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Neuroscience
Isolamento de neurônios sensoriais de Aplysia californica Para Remendo Recordings Grampo...
Isolamento de neurônios sensoriais de Aplysia californica Para Remendo Recordings Grampo...
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Isolation of Sensory Neurons of Aplysia californica for Patch Clamp Recordings of Glutamatergic Currents

Isolamento de neurônios sensoriais de Aplysia californica Para Remendo Recordings Grampo de Correntes glutamatérgicos

Full Text
12,867 Views
09:40 min
July 10, 2013

DOI: 10.3791/50543-v

Lynne A. Fieber1, Stephen L. Carlson1, Andrew T. Kempsell1, Justin B. Greer1, Michael C. Schmale1

1Division of Marine Biology and Fisheries, Rosenstiel School of Marine and Atmospheric Sciences,University of Miami

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This article describes the dissection of the nervous system of the marine sea hare Aplysia for the isolation of sensory neurons. The procedure includes anesthesia, enzyme digestion, and patch clamp recordings to study ion currents.

Key Study Components

Area of Science

  • Neuroscience
  • Electrophysiology
  • Cell Culture

Background

  • The marine sea hare Aplysia is a model organism for studying neuronal function.
  • Isolation of sensory neurons allows for detailed electrophysiological studies.
  • Patch clamp technique is essential for measuring ion currents in neurons.
  • Short-term culture of neurons facilitates experimental manipulation.

Purpose of Study

  • To isolate individual sensory neurons for electrophysiological analysis.
  • To investigate voltage and agonist gated ion currents in these neurons.
  • To enhance understanding of neuronal signaling mechanisms.

Methods Used

  • Anesthesia of the marine sea hare Aplysia.
  • Dissection of ganglia containing sensory neurons.
  • Enzymatic digestion of ganglia overnight.
  • Dissociation and culture of sensory neurons for patch clamp studies.

Main Results

  • Successful isolation of sensory neurons from Aplysia.
  • Recording of ion currents using the patch clamp technique.
  • Characterization of voltage and agonist gated ion currents.
  • Insights into the electrophysiological properties of isolated neurons.

Conclusions

  • The procedure effectively isolates sensory neurons for study.
  • Patch clamp recordings provide valuable data on ion currents.
  • This method can be applied to further research in neuronal physiology.

Frequently Asked Questions

What is the significance of using Aplysia in neuroscience research?
Aplysia serves as a valuable model organism due to its large neurons and simple nervous system, making it ideal for studying fundamental neuronal processes.
How does the patch clamp technique work?
The patch clamp technique allows for the measurement of ionic currents through individual ion channels in neurons by creating a high-resistance seal with the cell membrane.
What are the advantages of short-term culture of neurons?
Short-term culture maintains the viability of neurons while allowing for experimental manipulation and observation of their physiological properties.
What types of ion currents can be studied using this method?
This method allows for the study of voltage-gated and agonist-gated ion currents, which are critical for understanding neuronal excitability and signaling.
Can this technique be applied to other species?
Yes, while this study focuses on Aplysia, similar techniques can be adapted for use in other species with suitable neuronal characteristics.

Nós descrevemos a dissecação do sistema nervoso do mar marinho lebre

O objetivo geral deste procedimento é isolar neurônios sensoriais individuais de plegia em cultura de curto prazo. Para estudos de patch clamp, isso é feito dissecando primeiro os gânglios contendo células sensoriais de um animal anestesiado. O segundo passo é digerir os gânglios em solução enzimática durante a noite.

Em seguida, os neurônios sensoriais de interesse são dissecados dos gânglios digeridos. A etapa final é a dissociação das placas de cultura preparadas para aderir ao substrato de cultura durante a noite. Em última análise, o clampeamento de tensão é usado para estudar a voltagem e as correntes de íons dependentes de agonistas expressas nesses neurônios.

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