Abstract
The purpose of this experiment was to determine the effect of fish oil containing omega-3 fatty acids on distribution of proteins flotillin and caveolin associated with lipid microdomains in bovine luteal cells. Lipid microdomains are small, organized areas of the cell membrane composed of cholesterol, sphingolipids, and proteins. Lipid microdomains are believed to be connected to cell signaling. Adding fish oil containing omega-3 fatty acids is believed to disrupt cell signaling that occurs within lipid microdomains. Mixed luteal cells were prepared from four bovine corpora lutea. Tissue was digested using collagenase-1 and incubated in T-75 flasks containing Ham's F-12 medium with 5% fetal bovine serum, insulin, transferrin (5µg/ml), selenium (5ng/ml), 100 U/ml penicillin, 0.1mg/ml streptomycin, and 0.25 mg/ml amphotericin B in an atmosphere of humidified 95% air and 5% CO2 at 37ºC. At confluence, cells either remained untreated, control, or treated with 0.03% fish oil for a period of 48 hours. 10mM of Beta methyl cyclodextrin (βMCD), which is a reagent known to remove cholesterol, was used as a negative control. Cells underwent protein isolation and western blot imaging and analysis. Control didn't have a dispersion of proteins associated with lipid microdomains into the bulk lipids, while both βMCD and fish oil treated cells demonstrated a shift in proteins from the lipid microdomains into bulk lipids. In conclusion, the addition of fish oil appears to have disrupted the cholesterol, sphingolipids, and proteins found in lipid microdomains and could have affected signal transduction within bovine
luteal cells.
luteal cells.