High-Throughput Profiling of Ion Channel Activity in Primary Human Lymphocytes

2008 ◽  
Vol 80 (10) ◽  
pp. 3728-3735 ◽  
Author(s):  
Daniel J. Estes ◽  
Sohiel Memarsadeghi ◽  
Steven K. Lundy ◽  
Francesc Marti ◽  
Daniel D. Mikol ◽  
...  
Keyword(s):  
Ion Channel ◽  
High Throughput ◽  
Human Lymphocytes ◽  
Channel Activity

scholarly journals Fluorescence Imaging of Electrically Stimulated Cells

2003 ◽  
Vol 8 (6) ◽  
pp. 660-667 ◽  
Author(s):  
Paul Burnett ◽  
Janet K. Robertson ◽  
Jeffrey M. Palmer ◽  
Richard R. Ryan ◽  
Adrienne E. Dubin ◽  
...  
Keyword(s):  
Membrane Potential ◽  
Ion Channels ◽  
Ion Channel ◽  
High Throughput ◽  
Pharmacological Intervention ◽  
Voltage Gradient ◽  
Channel Activity ◽  
Voltage Gated ◽  
Voltage Gated Ion Channels ◽  
Gated Ion Channels

Designing high-throughput screens for voltage-gated ion channels has been a tremendous challenge for the pharmaceutical industry because channel activity is dependent on the transmembrane voltage gradient, a stimulus unlike ligand binding to G-protein-coupled receptors or ligand-gated ion channels. To achieve an acceptable throughput, assays to screen for voltage-gated ion channel modulators that are employed today rely on pharmacological intervention to activate these channels. These interventions can introduce artifacts. Ideally, a high-throughput screen should not compromise physiological relevance. Hence, a more appropriate method would activate voltage-gated ion channels by altering plasma membrane potential directly, via electrical stimulation, while simultaneously recordingthe operation of the channel in populations of cells. The authors present preliminary results obtained from a device that is designed to supply precise and reproducible electrical stimuli to populations of cells. Changes in voltage-gated ion channel activity were monitored using a digital fluorescent microscope. The prototype electric field stimulation (EFS) device provided real-time analysis of cellular responsiveness to physiological and pharmacological stimuli. Voltage stimuli applied to SK-N-SH neuroblastoma cells cultured on the EFS device evoked membrane potential changes that were dependent on activation of voltage-gated sodium channels. Data obtained using digital fluorescence microscopy suggests suitability of this system for HTS.

Myrsinane and related diterpenes from Euphorbia falcata with selective potassium ion channel activity

Planta Medica ◽  
2015 ◽  
Vol 81 (16) ◽  
Author(s):  
A Vasas ◽  
P Orvos ◽  
L Tálosi ◽  
P Forgo ◽  
G Pinke ◽  
...  
Keyword(s):  
Ion Channel ◽  
Potassium Ion ◽  
Channel Activity ◽  
Potassium Ion Channel

Upscaling and Automation of Electrophysiology: Toward High Throughput Screening in Ion Channel Drug Discovery

2003 ◽  
Vol 9 (1) ◽  
pp. 49-58
Author(s):  
Margit Asmild ◽  
Nicholas Oswald ◽  
Karen M. Krzywkowski ◽  
Søren Friis ◽  
Rasmus B. Jacobsen ◽  
...  
Keyword(s):  
Drug Discovery ◽  
Ion Channel ◽  
High Throughput ◽  
High Throughput Screening

scholarly journals Neuromuscular junction-specific genes screening by deep RNA-seq analysis

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Tiankun Hui ◽  
Hongyang Jing ◽  
Xinsheng Lai
Keyword(s):  
Ion Channel ◽  
Neuromuscular Junctions ◽  
Gene Set Enrichment Analysis ◽  
Differentially Expressed ◽  
Muscle Membrane ◽  
Receptor Interaction ◽  
Specific Gene ◽  
Rna Seq ◽  
Channel Activity ◽  
Ppi Networks

Abstract Background Neuromuscular junctions (NMJs) are chemical synapses formed between motor neurons and skeletal muscle fibers and are essential for controlling muscle contraction. NMJ dysfunction causes motor disorders, muscle wasting, and even breathing difficulties. Increasing evidence suggests that many NMJ disorders are closely related to alterations in specific gene products that are highly concentrated in the synaptic region of the muscle. However, many of these proteins are still undiscovered. Thus, screening for NMJ-specific proteins is essential for studying NMJ and the pathogenesis of NMJ diseases. Results In this study, synaptic regions (SRs) and nonsynaptic regions (NSRs) of diaphragm samples from newborn (P0) and adult (3-month-old) mice were used for RNA-seq. A total of 92 and 182 genes were identified as differentially expressed between the SR and NSR in newborn and adult mice, respectively. Meanwhile, a total of 1563 genes were identified as differentially expressed between the newborn SR and adult SR. Gene Ontology (GO) enrichment analyses, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis and gene set enrichment analysis (GSEA) of the DEGs were performed. Protein–protein interaction (PPI) networks were constructed using STRING and Cytoscape. Further analysis identified some novel proteins and pathways that may be important for NMJ development, maintenance and maturation. Specifically, Sv2b, Ptgir, Gabrb3, P2rx3, Dlgap1 and Rims1 may play roles in NMJ development. Hcn1 may localize to the muscle membrane to regulate NMJ maintenance. Trim63, Fbxo32 and several Asb family proteins may regulate muscle developmental-related processes. Conclusion Here, we present a complete dataset describing the spatiotemporal transcriptome changes in synaptic genes and important synaptic pathways. The neuronal projection-related pathway, ion channel activity and neuroactive ligand-receptor interaction pathway are important for NMJ development. The myelination and voltage-gated ion channel activity pathway may be important for NMJ maintenance. These data will facilitate the understanding of the molecular mechanisms underlying the development and maintenance of NMJ and the pathogenesis of NMJ disorders.

ChemInform Abstract: Oligo-THF Peptides: Synthesis, Membrane Insertion, and Studies of Ion Channel Activity.

ChemInform ◽  
2010 ◽  
Vol 28 (9) ◽  
pp. no-no
Author(s):  
H. WAGNER ◽  
K. HARMS ◽  
U. KOERT ◽  
S. MEDER ◽  
G. BOHEIM
Keyword(s):  
Ion Channel ◽  
Membrane Insertion ◽  
Channel Activity ◽  
Peptides Synthesis

scholarly journals Fluorescent toxins as ion channel activity sensors

2021 ◽  
Author(s):  
Robert Stewart ◽  
Bruce E. Cohen ◽  
Jon T. Sack
Keyword(s):  
Ion Channel ◽  
Channel Activity ◽  
Activity Sensors

scholarly journals Severe Acute Respiratory Syndrome Coronavirus Envelope Protein Ion Channel Activity Promotes Virus Fitness and Pathogenesis

2014 ◽  
Vol 10 (5) ◽  
pp. e1004077 ◽  
Author(s):  
Jose L. Nieto-Torres ◽  
Marta L. DeDiego ◽  
Carmina Verdiá-Báguena ◽  
Jose M. Jimenez-Guardeño ◽  
Jose A. Regla-Nava ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Ion Channel ◽  
Envelope Protein ◽  
Channel Activity ◽  
Virus Fitness

scholarly journals pH-Insensitive FRET Voltage Dyes

2007 ◽  
Vol 12 (5) ◽  
pp. 656-667 ◽  
Author(s):  
Michael P. Maher ◽  
Nyan-Tsz Wu ◽  
Hong Ao
Keyword(s):  
Ion Channel ◽  
High Throughput ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
High Sensitivity ◽  
Fast Response ◽  
Resting Membrane Potential ◽  
Absolute Calibration ◽  
High Throughput Assay ◽  
Voltage Sensitive Dyes

Many high-throughput ion channel assays require the use of voltage-sensitive dyes to detect channel activity in the presence of test compounds. Dye systems employing Förster resonance energy transfer (FRET) between 2 membrane-bound dyes are advantageous in combining high sensitivity, relatively fast response, and ratiometric output. The most widely used FRET voltage dye system employs a coumarin fluorescence donor whose excitation spectrum is pH dependent. The authors have validated a new class of voltage-sensitive FRET donors based on a pyrene moiety. These dyes are significantly brighter than CC2-DMPE and are not pH sensitive in the physiological range. With the new dye system, the authors demonstrate a new high-throughput assay for the acid-sensing ion channel (ASIC) family. They also introduce a novel method for absolute calibration of voltage-sensitive dyes, simultaneously determining the resting membrane potential of a cell. ( Journal of Biomolecular Screening 2007:656-667)

A cell-based impedance assay for monitoring transient receptor potential (TRP) ion channel activity

2011 ◽  
Vol 26 (5) ◽  
pp. 2376-2382 ◽  
Author(s):  
Oliver Pänke ◽  
Winnie Weigel ◽  
Sabine Schmidt ◽  
Anja Steude ◽  
Andrea A. Robitzki
Keyword(s):  
Ion Channel ◽  
Transient Receptor Potential ◽  
Receptor Potential ◽  
Channel Activity ◽  
A Cell ◽  
Transient Receptor ◽  
Trp Ion Channel
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