This video demonstrates an assay to monitor gut acidification in Drosophila flies. After the ingestion of food supplemented with a pH indicator dye, the acidification of the intestinal lumen results in a change in the color of the dye. Visual inspection of the digestive tract of the flies to examine the color change provides evidence of acidification.
Protocol
NOTE: The standard laboratory line Oregon R was used as a WT control. All flies were reared on standard cornmeal-molasses medium (containing molasses, agar, yeast, cornmeal, tegosept, propionic acid, and water) at room temperature with 12/12 h light/dark circadian rhythm. 1. Preparing for the assay Collect female flies (0-2 days old, non-virgin) under CO2 anesthesia and allow them to recover on standard cornmeal food for at least 3 days before exper…
Representative Results
Figure 1: Gut acidification monitoring. (A) Schematic drawing of feeding arena. The blue dot represents fly food with bromophenol blue (a pH-indicating dye). Other spots represent fruit flies. (B) Graphical representation of percentage of flies showing gut acidification fed for different durations over 4 h. Representative gut images of an acidified gut and a…
Divulgations
The authors have nothing to disclose.
Materials
Bromophenol blue
Sigma-Aldrich
B0126
D. simulans
Drosophila Species Stock Center at the University of California
Gut Acidification Monitoring Assay: A Technique to Study the Acidification of the Intestinal Lumen in Drosophila using Bromophenol Blue Dye. J. Vis. Exp. (Pending Publication), e21239, doi: (2023).