scholarly journals Patch Clamp Recordings from Mouse Retinal Neurons in a Dark-adapted Slice Preparation

10.3791/2107 ◽  
2010 ◽  
Author(s):  
A. Cyrus Arman ◽  
Alapakkam P. Sampath
Keyword(s):  
Patch Clamp ◽  
Retinal Neurons ◽  
Slice Preparation

scholarly journals Method to remove photoreceptors from whole mount retina in vitro

2017 ◽  
Vol 118 (5) ◽  
pp. 2763-2769 ◽  
Author(s):  
Steven T. Walston ◽  
Yao-Chuan Chang ◽  
James D. Weiland ◽  
Robert H. Chow
Keyword(s):  
Patch Clamp ◽  
Dna Analysis ◽  
Photoreceptor Cells ◽  
Bipolar Cells ◽  
Inner Nuclear Layer ◽  
Retinal Neurons ◽  
Patch Clamp Recording ◽  
Inner Retina ◽  
Whole Mount

Patch clamp recordings of neurons in the inner nuclear layer of the retina are difficult to conduct in a whole mount retina preparation because surrounding neurons block the path of the patch pipette. Vertical slice preparations or dissociated retinal cells provide access to bipolar cells at the cost of severing the lateral connection between neurons. We have developed a technique to remove photoreceptors from the rodent retina that exposes inner nuclear layer neurons, allowing access for patch clamp recording. Repeated application to and removal of filter paper from the photoreceptor side of an isolated retina effectively and efficiently removes photoreceptor cells and, in degenerate retina, hypertrophied Müller cell end feet. Live-dead assays applied to neurons remaining after photoreceptor removal demonstrated mostly viable cells. Patch clamp recordings from bipolar cells reveal responses similar to those recorded in traditional slice and dissociated cell preparations. An advantage of the photoreceptor peel technique is that it exposes inner retinal neurons in a whole mount retina preparation for investigation of signal processing. A disadvantage is that photoreceptor removal alters input to remaining retinal neurons. The technique may be useful for investigations of extracellular electrical stimulation, photoreceptor DNA analysis, and nonpharmacological removal of light input. NEW & NOTEWORTHY This study reports a method for removing photoreceptors from rodent whole mount retina while preserving the architecture of the inner retina. The method enables easier access to the inner retina for studies of neural processing, such as by patch clamp recording.

328 A patch clamp on D2 and D3 receptors on neurons in striatum using slice preparation

1996 ◽  
Vol 25 ◽  
pp. S57
Author(s):  
Taku Amano ◽  
Kumatoshi Ishihara ◽  
Masashi Sasa
Keyword(s):  
Patch Clamp ◽  
Slice Preparation ◽  
D3 Receptors

A lamprey striatal brain slice preparation for patch-clamp recordings

2007 ◽  
Vol 165 (2) ◽  
pp. 251-256 ◽  
Author(s):  
Jesper Ericsson ◽  
Brita Robertson ◽  
Martin A. Wikström
Keyword(s):  
Patch Clamp ◽  
Brain Slice ◽  
Slice Preparation ◽  
Brain Slice Preparation

Reducing Extracellular Cl− Suppresses Dihydropyridine-Sensitive Ca2+ Currents and Synaptic Transmission in Amphibian Photoreceptors

1997 ◽  
Vol 77 (4) ◽  
pp. 2175-2190 ◽  
Author(s):  
Wallace B. Thoreson ◽  
Ron Nitzan ◽  
Robert F. Miller
Keyword(s):  
Synaptic Transmission ◽  
Physiological Role ◽  
Suppressive Effect ◽  
Horizontal Cells ◽  
Retinal Neurons ◽  
Slice Preparation ◽  
Acetoxymethyl Ester ◽  
Electrophysiological Recordings ◽  
Intracellular Ca ◽  
Underlying Mechanisms

Thoreson, Wallace B., Ron Nitzan, and Robert F. Miller. Reducing extracellular Cl− suppresses dihydropyridine-sensitive Ca2+ currents and synaptic transmission in amphibian photoreceptors. J. Neurophysiol. 77: 2175–2190, 1997. A reduction in extracellular chloride suppresses light-evoked currents of second-order retinal neurons (bipolar and horizontal cells) by reducing release of glutamate from photoreceptors. The underlying mechanisms responsible for this action of reduced extracellular Cl− were studied with a combination of electrophysiological recordings from single neurons in a retinal slice preparation and image analyses of intracellular Ca2+ (Fura-2) and pH [2′,7′-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein, acetoxymethyl ester] in dissociated photoreceptors. The results show that reducing extracellular Cl− suppresses a dihydropyridine (DHP)-sensitive Ca2+ current ( I Ca) in photoreceptors. It is proposed that suppression of I Ca results in suppression of photoreceptor neurotransmission. The suppressive effect of low Cl− on I Ca is not due to antagonism by the substituting anion nor is it mediated by changes in extracellular or intracellular pH. We conclude that normal extracellular levels of Cl− are important for maintenance of the voltage-gated Ca2+ channels that support neurotransmission from photoreceptors. Several ideas are presented about the mechanisms by which Cl− supports photoreceptor neurotransmission and the possibility that modulations of Cl− might play a physiological role in the regulation of Ca2+ channels in photoreceptors and, hence, photoreceptor function.

High-voltage electron microscopy of patch-clamped membranes

1987 ◽  
Vol 45 ◽  
pp. 582-583
Author(s):  
F. Sachs ◽  
M. J. Song
Keyword(s):  
Electron Microscopy ◽  
Cell Membrane ◽  
Patch Clamp ◽  
High Voltage Electron Microscopy ◽  
Small Patch ◽  
Cellular Electrophysiology ◽  
Voltage Electron ◽  
Electron Microscopes ◽  
Patch Technique ◽  
Membrane Patch

Cellular electrophysiology has been revolutionized by the introduction of patch clamp techniques. The patch clamp records current from a small patch of the cell membrane which has been sucked into a glass pipette. The membrane patch, a few micons in diameter, is attached to the glass by a seal which is electrically, diffusionally and mechanically tight. Because of the tight electrical seal, the noise level is low enough to record the activity of single ion channels over a time scale extending from 10μs to days. However, although the patch technique is over ten years old, the patch structure is unknown. The patch is inside a glass pipette where it has been impossible to see with standard electron microscopes. We show here that at 1 Mev the glass pipette is transparent and the membrane within can be seen with a resolution of about 30 A.

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