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Viability Assays for Cells in Culture
Published on: January 20, 2014
Different nitrosative-induced microtubular modifications and testosterone neuroprotective effects on
Sergio Gadau1, Gianluca Lepore, Marco Zedda
1Department of Animal Biology, University of Sassari, Italy. sgadau@uniss.it
Objectives:
Diabetic complications can often affect the central nervous system since the chronic exposure to hyperglycemia can result in the production of high concentration of reactive oxygen species with subsequent damage of several cell structures such as the cytoskeleton. In order to antagonize the oxidative status many substances have been tested as antioxidants. In the present work attention has been focused on the possible nitrosative effect of hyperglycemia on microtubular network of neuroblastoma and glioma mortalized cell lines, testing the possible neuroprotective effect of testosterone.
Methods:
Neuroblastoma (C1300) and glioma (C6) cell lines were cultured in the presence of 300 mM (C1300) or 310 mM (C6) D-glucose, with or without 50 nM testosterone. After 72 hrs, morphology, growth rate, cell viability and catalase activity were evaluated. In addition, with the aim to detect any changes in the amount of tubulin isoforms, Western blot analysis was performed.
Results:
In D-glucose-exposed cells, it was found a down-regulation of tubulin isoforms and an increase in 3-nitro-L-tyrosine and subsequent modifications in cell morphology, growth rate, viability and catalase activity. All these changes were more severe in neuroblastoma than in glioma cell line. When testosterone was added to the medium, all the parameters were very similar to controls. This neuroprotective action was well-detectable in C1300 cells, whereas testosterone was not able to recover significantly in C6 cells.
Conclusion:
Our results displayed: i) a selective action of high glucose on microtubules; ii) a different sensitivity to oxidative stress in neuronal and glial cells; iii) a different neuroprotective action of testosterone on neuronal and glial cells.
Insights
Testosterone protects neuroblastoma cells from high glucose-induced damage by preserving microtubule integrity. Glioma cells showed less response to testosterone, indicating differential neuroprotection in neuronal versus glial cells.
Area of Science:
- Neuroscience
- Cell Biology
- Endocrinology
Background:
- Diabetic complications can lead to central nervous system damage due to hyperglycemia-induced oxidative stress.
- Oxidative stress can damage cellular structures like the cytoskeleton, affecting neuronal function.
- Antioxidants are explored for their neuroprotective potential against diabetic complications.
Purpose of the Study:
- To investigate the nitrosative effects of hyperglycemia on the microtubular network in neuroblastoma and glioma cell lines.
- To evaluate the potential neuroprotective role of testosterone against hyperglycemia-induced damage.
Main Methods:
- Neuroblastoma (C1300) and glioma (C6) cells were exposed to high D-glucose concentrations with or without testosterone.
- Evaluated parameters included cell morphology, growth rate, viability, catalase activity, and tubulin isoform levels via Western blot.
Main Results:
- High glucose reduced tubulin levels and increased 3-nitro-L-tyrosine, altering cell morphology, growth, viability, and catalase activity.
- Neuroblastoma cells exhibited more severe damage than glioma cells.
- Testosterone treatment largely restored normal parameters in neuroblastoma cells but had limited effect on glioma cells.
Conclusions:
- High glucose selectively impacts microtubules, demonstrating differential sensitivity to oxidative stress between neuronal and glial cells.
- Testosterone exhibits a differential neuroprotective effect, more pronounced in neuronal (neuroblastoma) than glial (glioma) cells.
