapie cerebella. ** p < 0.01 (Student's t-test).
Mentions: A single band of ~70 kDa, approximately, was obtained by Western blotting, demonstrating the specificity of the anti-Hsp70 antibody (Figure 2). No statistically significant changes were observed between controls and scrapie-affected individuals with respect to global Hsp70 expression in the four CNS zones. Using the anti-Hsp90 antibody, Western blot also revealed a single band that corresponded to the expected value (~90 kDa) (Figure 2). As for Hsp70, no significant differences in Hsp90 expression were detected between controls and scrapie-affected animals in any area of the CNS.

Bottom Line: Whereas changes in transcript levels were not observed in the cerebellum or medulla oblongata, a significant decrease in HSP27 and HSP90 was detected in the prefrontal cortex.Changes in Hsp gene and protein expression were associated with prion protein deposition, gliosis and spongiosis rather than with apoptosis.Finally, immunohistochemistry revealed intense Hsp70 and Hsp90 immunolabelling in Purkinje cells of scrapie sheep.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratorio de Genética Bioquímica (LAGENBIO), Facultad de Veterinaria, Universidad de Zaragoza, Miguel Servet 177, 50013 Zaragoza, Spain. minma@unizar.es.

ABSTRACT
Heat shock proteins (Hsp) perform cytoprotective functions such as apoptosis regulation and inflammatory response control. These proteins can also be secreted to the extracellular medium, acting as inflammatory mediators, and their chaperone activity permits correct folding of proteins and avoids the aggregation of anomalous isoforms. Several studies have proposed the implication of Hsp in prion diseases. We analysed the gene expression and protein distribution of different members of the Hsp27, Hsp70, and Hsp90 families in the central nervous system of sheep naturally infected with scrapie. Different expression profiles were observed in the areas analysed. Whereas changes in transcript levels were not observed in the cerebellum or medulla oblongata, a significant decrease in HSP27 and HSP90 was detected in the prefrontal cortex. In contrast, HSP73 was over-expressed in diencephalons of scrapie animals. Western blotting did not reveal significant differences in Hsp90 and Hsp70 protein expression between scrapie and control animals. Expression rates identified by real-time RT-PCR and western blotting were compared with the extent of classical scrapie lesions using stepwise regression. Changes in Hsp gene and protein expression were associated with prion protein deposition, gliosis and spongiosis rather than with apoptosis. Finally, immunohistochemistry revealed intense Hsp70 and Hsp90 immunolabelling in Purkinje cells of scrapie sheep. In contrast, controls displayed little or no staining in these cells. The observed differences in gene expression and protein distribution suggest that the heat shock proteins analysed play a role in the natural form of the disease.

Show MeSH
Related in: MedlinePlus