C. elegans Membrane Fluidity Assessment Service

Membrane homeostasis is a defining feature of all cells, and biological membranes are primarily composed of phospholipids. The properties of phospholipid bilayer are greatly influenced by the proportion of two fatty acid species (saturated fatty acid and unsaturated fatty acid). A high unsaturated fatty acid content promotes membrane fluidity, while a high saturated fatty acid content increases membrane rigidity. The membrane composition adaptation involved adjustment strategies to deal with environmental conditions such as pH, temperature, and pressure almost in all of life’s diversity. It is reported that the specific composition of subcellular membranes plays vital roles in many cellular processes, such as vesicular trafficking, receptor signaling and organelle homeostasis. Recent studies indicate the importance of specific polyunsaturated lipids in the process of membrane deformation, domain stability and even in membrane fission events. Therefore, it is not surprising to find membrane composition defects are connected with many diseases, including cancers, diabetics, etc. For example, the membrane composition of cells is correlated with transformation in breast cancer. And for diabetics, the rigid cellular membranes are rich in saturated fatty acids, which may be one of the useful diagnostic criteria for the diseases.

The membrane homeostasis in C. elegans

For animals, there is a big challenge to maintain membrane homeostasis due to the structural fatty acids (FAs) of membrane being primarily acquired from a highly variable diet. For C. elegans, the nematode daily uses mostly dietary fatty acids as substitutive building blocks of nearly 80% of its membrane phospholipids. Since the high variation in dietary composition and narrow membrane composition range for maintenance of membrane homeostasis, it is obvious that there are robust regulatory mechanisms adjusting FAs composition and compensating for dietary variation. Previous studies have demonstrated that the proteins PAQR-2 and IGLR-2 exist in C. elegans. The proteins act as sensors in the plasma membrane to facilitate fatty acid desaturation and recovery of liquidity in specific conditions. Particularly, the C. elegans PAQR-2 is the homolog of human AdipoR1/2, which also exhibits function to regulate membrane fluidity in human cells.

Evaluation of membrane fluidity service in Creative Biogene

The new or improved methods, including lipidomic analysis of membrane composition, fluorescence recovery after photobleaching (FRAP), molecular modelling of membranes and powerful genetic approaches have recently been exploited from different perspectives to identify the regulators and mechanisms of membrane homeostasis. Among them, FRAP is probably the most direct method to assess membrane fluidity in vivo. Herein, we offer FRAP to investigate the dynamics of molecules in C. elegans. FRAP is conducted with confocal laser scanning microscopes. Fluorescent molecules in a specified region within the plasma membrane are photobleached by a high-intensity laser. Then the resulting recovery of fluorescent intensity of bleached regions is monitored and plotted on a recovery curve, and finally quantified. Additionally, a green fluorescent protein (GFP) is applied in FRAP, a most common way to fluorescently tag proteins in living cells.

Workflow of photobleaching experiments

Workflow of photobleaching experiments - Creative Biogene

Creative Biogene offers FRAP method to investigate the dynamics of molecules in C. elegans. We are dedicated to assisting our customers to explore the mechanisms of membrane fluidity in C. elegans and even in human cells and related diseases. If you are interested in this area, please contact us. We are glad to cooperate with you.

Reference

  1. Devkota, R. and Pilon, M. (2018). "FRAP: A Powerful Method to Evaluate Membrane Fluidity in Caenorhabditis elegans". Bio-protocol. 8(13): e2913.

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