Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches

1981 ◽  
Vol 391 (2) ◽  
pp. 85-100 ◽  
Author(s):  
O. P. Hamill ◽  
A. Marty ◽  
E. Neher ◽  
B. Sakmann ◽  
F. J. Sigworth
Keyword(s):  
High Resolution ◽  
Patch Clamp ◽  
Free Membrane

scholarly journals Patch-Clamp Proteomics of Single Neuronal Somas in Tissue Using Electrophysiology and Subcellular Capillary Electrophoresis Mass Spectrometry

2021 ◽  
Author(s):  
Sam B. Choi ◽  
Abigail M. Polter ◽  
Peter Nemes
Keyword(s):  
Mass Spectrometry ◽  
Capillary Electrophoresis ◽  
High Resolution ◽  
Patch Clamp ◽  
Dopaminergic Neurons ◽  
Detection Sensitivity ◽  
Substantia Nigra Pars Compacta ◽  
High Resolution Mass ◽  
Patch Clamp Electrophysiology ◽  
Resolution Mass

ABSTRACTUnderstanding of the relationship between cellular function and molecular composition holds a key to next-generational therapeutics but requires measurement of all types of molecules in cells. Developments in sequencing enabled semi-routine measurement of single-cell genomes and transcriptomes, but analytical tools are scarce for detecting diverse proteins in tissue-embedded cells. To bridge this gap for neuroscience research, we report the integration of patch-clamp electrophysiology with subcellular shot-gun proteomics by high-resolution mass spectrometry (HRMS). Recording of electrical activity permitted identification of dopaminergic neurons in the substantia nigra pars compacta. Ca. 20–50% of the neuronal soma content, containing an estimated 100 pg of total protein, was aspirated into the patch pipette filled with ammonium bicarbonate. About ~1 pg of somal protein, or ~0.25% of the total cellular proteome, was analyzed on a custom-built capillary electrophoresis (CE) high-resolution mass spectrometer (HRMS). A series of experiments were conducted to systematically enhance detection sensitivity through refinements in sample processing and detection, allowing us to quantify ~275 different proteins from somal aspirate-equivalent protein digests. From single neurons, patch-clamp proteomics of the soma quantified 91, 80, and 95 different proteins from 3 different dopaminergic neurons, or 157 proteins in total. Quantification revealed detectable proteomic differences between the somal protein samples. Analysis of canonical knowledge predicted rich interaction networks between the proteins. The integration of patch-clamp electrophysiology with subcellular CE-HRMS proteomics expands the analytical toolbox of neuroscience.

Hopping Probe Ion Conductance Microscopy and its Combined Patch-Clamping：A Powerful Tool for Structural and Functional Studies in Neural Nanobiology

2011 ◽  
Vol 694 ◽  
pp. 54-58
Author(s):  
Xin Liang Zhao ◽  
Xiao Liu ◽  
Hu Jie Lu ◽  
Li Ying Ma ◽  
Rui Ling Gao ◽  
...  
Keyword(s):  
High Resolution ◽  
Patch Clamp ◽  
Ion Channel ◽  
Membrane Structures ◽  
Patch Clamp Technique ◽  
Ion Conductance ◽  
Resolution Imaging ◽  
Living Neurons ◽  
Topographic Imaging ◽  
The Relationship

Continuous high-resolution observations of cell membrane would greatly aid the elucidation of the relationship between structure and function and facilitate the study of physiological processing in cell biology. However, high-resolution studying living neuron membrane structures and its functions is still a challenge in current nanobiology. The new developed Hoping Probe Ion Conductance Microscopy (HPICM) is designed for non-contact continuous high-resolution topographic imaging of living cells under physiological conditions. In this review, we concisely introduced the basic operation principle of HPICM and its applications in high spatial resolution imaging of two living neuron cell models, N-type SK-N-SH cells and NGF-differentiated sympathetic neuron-like PC12 cells. Combining HPICM with patch-clamp technique, we further investigated the functional ion-channel of under-differentiated neuron-like PC12 cells and demonstrated that NGF treatment promoted the outgrowth of neurites and increased the activity of TTX-sensitive sodium channel. All these results demonstrate that HPICM combined with patch-clamp technique offers high-resolution topographic imaging of living neurons with non-contact — making HPICM an ideal high-resolution imaging technique not to interact/interfere with living neurons during image acquisition, and provides detailed information about the relationship between membrane structures and ion-channel functions of living neurons at the same time, which has the potential to become a powerful microscopy for in-depth researching in neural nanobiology.

High-Resolution Patch-Clamp Techniques

1985 ◽  
pp. 121-149 ◽  
Author(s):  
Anthony Auerbach ◽  
Frederick Sachs
Keyword(s):  
High Resolution ◽  
Patch Clamp

Recording of single γ-aminobutyrate- and acetylcholine-activated receptor channels translated by exogenous mRNA in Xenopus oocytes

1983 ◽  
Vol 218 (1213) ◽  
pp. 481-484 ◽  
Keyword(s):  
High Resolution ◽  
Patch Clamp ◽  
Xenopus Oocytes ◽  
Optic Lobe ◽  
Single Channel ◽  
Patch Clamp Recording ◽  
Chick Optic Lobe ◽  
Single Channel Currents ◽  
Channel Currents

High resolution (‘giga-seal’) patch clamp recording in Xenopus oocytes was used to measure single channel currents from ACh- and GABA-activated receptors. The proteins that make up these receptors had been translated from mRNA derived from, respectively, denervated cat muscle and chick optic lobe.

High‐resolution scanning patch‐clamp: new insights into cell function

2002 ◽  
Vol 16 (7) ◽  
pp. 748-750 ◽  
Author(s):  
Yuchun Gu ◽  
Julia Gorelik ◽  
Hilmar A. Spohr ◽  
Andrew Shevchuk ◽  
Max J. Lab ◽  
...  
Keyword(s):  
High Resolution ◽  
Patch Clamp ◽  
Cell Function

scholarly journals High-Resolution Scanning Patch Clamp

2013 ◽  
Vol 112 (8) ◽  
pp. 1088-1090 ◽  
Author(s):  
Gail A. Robertson
Keyword(s):  
High Resolution ◽  
Patch Clamp

Biogenic Amine Neurotransmitter Transporters: Just When You Thought You Knew Them

Physiology ◽  
2005 ◽  
Vol 20 (4) ◽  
pp. 225-231 ◽  
Author(s):  
Randy D. Blakely ◽  
Louis J. DeFelice ◽  
Aurelio Galli
Keyword(s):  
Plasma Membrane ◽  
High Resolution ◽  
Patch Clamp ◽  
Signaling Pathways ◽  
Biogenic Amine ◽  
Membrane Transporters ◽  
High Resolution Imaging ◽  
Regulatory Pathways ◽  
Resolution Imaging ◽  
Amine Neurotransmitters

Plasma membrane transporters have long been known to support the reuptake of biogenic amine neurotransmitters following release in the central and peripheral nervous systems. Using high-resolution imaging, patch-clamp and amperometric approaches, as well as molecular manipulations of transporter-regulatory pathways, surprising new details have been uncovered as to how transporters work and are influenced by signaling pathways and psychostimulants.

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