Electrophysiological Recording from <em>Drosophila</em> Trichoid Sensilla in Response to Odorants of Low Volatility

Renny Ng; Hui-Hao Lin; Jing W. Wang; Chih-Ying Su

doi:10.3791/56147

Registrazione elettrofisiologica da Drosophila Senso Trichoide in risposta a Odoranti di...

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JoVE Journal Neuroscience

Electrophysiological Recording from Drosophila Trichoid Sensilla in Response to Odorants of Low Volatility

Registrazione elettrofisiologica da Drosophila Senso Trichoide in risposta a Odoranti di bassa volatilità

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07:49 min

July 27, 2017

DOI: 10.3791/56147-v

Renny Ng¹, Hui-Hao Lin¹, Jing W. Wang¹, Chih-Ying Su¹

¹Division of Biological Sciences,University of California San Diego

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Overview

This article presents a protocol for delivering low-volatility odorants to Drosophila olfactory receptor neurons during single-sensillum recordings. By using this method, researchers can investigate the response of these neurons to long-chain cuticular pheromones, enhancing our understanding of insect pheromone communication.

Key Study Components

Area of Science

Neuroscience
Electrophysiology
Insect Behavior

Background

Single-sensillum recordings allow for the study of individual olfactory receptor neurons.
Drosophila serves as a model organism to explore pheromone detection.
Low-volatility odorants pose challenges for effective presentation during recordings.
The developed technique aims to improve odorant delivery and neuronal response assessment.

Purpose of Study

To establish a reliable method for presenting low-volatility odorants to Drosophila olfactory neurons.
To facilitate the investigation of neuronal responses to specific pheromones.
To enhance knowledge of insect olfaction mechanisms.

Methods Used

Single-sensillum recordings were conducted on Drosophila olfactory receptor neurons.
The protocol included detailed steps for preparing odor delivery cartridges and fly preparations.
Electrophysiological recordings focused on the response of neurons to different concentrations of palmitoleic acid.
Emphasis was placed on careful positioning of the odor cartridge relative to the fly's antenna.
Recordings were taken at varying distances to assess effective stimulus delivery.

Main Results

Consistency in cartridge positioning significantly affected neuronal response rates.
Effective presentation of palmitoleic acid from 4mm distance led to increased neuronal spikes, while 11mm distance resulted in minimal responses.
The technique demonstrated that trans-palmitoleic acid was a stronger stimulant than its cis form for Or47b neurons.

Conclusions

This study establishes an effective method for presenting low-volatility pheromones to olfactory neurons.
The findings advance the understanding of mechanisms involved in the detection of pheromones in insects.
By mastering this technique, researchers can explore underlying neurophysiological processes in olfaction.

Frequently Asked Questions

What is the advantage of using Drosophila for sensory studies?

Drosophila is a powerful model organism for studying olfactory systems due to its well-mapped genome and defined neuronal pathways, making it easier to understand nervous system function.

How is the odorant presentation implemented?

Odorants are delivered via specialized cartridges positioned closely to the Drosophila antenna, allowing for the effective stimulation of olfactory neurons.

What outcomes can be measured using this method?

The method allows for the measurement of neuronal responses in terms of spike rates and can highlight differences in response to various pheromone concentrations.

How can this method be adapted for other low-volatility compounds?

The cartridge preparation procedure can be modified to suit different low-volatility odorants, allowing diverse chemical studies in Drosophila olfaction.

What are key limitations to consider with this technique?

Precision in positioning the odorant cartridge is critical; inaccuracies can lead to inconsistent results in neuronal spike response, which may affect data reliability.

L'obiettivo generale di questo protocollo è quello di dimostrare come presentare gli odoranti a bassa volatilità per la registrazione singolo-sensillare da parte dei neuroni del recettore olfattivo Drosophila che rispondono ai feromoni cuticolari a catena lunga.

L'obiettivo generale di questa tecnica è quello di presentare efficacemente odoranti a bassa volatilità ai neuroni dei recettori olfattivi e alla drosofila per la registrazione elettrofisiologica. Questi neuroni sono alloggiati nei sensilli tricoidi e rispondono ai feromoni cuticolari a catena lunga. Questo metodo può aiutare a rispondere a domande chiave nel campo della comunicazione dei feromoni degli insetti fornendo una migliore tecnica di erogazione degli odori.

Il vantaggio principale di questa tecnica è che consente agli odorizzanti che in precedenza erano inaccessibili a causa della loro bassa volatilità di essere presentati efficacemente durante la registrazione in vivo. In particolare, i feromoni vengono presentati da distanze ravvicinate che simulano la distanza tra i maschi che corteggiano e le femmine bersaglio. La preparazione del componente della cartuccia di erogazione degli odori per le registrazioni AT4 inizia con la rimozione di 0,9 centimetri dall'estremità stretta di un puntale per pipetta da 200 microlitri.

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