Measuring Physiological Responses of <em>Drosophila</em> Sensory Neurons to Lipid Pheromones Using Live Calcium Imaging

Shruti Shankar; Meredith E.K. Calvert; Joanne Y. Yew

doi:10.3791/53392

Measuring Physiological Responses of Drosophila Sensory Neurons to Lipid Pheromones Usin...

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Immunology and Infection

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JoVE Journal Immunology and Infection

Measuring Physiological Responses of Drosophila Sensory Neurons to Lipid Pheromones Using Live Calcium Imaging

Measuring Physiological Responses of Drosophila Sensory Neurons to Lipid Pheromones Using Live Calcium Imaging

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07:25 min

April 29, 2016

DOI: 10.3791/53392-v

Shruti Shankar^1,2, Meredith E.K. Calvert^3,4, Joanne Y. Yew^1,2,5

¹Temasek Life Sciences Laboratory, ²Department of Biological Science,National University of Singapore, ³Bioimaging and Biocomputing Facility,Temasek Life Sciences Laboratory, ⁴Histology and Light Microscopy Core,Gladstone Institutes, ⁵Pacific Biosciences Research Center,University of Hawai'i at Mānoa

Overview

This study presents a method for measuring the physiological responses of gustatory sensory neurons in the forelegs and proboscis of Drosophila using calcium imaging. The technique allows for the examination of individual neuron responses in live flies to gustatory pheromones.

Key Study Components

Area of Science

Neuroscience
Behavioral Biology
Sensory Neurobiology

Background

Drosophila possess a variety of gustatory sensory neurons.
Understanding these neurons is crucial for sensory processing research.
Calcium imaging provides insights into neuronal activity.
Identifying ligands for orphan gustatory receptors is a key research goal.

Purpose of Study

To measure physiological responses of gustatory neurons to lipid pheromones.
To explore the functionality of gustatory receptors in live animals.
To advance knowledge in sensory neurobiology.

Methods Used

Calcium imaging technique applied to live Drosophila.
Anesthetizing the fly for stabilization during imaging.
Attaching the fly to a cover slip for observation.
Using a dissecting microscope to manipulate the fly's foreleg.

Main Results

Successful measurement of gustatory neuron responses to pheromones.
Individual neuron activity can be monitored in an intact organism.
Insights gained into the function of gustatory receptors.
Methodology validated for future sensory neurobiology studies.

Conclusions

The technique enhances understanding of gustatory neuron function.
It provides a framework for identifying receptor-ligand interactions.
Future studies can build on this methodology for deeper insights.

Frequently Asked Questions

What is the main focus of this study?

The study focuses on measuring the physiological responses of Drosophila gustatory neurons to lipid pheromones using calcium imaging.

Why is calcium imaging used in this research?

Calcium imaging allows for real-time observation of neuronal activity in live animals, providing insights into sensory processing.

What are orphan gustatory receptors?

Orphan gustatory receptors are receptors whose ligands have not yet been identified, making them a key target for research.

How are the flies prepared for imaging?

Flies are anesthetized and attached to a cover slip for stabilization during the imaging process.

What advantages does this method offer?

The method allows for the study of individual neuron responses in an intact organism, enhancing the understanding of sensory neurobiology.

The forelegs and proboscis of Drosophila contain a rich repertoire of gustatory sensory neurons. Here, we present a method using calcium imaging to measure physiological responses from sensory neurons in the foreleg and proboscis of live flies upon exogenous application of a gustatory pheromone.

The overall goal of this procedure is to measure the physiological responses of Drosophila gustatory neurons to lipid pheromones. This technique can be used to answer key questions in sensory neurobiology such as identifying the ligands for orphan gustatory receptors. The main advantage of this technique is that the responses of individual gustatory neurons can be studied in an intact, live animal.

After anesthetizing the fly, attach it to a 0.17 millimeter cover slip using a very small drop of clear nail polish. Attach the side of the fly to the slide using the entire drop. Next, under a dissecting microscope, use a wet paintbrush to extend a foreleg of the fly.

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