scholarly journals Giant Liposome Preparation for Imaging and Patch-Clamp Electrophysiology

10.3791/50227 ◽  
2013 ◽  
Author(s):  
Marcus D. Collins ◽  
Sharona E. Gordon
Keyword(s):  
Patch Clamp ◽  
Liposome Preparation ◽  
Patch Clamp Electrophysiology ◽  
Giant Liposome

scholarly journals Synthesis and Evaluation of Novel α-Aminoamides Containing Benzoheterocyclic Moiety for the Treatment of Pain

Molecules ◽  
2021 ◽  
Vol 26 (6) ◽  
pp. 1716
Author(s):  
Kun Tong ◽  
Ruotian Zhang ◽  
Fengzhi Ren ◽  
Tao Zhang ◽  
Junlin He ◽  
...  
Keyword(s):  
Ion Channels ◽  
Patch Clamp ◽  
Formalin Test ◽  
Sodium Ion ◽  
Inhibitory Potency ◽  
Voltage Gated ◽  
Patch Clamp Electrophysiology ◽  
Sodium Ion Channels

Novel α-aminoamide derivatives containing different benzoheterocyclics moiety were synthesized and evaluated as voltage-gated sodium ion channels blocks the treatment of pain. Compounds 6a, 6e, and 6f containing the benzofuran group displayed more potent in vivo analgesic activity than ralfinamide in both the formalin test and the writhing assay. Interestingly, they also exhibited potent in vitro anti-Nav1.7 and anti-Nav1.8 activity in the patch-clamp electrophysiology assay. Therefore, compounds 6a, 6e, and 6f, which have inhibitory potency for two pain-related Nav targets, could serve as new leads for the development of analgesic medicines.

scholarly journals Isolating Nuclei from Cultured Cells for Patch-Clamp Electrophysiology of Intracellular Ca 2+ Channels

2013 ◽  
Vol 2013 (9) ◽  
pp. pdb.prot073056 ◽  
Author(s):  
Don-On Daniel Mak ◽  
Horia Vais ◽  
King-Ho Cheung ◽  
J. Kevin Foskett
Keyword(s):  
Patch Clamp ◽  
Cultured Cells ◽  
Intracellular Ca ◽  
Patch Clamp Electrophysiology

LabPatch, an acquisition and analysis program for patch-clamp electrophysiology

2000 ◽  
Vol 278 (5) ◽  
pp. C1055-C1061 ◽  
Author(s):  
Tim Robinson ◽  
Lars Thomsen ◽  
Jan D. Huizinga
Keyword(s):  
Patch Clamp ◽  
Data Acquisition ◽  
Front Panel ◽  
Analysis Program ◽  
Digital To Analog Converter ◽  
Analog Converter ◽  
Data Acquisition Card ◽  
System Requirements ◽  
User Need ◽  
Patch Clamp Electrophysiology

An acquisition and analysis program, “LabPatch,” has been developed for use in patch-clamp research. LabPatch controls any patch-clamp amplifier, acquires and records data, runs voltage protocols, plots and analyzes data, and connects to spreadsheet and database programs. Controls within LabPatch are grouped by function on one screen, much like an oscilloscope front panel. The software is mouse driven, so that the user need only point and click. Finally, the ability to copy data to other programs running in Windows 95/98, and the ability to keep track of experiments using a database, make LabPatch extremely versatile. The system requirements include Windows 95/98, at least a 100-MHz processor and 16 MB RAM, a data acquisition card, digital-to-analog converter, and a patch-clamp amplifier. LabPatch is available free of charge at http://www.fhs.mcmaster.ca/huizinga/ .

scholarly journals An assessment of KIR channel function in human cerebral arteries

2019 ◽  
Vol 316 (4) ◽  
pp. H794-H800 ◽  
Author(s):  
Maria Sancho ◽  
Yuan Gao ◽  
Bjorn O. Hald ◽  
Hao Yin ◽  
Melfort Boulton ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Patch Clamp ◽  
Cerebral Arteries ◽  
Channel Expression ◽  
Expression Activity ◽  
Patch Clamp Electrophysiology ◽  
Resting Tone ◽  
The Impact ◽  
Arterial Tone

In the rodent cerebral circulation, inward rectifying K+ (KIR) channels set resting tone and the distance over which electrical phenomena spread along the arterial wall. The present study sought to translate these observations into human cerebral arteries obtained from resected brain tissue. Computational modeling and a conduction assay first defined the impact of KIR channels on electrical communication; patch-clamp electrophysiology, quantitative PCR, and immunohistochemistry then characterized KIR2.x channel expression/activity. In keeping with rodent observations, computer modeling highlighted that KIR blockade should constrict cerebral arteries and attenuate electrical communication if functionally expressed. Surprisingly, Ba2+ (a KIR channel inhibitor) had no effect on human cerebral arterial tone or intercellular conduction. In alignment with these observations, immunohistochemistry and patch-clamp electrophysiology revealed minimal KIR channel expression/activity in both smooth muscle and endothelial cells. This absence may be reflective of chronic stress as dysphormic neurons, leukocyte infiltrate, and glial fibrillary acidic protein expression was notable in the epileptic cortex. In closing, KIR2.x channel expression is limited in human cerebral arteries from patients with epilepsy and thus has little impact on resting tone or the spread of vasomotor responses. NEW & NOTEWORTHY KIR2.x channels are expressed in rodent cerebral arterial smooth muscle and endothelial cells. As they are critical to setting membrane potential and the distance signals conduct, we sought to translate this work into humans. Surprisingly, KIR2.x channel activity/expression was limited in human cerebral arteries, a paucity tied to chronic brain stress in the epileptic cortex. Without substantive expression, KIR2.x channels were unable to govern arterial tone or conduction.

scholarly journals Interferon γ Gene Expression in Sensory Neurons: Evidence for Autocrine Gene Regulation

1997 ◽  
Vol 186 (12) ◽  
pp. 2023-2031 ◽  
Author(s):  
Harald Neumann ◽  
Hannes Schmidt ◽  
Elke Wilharm ◽  
Lüder Behrens ◽  
Hartmut Wekerle
Keyword(s):  
Patch Clamp ◽  
Sensory Neurons ◽  
Nuclear Translocation ◽  
Interferon Γ ◽  
Dorsal Root Ganglion Neurons ◽  
Neuronal Membrane ◽  
Cultured Neurons ◽  
Ifn Γ ◽  
Patch Clamp Electrophysiology ◽  
Ganglion Neurons

We explored expression and possible function of interferon-γ (IFN-γ) in cultured fetal (E15) rat dorsal root ganglion neurons combining whole cell patch-clamp electrophysiology with single cell reverse transcriptase polymerase chain reaction and confocal laser immunocytochemistry. Morphologically, we located IFN-γ protein in the cytoplasm of the neurons in culture as well as in situ during peri- and postnatal development. Transcripts for classic IFN-γ and for its receptor were determined in probes of cytoplasm sampled from individual cultured neurons, which had been identified by patch clamp electrophysiology. In addition, the cultured neurons expressed both chains of the IFN-γ receptor. Locally produced IFN-γ acts back on its cellular source. Phosphorylation and nuclear translocation of the IFN-inducible transcriptional factor STAT1 as well as IFN-γ–dependent expression of major histocompatibility complex class I molecules on the neuronal membrane were noted in untreated cultures. However, both processes were substantially blocked in the presence of antibodies neutralizing IFN-γ. Our findings indicate a role of IFN-γ in autocrine regulation of sensory neurons.

scholarly journals 793Validation of cardiac optogenetics against patch clamp electrophysiology in human cardiomyocyte drug efficacy and safety studies

EP Europace ◽  
2017 ◽  
Vol 19 (suppl_3) ◽  
pp. iii141-iii141
Author(s):  
S. Bjork ◽  
E. Ojala ◽  
T. Nordstrom ◽  
E. Kankuri ◽  
E. Mervaala
Keyword(s):  
Patch Clamp ◽  
Drug Efficacy ◽  
Efficacy And Safety ◽  
Safety Studies ◽  
Patch Clamp Electrophysiology

Cultured Insect Mushroom Body Neurons Express Functional Receptors for Acetylcholine, GABA, Glutamate, Octopamine, and Dopamine

1999 ◽  
Vol 81 (1) ◽  
pp. 1-14 ◽  
Author(s):  
M. Cayre ◽  
S. D. Buckingham ◽  
S. Yagodin ◽  
D. B. Sattelle
Keyword(s):  
Patch Clamp ◽  
Mushroom Body ◽  
Gaba Receptor ◽  
Time Course ◽  
Reversal Potential ◽  
Acheta Domesticus ◽  
Free Calcium ◽  
Kenyon Cells ◽  
Inward Currents ◽  
Patch Clamp Electrophysiology

Cayre, M., S. D. Buckingham, S. Yagodin, and D. B. Sattelle. Cultured insect mushroom body neurons express functional receptors for acetylcholine, GABA, glutamate, octopamine, and dopamine. J. Neurophysiol. 81: 1–14, 1999. Fluorescence calcium imaging with fura-2 and whole cell, patch-clamp electrophysiology was applied to cultured Kenyon cells (interneurons) isolated from the mushroom bodies of adult crickets ( Acheta domesticus) to demonstrate the presence of functional neurotransmitter receptors. In all cells investigated, 5 μM acetylcholine (ACh, n = 52) evoked an increase in intracellular free calcium ([Ca2+]i). Similar effects were observed in response to 10 μM nicotine. The ACh response was insensitive to atropine (50 μM) but was reduced by mecamylamine (50 μM) and α-bungarotoxin (α-bgt, 10 μM). ACh-induced inward ion currents ( n = 28, E ACh ∼0 mV) were also blocked by 1 μM mecamylamine and by 1 μM α-bgt. Nicotine-induced inward currents desensitized more rapidly than ACh responses. Thus functional α-bgt–sensitive nicotinic ACh receptors are abundant on all Kenyon cells tested, and their activation leads to an increase in [Ca2+]i. γ-Aminobutyric acid (GABA, 100 μM) triggered a sustained decrease in [Ca2+]i. Similar responses were seen with a GABAA agonist, muscimol (100 μM), and a GABAB agonist, 3-APPA (1 mM), suggesting that more than one type of GABA receptor can affect [Ca2+]i. This action of GABA was not observed when the extracellular KCl concentration was lowered. All cells tested ( n = 26) with patch-clamp electrophysiology showed picrotoxinin (PTX)-sensitive, GABA-induced (30–100 μM) currents with a chloride-sensitive reversal potential. Thus, an ionotropic PTX-sensitive GABA receptor was found on all Kenyon cells tested. Most (61%) of the 54 cells studied responded to l-glutamate (100 μM) application either with a biphasic increase in [Ca2+]i or with a single, delayed, sustained [Ca2+]i increase. Nearly all cells tested (95%, n = 19) responded to (100 μM) l-glutamate with rapidly desensitizing, inward currents that reversed at approximately −30 mV. Dopamine (100 μM) elicited either a rapid or a delayed increase in [Ca2+]i in 63% of the 26 cells tested. The time course of these responses varied greatly among cells. Dopamine failed to elicit currents in patch-clamped cells ( n = 4). A brief decrease in [Ca2+]i was induced by octopamine (100 μM) in ∼54% of the cells tested ( n = 35). However, when extracellular CaCl2 was lowered, octopamine triggered a substantial increase in [Ca2+]i in 35% of the cells tested ( n = 26). No octopamine-elicited currents were detected in patched-clamped cells ( n = 10).

A methodological comparison of human α4β2 and α3β4 receptor properties using conventional and high-throughput patch-clamp electrophysiology techniques

2011 ◽  
Vol 82 (8) ◽  
pp. 1025
Author(s):  
Lisa C. Benson ◽  
Serguei S. Sidach ◽  
John D. Graef ◽  
Patrick M. Lippiello ◽  
Merouane Bencherif ◽  
...  
Keyword(s):  
Patch Clamp ◽  
High Throughput ◽  
Patch Clamp Electrophysiology ◽  
Methodological Comparison
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