C. elegans Oxygen Consumption Rate Measurement Service
Mitochondrial function is critical for organismal health and refers to various physiological activities, including energy production, reactive oxygen species (ROS) scavenging, calcium homeostasis, and apoptosis. In particular, for cells that have high energy demands, like neurocytes and muscle cells, having an optimal mitochondrial function is extremely important. Consistent with this, mitochondrial dysfunction has been connected with plenty of aging and neurodegenerative diseases. Mitochondrial function can be determined using multiple assays, involving in measuring ATP and ROS levels, mitochondrial membrane potential and mitochondrial calcium concentrations, while these assays only elucidate mitochondrial function from a single snapshot. It is reported that mitochondrial dysfunction can induce variations of oxygen consumption, which depend on a broad range of sequential reactions during ATP generation. Therefore, mitochondrial respiration is a superior indicator of mitochondrial function that may provide a comprehensive view of mitochondrial health. And the measurement of oxygen consumption rate (OCR) is highly valuable in a biological system and can be conducted both in in vitro and in vivo assays.
Respiration of C. elegans
As an intact animal with functional systems, the nematode has many advantages, such as small in size, easy to maintain, and especially maintained axenically. Besides, studies have performed prediction of mammalian development and the cost is a fraction of the traditional animal testing. Therefore, C. elegans has been extensively exploited as a powerful model system for high-throughput assessment protocols, referring to drugs, pollutants, and other environmental samples. For C. elegans, the commonly applied toxicity endpoints involve survival, reproduction and behaviors. Among the indicators, respiration of C. elegans is not frequently used to study the effect of toxicant exposure. However, the endpoint has been indicated to serve as an effective toxic measurement, especially in the aspect of metal pollutions.
Service offerings
OCR is a measurement of respiration, and the relevance and applicability of which have been greatly improved with the state-of-the-art development. Here we offer Seahorse XFe96 Extracellular Flux Analyzer, a high-throughput respirometer, to measure the oxygen consumption rate in C. elegans. Despite the Seahorse respirometer setup being originally designed to deal with ex vivo cells, it can also be utilized in vivo with small living beings like C. elegans. Furthermore, it is notable that the setup has facilitated the development of acute response protocols that measure C. elegans OCR.
The service described in the following sections
Schematic overview of C. elegans OCR measurement using
the Seahorse XFe96 Extracellular Flux Analyzer.( Preez GD, et al, 2020)
- Stock preparation of the food source (Escherichia coli)
Food availability has a substantial impact on C. elegans development. Apart from the important step to prepare and store the food stock, a food density bioassay is conducted.
- Growth and synchronization of C. elegans
- Preparation of toxicant solution and hydration of probes
- Preparation of nematode plate and run assays.
The bioassay is performed on 96-well culture plates, or bioassay plates. The purpose of bioassay plate layout is to allow the access of the maximum number of exposure concentrations with sufficient replication.
- Post-experiment data analysis
- Food density bioassay
- Toxicant concentration-response bioassays
- Oxygen consumption rate response to temperature fluctuations
CD BioSciences is a professional C. elegans model service provider. Caenorhabditis elegans is a simple but powerful model organism, which can be used to elucidate the many intricacies of mitochondrial function. Mitochondrial respiration is an insightful indicator of mitochondrial function and here we provide the state-of-the-art platform to measure respiration in C. elegans using developed respirometers Seahorse XFe96 Extracellular Flux Analyzer. If you are interested in our services, please don't hesitate to contact us.
Reference
- Preez GD, et al. (2020) "Oxygen consumption rate of Caenorhabditis elegans as a high-throughput endpoint of toxicity testing using the Seahorse XFe96 Extracellular Flux Analyzer". Sci Rep. 10(1):4239.
* For research use only.
