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Title: | Real-time PCR and immunocytochemical study of chondroitin sulfate proteoglycans after scratch wounding in cultured astrocytes | Authors: | Parabucki, Ana Santrač, Anja Savić, Danijela Dacić, Sanja Bjelobaba, Ivana Peković, Sanja Stojiljković, Mirjana |
Keywords: | chondroitin sulfate proteoglycans;gene expression;in vitro model;reactive astrocytes;scratch-wound assay | Issue Date: | 1-Oct-2013 | Journal: | Journal of Medical Biochemistry | Abstract: | Background: Various in vivo and in vitro models have been described in order to elucidate the pathobiology underlying the traumatic brain injury (TBI) and test potentially suitable treatments. Since TBI is a complex disease, models differ in regard to the aspect of TBI that is being investigated. One of the used in vitro models is the scratch wound assay, first established as a reproducible, low-cost assay for the analysis of cell migration in vitro. The aim of the present study was to further investigate the relevancy of this model as a counter- part of in vivo TBI models. Methods: We have examined the astrocytic response to a mechanical injury in terms of expression of chondroitin sul- fate proteoglycans (CSPGs) - phosphacan, neurocan and brevican, using real-time PCR and immunocytochemistry. Results: Our results indicate that in vitro scratch wounding alters the expression profile of examined CSPGs. Four hours after the scratch injury of the astrocytic monolayer, real-time PCR analysis revealed upregulation of mRNA levels for phos- phacan (3-fold) and neurocan (2-fold), whereas brevican mRNA was downregulated (2-fold). Immunofluorescent sig- nal for phosphacan and neurocan was more intense in astro- cytes close to the injury site, while brevican was scarcely present in cultured astrocytes. Conclusions: Obtained results indicate that CSPGs are differ- entially expressed by astrocytes after scratch wounding, demonstrating that the scratch wound model might be suit- able for investigation of astrocyte-derived response to injury. |
URI: | https://biore.bio.bg.ac.rs/handle/123456789/1583 | ISSN: | 1452-8258 | DOI: | 10.2478/jomb-2013-0036 |
Appears in Collections: | Journal Article |
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