Mitochondria Related Assay

Background of Mitochondria

The function of the nervous system heavily depends on mitochondria for energy supply. Mitochondria not only provide energy for cellular functions but are also involved in cellular signaling. When mitochondria are dysfunctional, the nervous system is often the first to be affected. Mitochondrial dysfunctions, such as abnormal mitochondria biogenesis and mitophagy as well as mitochondria damage, will lead to neurodegenerative diseases, including Parkinson's diseases, Alzheimer's, Huntington's disease, Friedreich's ataxia, and amyotrophic lateral sclerosis. Mitochondrial autophagy (i.e., mitophagy) is crucial to mitochondrial homeostasis, and mitophagy in the nervous system will decline with aging, which leads to the accumulation of dysfunctional mitochondria, oxidative stress-caused damage to mitochondria, and elevated cell apoptosis. Understanding the activation of mitochondria, mitochondrial membrane depolarization, mitochondrial stress, etc., is beneficial in developing new therapies to modulate mitochondrial biogenesis or activate mitophagy to remove impaired mitochondria, thereby minimizing the occurrence or severity of neurodegenerative diseases.

Fig.1 Mitochondrial dysfunction in neurodegenerative diseases. (Trombetta-Lima, et al., 2021)

Mitochondria Related Assay

So far, mitochondrial functions and activities can be tested and characterized by a series of assays, which include but are not limited to:

1. Neuronal mitochondrial activity assay can evaluate the effect of compounds on active mitochondria in soma versus neurites with imaging of MitoTracker Red labeled neurons.
2. Neuronal mitochondrial membrane depolarization assay can detect the capability of compounds to counteract mitochondrial membrane depolarization induced by selected lesion agents by comparing the fluorescence ratio of neuroblastoma cells to the control group.
3. Mitochondrial stress assay can be used to detect the compound effect on mitochondrial stress through the detection of oxygen consumption rates that are calculated from the changes in fluorescence signal using a fluorescence plate-reader over time.
4. Parkin mitochondrial recruitment assay is based on localization changes of two fluorescent proteins, which is used to analyze the effects of the compound on parkin recruitment and thus on mitochondrial integrity.
5. TOM20 represents a biomarker for mitochondrial abundance. TOM20 mitophagy assay combines data-rich high content analyses with high-throughput image acquisition to identify the effect of compounds on mitophagy with immunofluorescence assay.
6. Endoplasmic reticulum (ER)/Autophagic stress assay can be used to detect mitochondria stress.
7. Nuclear inclusion assay is a high-content assay that can measure the recruitment and accumulation of remodeled or dysfunctional mitochondria.

In vitro mitochondria-related assays offered by Creative Biolabs assists researchers in exploring the mechanisms of how mitochondria affect the nervous system. Researchers can thus develop novel therapies to target neurodegenerative diseases through our comprehensive platform. Please contact us if you want to learn more about in vitro mitochondria related assays. Our scientists are willing to answer any queries you may have regarding the relevant knowledge.

Reference

1. Trombetta-Lima, M.; et al. Mitochondrial dysfunction in neurodegenerative diseases: A focus on iPSC-derived neuronal models. Cell calcium. 2021, 94: 102362.