An Affordable and Efficient ``Homemade{''} Platform for Drosophila Behavioral Studies, and an Accompanying Protocol for Larval Mitochondrial Respirometry

Author(s)

Garcia, Geovana S. ^[1] ; Othonicar, Murilo F. ^[1] ; Oliveira, Marcos T. ^[1] ; Couto-Lima, Carlos A. ^[2]

Affiliation

^[1] Univ Estadual Paulista, Fac Ciencias Agr & Veterinarias, Dept Tecnol, Presidente Prudente - Brazil

^[2] Univ Oeste Paulista, Fac Ciencias Saude, Biomed, Presidente Prudente - Brazil

• Document type: Journal article
• Source: JOVE-JOURNAL OF VISUALIZED EXPERIMENTS; n. 175 SEP 2021.
• Web of Science Citations: 0

Abstract

The usefulness of Drosophila as a model organism for the study of human diseases, behaviors and basic biology is unquestionable. Although practical, Drosophila research lacks popularity in developing countries, possibly due to the misinformed idea that establishing a lab and performing relevant experiments with such tiny insects is difficult and requires expensive, specialized apparatuses. Here, we describe how to build an affordable flylab to quantitatively analyze a myriad of behavioral parameters in D. melanogaster, by 3D-printing many of the necessary pieces of equipment. We provide protocols to build in-house vial racks, courtship arenas, apparatuses for locomotor assays, etc., to be used for general fly maintenance and to perform behavioral experiments using adult flies and larvae. We also provide protocols on how to use more sophisticated systems, such as a high resolution oxygraph, to measure mitochondrial oxygen consumption in larval samples, and show its association with behavioral changes in the larvae upon the xenotopic expression of the mitochondrial alternative oxidase (AOX). AOX increases larval activity and mitochondrial leak respiration, and accelerates development at low temperatures, which is consistent with a thermogenic role for the enzyme. We hope these protocols will inspire researchers, especially from developing countries, to use Drosophila to easily combine behavior and mitochondrial metabolism data, which may lead to information on genes and/ or environmental conditions that may also regulate human physiology and disease states. (AU)

• FAPESP's process: 14/02253-6 - Investigating the metabolic alterations caused by the transgenic expression of the mitochondrial alternative oxidase of Ciona intestinalis in Drosophila melanogaster
• Grantee: Marcos Túlio de Oliveira
• Support Opportunities: Research Grants - Young Investigators Grants
• FAPESP's process: 17/04372-0 - Mitochondrial DNA: mechanisms for genome integrity maintenance and impact on disease
• Grantee: Nadja Cristhina de Souza Pinto
• Support Opportunities: Research Projects - Thematic Grants