Técnicas Avançadas em Biologia e Medicina
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Isolation of Mitochondrial RNA with 6-OHDA Treatment to Detect Cell Viability of HtrA2 KO/CHOP Genotype

Al-Baraa Akram El-Sayed*

Mitochondrial RNAs are a class of small RNAs which can regulate gene expression through interference of translation process. A particular class of these miRNAs was found to be enriched in proximity of mitochondria and inside these organelles controlling the behavior of the mitochondria. The RNA in mitochondria is organized by special and membrane less compartments of RNA-protein complexes, called the RNA mitochondrial granules, which are abbreviated into MRGs.

Most methods used to isolate mitochondria depends on differential centrifugation, a two-step centrifugation carried out at low speed to remove intact cells, cell and tissue debris as well as nuclei from whole cell extracts followed by high speed centrifugation to concentrate mitochondria and separate mitochondria from other cellular organelles. Molecular quality control marker Hsp60 and organ-cellular quality control marker Atg5. Drug treatment with 6-OHDA induced decreased cell viability and ATP production of HtrA2 KO/CHOP genotype. Displaying a dmg concentration-dependent effect which reflects both the cell viability and the metabolic status of the cells. Drug treatment with 6-OHDA in conjunction with ADEP4 or ACP5 significantly improved cell survival, ATP production. Indicating that these drugs might have a protective effect while reoccurring the detrimental effects or 6-OHDA. The findings provide new evidence that depiction of HtrA2 might contribute to an increase in mitochondrial stress and transcriptional up regulation of the nuclear stress related to CHOP gene. Both are involved in Parkinson's disease.