Gut Acidification Monitoring Assay: A Technique to Study the Acidification of the Intestinal Lumen in Drosophila using Bromophenol Blue Dye
Published: April 30, 2023
Abstract
Source: Abu, F., et al. Monitoring Gut Acidification in the Adult Drosophila Intestine. J. Vis. Exp. (2021).
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…
Divulgaciones
The authors have nothing to disclose.
Materials
| Bromophenol blue | Sigma-Aldrich | B0126 | |
| D. simulans | Drosophila Species Stock Center at the University of California | Riverside California1 (https://www.drosophilaspecies.com/) | |
| D. erecta | Drosophila Species Stock Center at the University of California | Dere cy1(https://www.drosophilaspecies.com/) | |
| D. pseudoobscura | Drosophila Species Stock Center at the University of California | Eugene, Oregon(https://www.drosophilaspecies.com/) | |
| D. mojavensis | Drosophila Species Stock Center at the University of California | Chocolate Mountains, California (https://www.drosophilaspecies.com/) | |
| Forceps | Inox Biology | Catalog# 11252-20 | |
| Kim wipes Tissue | Kimtech | ||
| Oregon R | Bloomington Drosophila Stock | (https://bdsc.indiana.edu/ # 2376) | |
| Petri dishes | Fisher Scientific | Catalog #FB0875713A | |
| Phosphate-buffered Saline (PBS) | HyClone | Catalog # SH30258.01 | |
| Stereomicroscope | Olympus SZ51 | Visual magnification |
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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).