<em>In vivo</em> Neuronal Calcium Imaging in <em>C. elegans</em>

Samuel H. Chung; Lin Sun; Christopher V. Gabel

doi:10.3791/50357

C. vivo Nöronal Kalsiyum Imaging'de elegans

JoVE Journal
Neuroscience

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JoVE Journal Neuroscience

In vivo Neuronal Calcium Imaging in C. elegans

C. vivo Nöronal Kalsiyum Imaging'de elegans

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11:06 min

April 10, 2013

DOI: 10.3791/50357-v

Samuel H. Chung*^1,2, Lin Sun*^1,2, Christopher V. Gabel^1,2

¹Department of Physiology and Biophysics,Boston University School of Medicine, ²Boston University Photonics Center

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Overview

This article discusses the use of genetically encoded fluorescent calcium indicators to measure calcium dynamics in single neurons of the nematode worm C. elegans. The technique allows for in vivo imaging of neuronal activity and cellular calcium physiology in response to sensory stimuli and cellular damage.

Key Study Components

Area of Science

Neuroscience
Cell Biology
Imaging Techniques

Background

C. elegans is a well-established model organism for neuronal studies.
It has a transparent body and well-documented neuroanatomy.
Genetic techniques allow for targeted expression of calcium indicators.
In vivo imaging provides insights into neuronal function and health.

Purpose of Study

To measure calcium dynamics in single neurons.
To analyze neuronal responses to sensory stimuli.
To investigate cellular responses to damage.

Methods Used

Mounting and demobilizing C. elegans expressing calcium-sensitive fluorophores.
In vivo imaging using video microscopy and epi-fluorescent techniques.
Analysis of fluorescence intensity at specific neuronal locations.
Comparison of two fluorophores: GCaMP and FRET-based Chameleon.

Main Results

Robust measurement of neuronal activity in response to stimuli.
Insights into calcium physiology during cellular damage.
Demonstration of the advantages of using C. elegans for imaging.
Technical simplicity and cost-effectiveness of the method.

Conclusions

C. elegans is a powerful model for in vivo neuronal imaging.
The technique is flexible and genetically tractable.
It provides valuable data on neuronal function and health.

Frequently Asked Questions

What is the main advantage of using C. elegans for neuronal imaging?

C. elegans offers a transparent body and well-documented neuroanatomy, making it ideal for in vivo imaging.

How are calcium dynamics measured in this study?

Calcium dynamics are measured using genetically encoded fluorescent indicators and video microscopy.

What types of fluorophores are used in the experiment?

The study uses GCaMP and FRET-based Chameleon fluorophores for imaging neuronal activity.

What are the applications of this imaging technique?

This technique can be used to study neuronal responses to sensory stimuli and cellular damage.

Is this method cost-effective?

Yes, the method is technically simple and cost-effective compared to other imaging techniques.

What insights can be gained from this study?

The study provides insights into neuronal function, calcium physiology, and the health of neurons in response to various stimuli.

Onun küçük şeffaf gövde, iyi belgelenmiş nöroanatomi ve müsait genetik teknikler ve reaktiflerin bir dizi ile,

Aşağıdaki deneyin genel amacı, nematod solucanı içindeki tek nöronların kalsiyum dinamiklerini ölçmek için genetik olarak kodlanmış floro dörtlüsünü kullanmaktır. Zarafete bakın, bu, ilk önce standart bir mikroskop lamı üzerinde ilgilenilen nöronlarda kalsiyuma duyarlı bir florofor ifade eden bozulmamış C zarafetinin monte edilmesi ve demobilize edilmesiyle elde edilir. Daha sonra, bireysel nöronlar, video mikroskobu ve standart epi floresan görüntüleme teknikleri kullanılarak in vivo olarak görüntülenir.

Daha sonra video, yaygın olarak kullanılan iki Fluor fours G kampı için nöronun belirli konumlarında floresan yoğunluğunu ölçmek için analiz edilir, bu da sağlam bir büyük genlikli tek kanallı sinyal ve çift kanal oranı oluşturan perde tabanlı bukalemun verir. Metrik ölçüm sonuçları, her ikisi de görüntülenen nöron içindeki bağıl kalsiyum seviyelerinin optik ölçümüne dayanan, belirli bir duyusal uyarana yanıt olarak gösterilen nöronal aktivitenin yanı sıra hücresel hasara yanıt olarak hücresel kalsiyum fizyolojisi elde edilir. Bu tekniğin omurgalı sistemlerindeki veya konvansiyonel elektrofizyolojideki benzer yöntemlere göre ana avantajları, CL elgan'ın teknik olarak basit ve uygun maliyetli olan in vivo nöronal görüntüleme için sağlam, esnek ve genetik olarak izlenebilir bir sistemi temsil etmesidir.

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