Juusola, M. orcid.org/0000-0002-4428-5330, Dau, A., Zheng, L. et al. (1 more author) (2016) Electrophysiological method for recording intracellular voltage responses of Drosophila photoreceptors and interneurons to light stimuli in vivo. Journal of Visualized Experiments (112). e54142. ISSN 1940-087X
Abstract
Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method described here enables the investigator to measure long-lasting (from minutes to hours) high-quality intracellular responses from single Drosophila R1-R6 photoreceptors and Large Monopolar Cells (LMCs) to light stimuli. Because the recording system has low noise, it can be used to study variability among individual cells in the fly eye, and how their outputs reflect the physical properties of the visual environment. We outline all key steps in performing this technique. The basic steps in constructing an appropriate electrophysiology set-up for recording, such as design and selection of the experimental equipment are described. We also explain how to prepare for recording by making appropriate (sharp) recording and (blunt) reference electrodes. Details are given on how to fix an intact fly in a bespoke fly-holder, prepare a small window in its eye and insert a recording electrode through this hole with minimal damage. We explain how to localize the center of a cell's receptive field, dark- or light-adapt the studied cell, and to record its voltage responses to dynamic light stimuli. Finally, we describe the criteria for stable normal recordings, show characteristic high-quality voltage responses of individual cells to different light stimuli, and briefly define how to quantify their signaling performance. Many aspects of the method are technically challenging and require practice and patience to master. But once learned and optimized for the investigator's experimental objectives, it grants outstanding in vivo neurophysiological data
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2016 Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License |
Keywords: | Neuroscience; Issue 112; Sharp microelectrode; current-clamp; electrophysiology; signaling performance; neural response; insect vision; signal-to-noise ratio; neuroethology; naturalistic light stimulation |
Dates: |
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Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Science (Sheffield) > School of Biosciences (Sheffield) |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 09 Aug 2023 16:00 |
Last Modified: | 10 Aug 2023 13:01 |
Status: | Published |
Publisher: | MyJove Corporation |
Refereed: | Yes |
Identification Number: | 10.3791/54142-v |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:202271 |