Cargando…

Simultaneous Whole-cell Recordings from Photoreceptors and Second-order Neurons in an Amphibian Retinal Slice Preparation

One of the central tasks in retinal neuroscience is to understand the circuitry of retinal neurons and how those connections are responsible for shaping the signals transmitted to the brain. Photons are detected in the retina by rod and cone photoreceptors, which convert that energy into an electric...

Descripción completa

Detalles Bibliográficos
Autores principales: Van Hook, Matthew J., Thoreson, Wallace B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MyJove Corporation 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3724563/
https://www.ncbi.nlm.nih.gov/pubmed/23770753
http://dx.doi.org/10.3791/50007
_version_ 1782476694960996352
author Van Hook, Matthew J.
Thoreson, Wallace B.
author_facet Van Hook, Matthew J.
Thoreson, Wallace B.
author_sort Van Hook, Matthew J.
collection PubMed
description One of the central tasks in retinal neuroscience is to understand the circuitry of retinal neurons and how those connections are responsible for shaping the signals transmitted to the brain. Photons are detected in the retina by rod and cone photoreceptors, which convert that energy into an electrical signal, transmitting it to other retinal neurons, where it is processed and communicated to central targets in the brain via the optic nerve. Important early insights into retinal circuitry and visual processing came from the histological studies of Cajal(1,2) and, later, from electrophysiological recordings of the spiking activity of retinal ganglion cells - the output cells of the retina(3,4). A detailed understanding of visual processing in the retina requires an understanding of the signaling at each step in the pathway from photoreceptor to retinal ganglion cell. However, many retinal cell types are buried deep in the tissue and therefore relatively inaccessible for electrophysiological recording. This limitation can be overcome by working with vertical slices, in which cells residing within each of the retinal layers are clearly visible and accessible for electrophysiological recording. Here, we describe a method for making vertical sections of retinas from larval tiger salamanders (Ambystoma tigrinum). While this preparation was originally developed for recordings with sharp microelectrodes(5,6), we describe a method for dual whole-cell voltage clamp recordings from photoreceptors and second-order horizontal and bipolar cells in which we manipulate the photoreceptor's membrane potential while simultaneously recording post-synaptic responses in horizontal or bipolar cells. The photoreceptors of the tiger salamander are considerably larger than those of mammalian species, making this an ideal preparation in which to undertake this technically challenging experimental approach. These experiments are described with an eye toward probing the signaling properties of the synaptic ribbon - a specialized synaptic structure found in a only a handful of neurons, including rod and cone photoreceptors, that is well suited for maintaining a high rate of tonic neurotransmitter release(7,8) - and how it contributes to the unique signaling properties of this first retinal synapse.
format Online
Article
Text
id pubmed-3724563
institution National Center for Biotechnology Information
language English
publishDate 2013
publisher MyJove Corporation
record_format MEDLINE/PubMed
spelling pubmed-37245632013-07-30 Simultaneous Whole-cell Recordings from Photoreceptors and Second-order Neurons in an Amphibian Retinal Slice Preparation Van Hook, Matthew J. Thoreson, Wallace B. J Vis Exp Neuroscience One of the central tasks in retinal neuroscience is to understand the circuitry of retinal neurons and how those connections are responsible for shaping the signals transmitted to the brain. Photons are detected in the retina by rod and cone photoreceptors, which convert that energy into an electrical signal, transmitting it to other retinal neurons, where it is processed and communicated to central targets in the brain via the optic nerve. Important early insights into retinal circuitry and visual processing came from the histological studies of Cajal(1,2) and, later, from electrophysiological recordings of the spiking activity of retinal ganglion cells - the output cells of the retina(3,4). A detailed understanding of visual processing in the retina requires an understanding of the signaling at each step in the pathway from photoreceptor to retinal ganglion cell. However, many retinal cell types are buried deep in the tissue and therefore relatively inaccessible for electrophysiological recording. This limitation can be overcome by working with vertical slices, in which cells residing within each of the retinal layers are clearly visible and accessible for electrophysiological recording. Here, we describe a method for making vertical sections of retinas from larval tiger salamanders (Ambystoma tigrinum). While this preparation was originally developed for recordings with sharp microelectrodes(5,6), we describe a method for dual whole-cell voltage clamp recordings from photoreceptors and second-order horizontal and bipolar cells in which we manipulate the photoreceptor's membrane potential while simultaneously recording post-synaptic responses in horizontal or bipolar cells. The photoreceptors of the tiger salamander are considerably larger than those of mammalian species, making this an ideal preparation in which to undertake this technically challenging experimental approach. These experiments are described with an eye toward probing the signaling properties of the synaptic ribbon - a specialized synaptic structure found in a only a handful of neurons, including rod and cone photoreceptors, that is well suited for maintaining a high rate of tonic neurotransmitter release(7,8) - and how it contributes to the unique signaling properties of this first retinal synapse. MyJove Corporation 2013-06-01 /pmc/articles/PMC3724563/ /pubmed/23770753 http://dx.doi.org/10.3791/50007 Text en Copyright © 2013, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Neuroscience
Van Hook, Matthew J.
Thoreson, Wallace B.
Simultaneous Whole-cell Recordings from Photoreceptors and Second-order Neurons in an Amphibian Retinal Slice Preparation
title Simultaneous Whole-cell Recordings from Photoreceptors and Second-order Neurons in an Amphibian Retinal Slice Preparation
title_full Simultaneous Whole-cell Recordings from Photoreceptors and Second-order Neurons in an Amphibian Retinal Slice Preparation
title_fullStr Simultaneous Whole-cell Recordings from Photoreceptors and Second-order Neurons in an Amphibian Retinal Slice Preparation
title_full_unstemmed Simultaneous Whole-cell Recordings from Photoreceptors and Second-order Neurons in an Amphibian Retinal Slice Preparation
title_short Simultaneous Whole-cell Recordings from Photoreceptors and Second-order Neurons in an Amphibian Retinal Slice Preparation
title_sort simultaneous whole-cell recordings from photoreceptors and second-order neurons in an amphibian retinal slice preparation
topic Neuroscience
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3724563/
https://www.ncbi.nlm.nih.gov/pubmed/23770753
http://dx.doi.org/10.3791/50007
work_keys_str_mv AT vanhookmatthewj simultaneouswholecellrecordingsfromphotoreceptorsandsecondorderneuronsinanamphibianretinalslicepreparation
AT thoresonwallaceb simultaneouswholecellrecordingsfromphotoreceptorsandsecondorderneuronsinanamphibianretinalslicepreparation