Oxygen is essential for metazoans to survive. During natural development and homeostasis, cells and tissues are often challenged by a low level of oxygen condition, which is termed hypoxia for normal physiological functions. Hypoxia can lead to decreased metabolic rate, increased glycolysis, and pausing or slowing of the cell cycle. Apart from being affected by normal development or changes in environmental conditions, oxygen levels are severely affected by disease states such as cancer and various heart and lung diseases. Understanding the cellular signaling pathways that are associated with the response to hypoxia could provide new insight into the various strategies dealing with these pathologies in humans.
The C. elegans do not have specific respiratory organs, the cavity of which is filled with fluid allowing rapid exchange of gases across the cells, as opposed to a facilitated respiratory system. However, due to the unique characteristics, it is possible to study cellular responses to specific hypoxic O2 concentrations without confounding effects. Moreover, factors known to be referred to as the response to hypoxia are conserved in C. elegans, including the highly-conserved hypoxia inducible factor-1 (HIF-1, the ortholog of HIF in humans). In addition, it is easy to culture and genetically manipulate. The C. elegans are ideally suited as a model organism for the study of hypoxic response.
In normal cultural conditions, C. elegans can survive in anoxia for about a day, by entering a reversible state known as the suspended animation. As for CO2 fluctuation, previous studies reveal that the worms show a wide CO2 tolerance compared to other animals. Other studies have shown that C. elegans avoids high levels of CO2, and the intensity is suppressed under starvation conditions. Furthermore, the sensitivity of O2 and CO2 is closely related to food-sensing pathways, especially the ILS pathway, involving a transcription factor DAF-16 (homolog to mammalian FOXO). The hypoxic environment activates hypoxia-inducible factor-1 (hif-1) in C. elegans, which is similar to humans. This activation plays a central role in tissue repair, ischemia, and cancer. In normal O2 concentrations, the protein HIF-1 is degraded by a prolyl-hydroxylase encoded by egl-9, which is the ortholog EGLN/PHD in mammals.
Visualization of C. elegans in hypoxia with microscopy
(Fawcett EM, et al, 2012)The fact that different physiological responses occur in different hypoxic conditions highlights the importance of advanced experiment equipment and an optimal method to control the experimental conditions. We offer reliable and reproducible hypoxic conditions with defined concentrations of O2 within operational equipment and a practical method. The constructional environmental chambers are transparent and small, which can be placed on the dissecting scope, allowing an easily maneuverable for optimal visualization of worms in hypoxia. Besides, many hypoxia-induced effects are quickly reversed upon returning to room air. Rapid harvest of hypoxia-exposed worms can be achieved on this platform. Furthermore, we conduct the continual flow method that ensures rapid equilibration of the chamber and increases the stability of the system.
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