Methods for imaging labeled neurons together with neuropil features in Drosophila.
We describe staining protocols for serial semithin sections of Drosophila central ganglia that allow visualization of gene expression in particular neurons with counterstaining to display the ganglion architecture. Green fluorescent protein (GFP), expressed in a subset of sensory neurons from a selected enhancer trap line, is visualized by conventional immunohistochemistry with a peroxidase-linked antibody, and neural architecture is revealed by reduced silver staining. This makes visible in histological sections the same GFP-labeled cells seen with confocal microscopy, but with the especial advantage that neuropil structures are also revealed at the level of individual cells and neuron processes. Not only does this allow the physical relationships among intracellularly labeled neurons to be determined by reference to specific features in the neuropil but it also enables a function to be ascribed provisionally to particular regions of neuropil. These methods have particular utility for mapping morphological information on specific neurons in the context of central nervous system architecture, both in adult Drosophila and during development.
Tyrer
N M
NM
Department of Optometry and Neuroscience, University of Manchester Institute of Science and Technology, Manchester, United Kingdom. mark.tyrer@umist.ac.uk
Shepherd
D
D
Williams
D W
DW
D000818:Animals; D002460:Cell Line; D004331:Drosophila melanogaster / Q000378:metabolism*; D017952:Ganglia, Invertebrate / Q000166:cytology / Q000378:metabolism; D049452:Green Fluorescent Proteins; D007150:Immunohistochemistry; D008164:Luminescent Proteins / Q000378:metabolism*; D018613:Microscopy, Confocal; D009474:Neurons / Q000378:metabolism*; D019581:Neuropil / Q000378:metabolism*; D016622:Silver Staining
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins