Mitochondrial Overexpression
Mitochondrial Overexpression - Related Videos
15:43
Assessment of Mitochondrial Functions and Cell Viability in Renal Cells Overexpressing Protein Kinase C Isozymes
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Cited by 5 •
2013
The effects of activation of protein kinase C (PKC) isozymes on mitochondrial functions associated with respiration and oxidative phosphorylation and on cell viability are described. The approach adapts adenoviral technique to selectively overexpress PKC isozymes in primary cell culture and a variety of assays to determine mitochondrial functions and energy status of the cell.
02:59
Animal Mitochondrial Genetics
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2020
Among all the organelles in an animal cell, only mitochondria have their own independent genomes. Animal mitochondrial DNA is a double-stranded, closed-circular molecule with around 20,000 base pairs. Mitochondrial DNA is unique in that one of its two strands, the heavy, or H, -strand is guanine rich, whereas the complementary strand is cytosine rich and called the light, or L, -strand. Compared to nuclear DNA, mitochondrial DNA has a very low percentage of non-coding regions and is marked by...
08:40
An Optimized Protocol to Analyze Glycolysis and Mitochondrial Respiration in Lymphocytes
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Cited by 38 •
2016
The study of metabolism is becoming increasingly relevant to immunological research. Here, we present an optimized method for measuring glycolysis and mitochondrial respiration in mouse splenocytes, and T and B lymphocytes.
05:53
Mitochondrial Ca2+ Retention Capacity Assay and Ca2+-triggered Mitochondrial Swelling Assay
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Cited by 11 •
2018
This protocol aims to describe a method to examine the Ca2+ retention capacity and Ca2+- triggered mitochondrial swelling of isolated mitochondria of SH-SY5Y cells step-by-step.
05:16
Detecting Histone Modifications in Yeast Cells with Neurodegenerative Protein Overexpression
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2025
Source: Bennett, S. A., et al., Characterizing Histone Post-translational Modification Alterations in Yeast Neurodegenerative Proteinopathy Models. J. Vis. Exp. (2019).This video demonstrates the procedure of assessing histone modifications in yeast cells overexpressing neurodegenerative proteins. The steps include cell lysis, centrifugation, electrophoresis, membrane transfer, and antibody-based detection of modified histones.
FAQs
Mitochondrial-targeting sequences help direct newly produced proteins to the mitochondrion rather than leaving their distribution unspecified within the cell. This targeting is important when the research question concerns mitochondrial structure or function, because the intended protein must reach the organelle to influence processes such as electron transport, ATP production, or reactive oxygen species balance.
Increasing a mitochondrial factor can affect several connected outputs, including electron transport, ATP production, and the balance of reactive oxygen species. These changes may also influence cellular metabolism and stress responses. Measuring more than one outcome helps researchers determine whether the altered component has a focused effect or broader consequences for mitochondrial and cellular function.
Control cells provide the baseline needed to distinguish effects caused by increased mitochondrial expression from changes unrelated to the experimental manipulation. Comparing the two cell populations can reveal whether a factor alters mitochondrial structure, metabolism, stress responses, apoptosis, or disease-related pathways. Without this comparison, researchers cannot confidently attribute an observed outcome to the overexpressed component.
A typical strategy selects the mitochondrial protein or component of interest, introduces an engineered DNA construct, and uses a strong promoter to increase production. When organelle localization matters, the construct can also include a mitochondrial-targeting sequence. Researchers then compare the resulting cells with controls and examine relevant mitochondrial or cellular outcomes.
Useful outcomes include changes in mitochondrial structure and function, electron transport, ATP production, and reactive oxygen species balance. Researchers can also examine broader consequences for metabolism, cellular stress responses, apoptosis, and disease-related pathways. Considering these outcomes together connects a molecular change in a mitochondrial factor with its effects on cell biology.
This approach is useful when researchers need to test how a particular mitochondrial factor contributes to normal function or dysfunction. It supports investigations of metabolism, stress responses, apoptosis, and disease-related pathways, while also helping identify potential therapeutic targets. The comparison with control cells makes it possible to relate altered expression to specific biological consequences.
Raising the level of a selected mitochondrial component can expose links between that factor and disturbed electron transport, ATP production, reactive oxygen species balance, or cellular stress. Researchers can use these changes to investigate pathways associated with mitochondrial dysfunction and disease. Such experiments may also indicate whether the factor deserves consideration as a potential therapeutic target.