Abstract
Existence of mitochondrial dysfunction is reported in the blood and brain of Parkinsonian patients. Cybrids are cytoplasmic hybrids created by the fusion of anucleate cells with rho0 (ρ0) cell line that has been deprived of its mitochondrial DNA. The present study describes production of ρ0 cells from SH-SY5Y cells and then uses these cells for the creation of normal and parkinsonian cybrids. Platelets from patients and age- and gender-matched controls were used to create cybrids by fusion of these platelets with the ρ0 cells. Further study involves their differentiation, characterization and comparison between control and Parkinson’s disease (PD) with specific consideration into the mitochondrial function or dysfunction in relation to the expression of certain nuclear and mitochondria encoded mitochondrial subunits. The expression in the proteins and their up- or down-regulation were extensively studied with the help of immunoblot and 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE). PD cybrids showed significant decline in the mitochondrial electron transport chain protein subunit expression (ND4, ND5 and ND6) with respect to the control cybrids. Twenty one mitochondrial proteins were differentially (up- or down- regulated) expressed as identified by 2-D PAGE followed by MALDI-TOF-TOF analysis. The platelets isolated from PD patient blood samples exhibited significantly lower complex I activities, as compared to control population. This, taken along with decreased mitochondrial complex-I subunit expression in the PD cybrids suggests the newly generated cell lines as a reliable cellular model of PD, and is an invaluable tool for mitochondrial research on this disease, and has been achieved for the first time in this part of the world. It is expected that the cybrids created in the laboratory would enable elucidation of mitochondrial machinery underlying PD pathology.
| Original language | American English |
|---|---|
| State | Published - Sep 2011 |