Literature DB >> 21386006

Optogenetics in the teaching laboratory: using channelrhodopsin-2 to study the neural basis of behavior and synaptic physiology in Drosophila.

Stefan R Pulver1, Nicholas J Hornstein, Bruce L Land, Bruce R Johnson.   

Abstract

Here we incorporate recent advances in Drosophila neurogenetics and "optogenetics" into neuroscience laboratory exercises. We used the light-activated ion channel channelrhodopsin-2 (ChR2) and tissue-specific genetic expression techniques to study the neural basis of behavior in Drosophila larvae. We designed and implemented exercises using inexpensive, easy-to-use systems for delivering blue light pulses with fine temporal control. Students first examined the behavioral effects of activating glutamatergic neurons in Drosophila larvae and then recorded excitatory junctional potentials (EJPs) mediated by ChR2 activation at the larval neuromuscular junction (NMJ). Comparison of electrically and light-evoked EJPs demonstrates that the amplitudes and time courses of light-evoked EJPs are not significantly different from those generated by electrical nerve stimulation. These exercises introduce students to new genetic technology for remotely manipulating neural activity, and they simplify the process of recording EJPs at the Drosophila larval NMJ. Relatively little research work has been done using ChR2 in Drosophila, so students have opportunities to test novel hypotheses and make tangible contributions to the scientific record. Qualitative and quantitative assessment of student experiences suggest that these exercises help convey principles of synaptic transmission while also promoting integrative and inquiry-based studies of genetics, cellular physiology, and animal behavior.

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Year:  2011        PMID: 21386006      PMCID: PMC3276384          DOI: 10.1152/advan.00125.2010

Source DB:  PubMed          Journal:  Adv Physiol Educ        ISSN: 1043-4046            Impact factor:   2.288


  30 in total

1.  Drosophila cholinergic neurons and processes visualized with Gal4/UAS-GFP.

Authors:  P M Salvaterra; T Kitamoto
Journal:  Brain Res Gene Expr Patterns       Date:  2001-08

2.  Light activation of channelrhodopsin-2 in excitable cells of Caenorhabditis elegans triggers rapid behavioral responses.

Authors:  Georg Nagel; Martin Brauner; Jana F Liewald; Nona Adeishvili; Ernst Bamberg; Alexander Gottschalk
Journal:  Curr Biol       Date:  2005-12-20       Impact factor: 10.834

3.  Millisecond-timescale, genetically targeted optical control of neural activity.

Authors:  Edward S Boyden; Feng Zhang; Ernst Bamberg; Georg Nagel; Karl Deisseroth
Journal:  Nat Neurosci       Date:  2005-08-14       Impact factor: 24.884

4.  Properties of the larval neuromuscular junction in Drosophila melanogaster.

Authors:  L Y Jan; Y N Jan
Journal:  J Physiol       Date:  1976-10       Impact factor: 5.182

5.  A comparison of electrically evoked and channel rhodopsin-evoked postsynaptic potentials in the pharyngeal system of Caenorhabditis elegans.

Authors:  Christopher J Franks; Caitriona Murray; David Ogden; Vincent O'Connor; Lindy Holden-Dye
Journal:  Invert Neurosci       Date:  2009-03-18

6.  A toolbox for light control of Drosophila behaviors through Channelrhodopsin 2-mediated photoactivation of targeted neurons.

Authors:  Wei Zhang; Wooping Ge; Zuoren Wang
Journal:  Eur J Neurosci       Date:  2007-10-26       Impact factor: 3.386

7.  Channelrhodopsin-2, a directly light-gated cation-selective membrane channel.

Authors:  Georg Nagel; Tanjef Szellas; Wolfram Huhn; Suneel Kateriya; Nona Adeishvili; Peter Berthold; Doris Ollig; Peter Hegemann; Ernst Bamberg
Journal:  Proc Natl Acad Sci U S A       Date:  2003-11-13       Impact factor: 11.205

8.  Using Neurogenetics and the Warmth-Gated Ion Channel TRPA1 to Study the Neural Basis of Behavior in Drosophila.

Authors:  Jimena Berni; Alistair M Muldal; Stefan R Pulver
Journal:  J Undergrad Neurosci Educ       Date:  2010-10-15

9.  g-PRIME: A Free, Windows Based Data Acquisition and Event Analysis Software Package for Physiology in Classrooms and Research Labs.

Authors:  Gus K Lott; Bruce R Johnson; Robert H Bonow; Bruce R Land; Ronald R Hoy
Journal:  J Undergrad Neurosci Educ       Date:  2009-10-15

10.  Targeted gene expression as a means of altering cell fates and generating dominant phenotypes.

Authors:  A H Brand; N Perrimon
Journal:  Development       Date:  1993-06       Impact factor: 6.868

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  20 in total

1.  Investigation of Seizure-Susceptibility in a Drosophila melanogaster Model of Human Epilepsy with Optogenetic Stimulation.

Authors:  Arunesh Saras; Veronica V Wu; Harlan J Brawer; Mark A Tanouye
Journal:  Genetics       Date:  2017-06-19       Impact factor: 4.562

2.  Recording field potentials from zebrafish larvae during escape responses.

Authors:  Bryan D Monesson-Olson; Eileen L Troconis; Josef G Trapani
Journal:  J Undergrad Neurosci Educ       Date:  2014-10-15

3.  Grasshopper DCMD: An Undergraduate Electrophysiology Lab for Investigating Single-Unit Responses to Behaviorally-Relevant Stimuli.

Authors:  Dieu My T Nguyen; Mark Roper; Stanislav Mircic; Robert M Olberg; Gregory J Gage
Journal:  J Undergrad Neurosci Educ       Date:  2017-06-15

4.  Reducing the Cost of Electrophysiology in the Teaching Laboratory.

Authors:  Robert A Wyttenbach; Bruce R Johnson; Ronald R Hoy
Journal:  J Undergrad Neurosci Educ       Date:  2018-09-15

5.  Aversive and Appetitive Learning in Drosophila Larvae: A Simple and Powerful Suite of Laboratory Modules for Classroom or Open-ended Research Projects.

Authors:  Austin Pavin; Kevin Fain; Allison DeHart; Divya Sitaraman
Journal:  J Undergrad Neurosci Educ       Date:  2018-06-15

6.  Portable conduction velocity experiments using earthworms for the college and high school neuroscience teaching laboratory.

Authors:  Kyle M Shannon; Gregory J Gage; Aleksandra Jankovic; W Jeffrey Wilson; Timothy C Marzullo
Journal:  Adv Physiol Educ       Date:  2014-03       Impact factor: 2.288

7.  A Protocol Demonstrating 60 Different Drosophila Behaviors in One Assay.

Authors:  Claire E McKellar; Robert A Wyttenbach
Journal:  J Undergrad Neurosci Educ       Date:  2017-06-15

8.  Using Optogenetics to Understand Neuronal Mechanisms Underlying Behavior in C. elegans.

Authors:  Navin Pokala; Elizabeth E Glater
Journal:  J Undergrad Neurosci Educ       Date:  2018-06-15

9.  Probing Synaptic Transmission and Behavior in Drosophila with Optogenetics: A Laboratory Exercise.

Authors:  Ilya Vilinsky; Karen L Hibbard; Bruce R Johnson; David L Deitcher
Journal:  J Undergrad Neurosci Educ       Date:  2018-09-15

10.  Light Activated Escape Circuits: A Behavior and Neurophysiology Lab Module using Drosophila Optogenetics.

Authors:  Josh S Titlow; Bruce R Johnson; Stefan R Pulver
Journal:  J Undergrad Neurosci Educ       Date:  2015-07-07
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