The Association of VDAC with Cell Viability of PC12 Model of Huntington's Disease
Andonis Karachitos1, Daria Grobys1, Klaudia Kulczyńska1
1Laboratory of Bioenergetics, Faculty of Biology, Institute of Molecular Biology and Biotechnology, Adam Mickiewicz University in Poznań , Poznań , Poland.
Insights
Mutant huntingtin (mHtt) expression alters voltage-dependent anion-selective channel (VDAC) function, impacting mitochondria and cell survival in Huntington
Area of Science:
- Cell Biology
- Neuroscience
- Mitochondrial Biology
Background:
- Mitochondrial dysfunction is implicated in Huntington's disease (HD) pathogenesis.
- The precise mechanisms linking mitochondria dysfunction to cell death in HD remain unclear.
- Voltage-dependent anion-selective channel (VDAC) plays a critical role in mitochondrial function.
Purpose of the Study:
- To investigate the impact of wild-type huntingtin (Htt) and mutant huntingtin (mHtt) expression on cell survival and mitochondrial function.
- To determine the effect of Htt and mHtt expression on VDAC function in an inducible PC12 cell model of HD.
Main Methods:
- Utilized an inducible PC12 cell model expressing Htt and mHtt.
- Assessed cell viability and mitochondrial function after 48 hours of protein expression.
- Isolated VDAC and analyzed its function using a black lipid membrane system.
Main Results:
- Expression of mHtt, but not Htt, altered VDAC's open state conductance and voltage-dependence.
- VDAC1 was the dominant VDAC isoform, and its relative abundance was unchanged by Htt/mHtt expression.
- Functional VDAC changes occurred shortly after Htt and mHtt expression.
Conclusions:
- Htt and mHtt expression-induced functional alterations in VDAC, primarily VDAC1, contribute to mitochondrial dysfunction and reduced cell viability in HD models.
- These findings offer insights into HD cytotoxicity and potential cytoprotective strategies.
Abstract:
It is becoming increasingly apparent that mitochondria dysfunction plays an important role in the pathogenesis of Huntington's disease (HD), but the underlying mechanism is still elusive. Thus, there is a still need for further studies concerning the upstream events in the mitochondria dysfunction that could contribute to cell death observed in HD. Taking into account the fundamental role of the voltage-dependent anion-selective channel (VDAC) in mitochondria functioning, it is reasonable to consider the channel as a crucial element in HD etiology. Therefore, we applied inducible PC12 cell model of HD to determine the relationship between the effect of expression of wild type and mutant huntingtin (Htt and mHtt, respectively) on cell survival and mitochondria functioning in intact cells under conditions of undergoing cell divisions. Because after 48 h of Htt and mHtt expression differences in mitochondria functioning co-occurred with differences in the cell viability, we decided to estimate the effect of Htt and mHtt expression lasted for 48 h on VDAC functioning. Therefore, we isolated VDAC from the cells and tested the preparations by black lipid membrane system. We observed that the expression of mHtt, but not Htt, resulted in changes of the open state conductance and voltage-dependence when compared to control cells cultured in the absence of the expression. Importantly, for all the VDAC preparations, we observed a dominant quantitative content of VDAC1, and the quantitative relationships between VDAC isoforms were not changed by Htt and mHtt expression. Thus, Htt and mHtt-mediated functional changes of VDAC, being predominantly VDAC1, which occur shortly after these protein appearances in cells, may result in differences concerning mitochondria functioning and viability of cells expressing Htt and mHtt. The assumption is important for better understanding of cytotoxicity as well as cytoprotection mechanisms of potential clinical application.
