A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains.

J Vis Exp 2016 12 1(118). Epub 2016 Dec 1.

Center for Metabolic and Degenerative Diseases, The Brown Foundation Institute of Molecular Medicine, Programs in Human and Molecular Genetics and Neuroscience, The Graduate School of Biomedical Sciences at Houston, The University of Texas Health Science Center at Houston; Department of Neurobiology and Anatomy, McGovern Medical School, The University of Texas Health Science Center at Houston;

There is an increasing interest in using Drosophila to model human brain degenerative diseases, map neuronal circuitries in adult brains, and study the molecular and cellular basis of higher brain functions. A whole-mount preparation of adult brains with well-preserved morphology is critical for such whole brain-based studies, but can be technically challenging and time-consuming. This protocol describes an easy-to-learn, one-step dissection approach of an adult fly head in less than 10 s, while keeping the intact brain attached to the rest of the body to facilitate subsequent processing steps. The procedure helps remove most of the eye and tracheal tissues normally associated with the brain that can interfere with the later imaging step, and also places less demand on the quality of the dissecting forceps. Additionally, we describe a simple method that allows convenient flipping of the mounted brain samples on a coverslip, which is important for imaging both sides of the brains with similar signal intensity and quality. As an example of the protocol, we present an analysis of dopaminergic (DA) neurons in adult brains of WT (w) flies. The high efficacy of the dissection method makes it particularly useful for large-scale adult brain-based studies in Drosophila.

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Source
http://dx.doi.org/10.3791/55128DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5226341PMC
December 2016

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