Acquisition of High-Quality Digital Video of Drosophila Larval and Adult Behaviors from a Lateral Perspective

Summary

Here we describe a simple and widely accessible microscopy technique to acquire high-quality digital video of Drosophila adult and larval mutant phenotypes from a lateral perspective.

Abstract

Drosophila melanogaster is a powerful experimental model system for studying the function of the nervous system. Gene mutations that cause dysfunction of the nervous system often produce viable larvae and adults that have locomotion defective phenotypes that are difficult to adequately describe with text or completely represent with a single photographic image. Current modes of scientific publishing, however, support the submission of digital video media as supplemental material to accompany a manuscript. Here we describe a simple and widely accessible microscopy technique for acquiring high-quality digital video of both Drosophila larval and adult phenotypes from a lateral perspective. Video of larval and adult locomotion from a side-view is advantageous because it allows the observation and analysis of subtle distinctions and variations in aberrant locomotive behaviors. We have successfully used the technique to visualize and quantify aberrant crawling behaviors in third instar larvae, in addition to adult mutant phenotypes and behaviors including grooming.

Introduction

The common fruit fly Drosophila melanogaster is a powerful experimental model system for studying the function of the nervous system^1-3. Evolutionary conservation of structure and function of the nervous system with humans, as well as ease of genetic manipulation and a vast array of genetic tools makes Drosophila the premiere organism to model human neurodegenerative diseases⁴. Gene mutations that cause dysfunction of the nervous system often result in viable mutant larvae and adult Drosophila with impaired locomotion. Phenotypes observed in nervous system defective mutants include reduced rate of locomotion, a....

Protocol

1. The Stereo Microscope System

Note: Although this protocol is easily adaptable to virtually any stereo microscope system coupled to a digital camera with the capability of acquiring video, details are provided on the system used in our lab (Table of Materials/Equipment).

1. Acquire digital video using a trinocular stereo microscope coupled to a commercial digital camera.
2. In order to couple the commercial digital camera to the trinocular port of the stereo microscope, remove the ½x C-mount of the phototube port of the stereo microscope and replace it with a 1X C-mount.
3. Mount a digital camera coupler (....

Results

We have successfully used this technique to acquire and quantify the larval behavioral phenotype associated with loss of function of the stathmin gene (Figure 2)¹⁴. The stathmin gene encodes a microtubule regulatory protein that partitions tubulin dimers from pools of soluble tubulin, and binds microtubules and promotes their disassembly^15,16. Stathmin function is required to maintain the integrity of microtubules in the axons of peripheral nerves¹⁴. Dis.......

Discussion

Drosophila melanogaster’s strength as a model system for studying nervous system function stems largely from the convergence of the powerful genetic tools available and the broad array of robust behavioral assays developed. Here we present a simple and widely accessible microscopy technique for acquiring high-quality digital video of Drosophila adult and larval locomotive phenotypes from a lateral perspective. We have successfully used this approach to characterize and quantify the severity of pos.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Trinocular Stereozoom Microscope	Olympus Corporation	SZ6145TR	½ C-mount was removed and replaced with 1X C-mount
1X C-mount	Leeds Precision Instruments	LSZ-1XCMT2
Digital Camera Coupler (43 mm thread)	Qioptiq Imaging Solutions	25-70-10-02
58 mm to 48 mm Step Down Ring	B&H Video	GBSDR5848
48 mm to 43 mm Step Down Ring	B&H Video	GBSDR4843
Lensmate Adapter Kit for Canon G10	LensMateOnline.com
Canon PowerShot G10 Digital Camera	Canon U.S.A., Inc.
1.5 ml Spectroscopic Polysterene Cuvette	Denville Scientific	U8650-4