Recording SOCE Activity in Neurons by Patch-Clamp Electrophysiology and Microfluorometric Calcium Imaging

2018 ◽  
pp. 41-53 ◽  
Author(s):  
Hsiang-en Wu ◽  
Geza Gemes ◽  
Quinn H. Hogan
Keyword(s):  
Patch Clamp ◽  
Calcium Imaging ◽  
Patch Clamp Electrophysiology

scholarly journals Adrenoreceptor modulation of oromotor pathways in the rat medulla

2014 ◽  
Vol 112 (3) ◽  
pp. 580-593 ◽  
Author(s):  
Jason S. Nasse ◽  
Joseph B. Travers
Keyword(s):  
Patch Clamp ◽  
Calcium Imaging ◽  
Solitary Tract ◽  
Noradrenergic Neurons ◽  
Medullary Reticular Formation ◽  
Inward Current ◽  
Postsynaptic Currents ◽  
Presynaptic Mechanisms ◽  
Reticular Neurons ◽  
Patch Clamp Electrophysiology

Regulation of feeding behavior involves the integration of multiple physiological and neurological pathways that control both nutrient-seeking and consummatory behaviors. The consummatory phase of ingestion includes stereotyped oromotor movements of the tongue and jaw that are controlled through brain stem pathways. These pathways encompass not only cranial nerve sensory and motor nuclei for processing feeding-related afferent signals and supplying the oromotor musculature but also reticular neurons for orchestrating ingestion and coordinating it with other behaviors that utilize the same musculature. Based on decerebrate studies, this circuit should be sensitive to satiety mechanisms mediated centrally by A2 noradrenergic neurons in the caudal nucleus of the solitary tract (cNST) that are potently activated during satiety. Because the first observable phase of satiety is inhibition of oromotor movements, we hypothesized that norepinephrine (NE) would act to inhibit prehypoglossal neurons in the medullary reticular formation. Using patch-clamp electrophysiology of retrogradely labeled prehypoglossal neurons and calcium imaging to test this hypothesis, we demonstrate that norepinephrine can influence both pre- and postsynaptic properties of reticular neurons through both α1- and α2-adrenoreceptors. The α1-adrenoreceptor agonist phenylephrine (PE) activated an inward current in the presence of TTX and increased the frequency of both inhibitory and excitatory miniature postsynaptic currents. The α2-adrenoreceptor agonist dexmedetomidine (DMT) inhibited cNST-evoked excitatory currents as well as spontaneous and miniature excitatory currents through presynaptic mechanisms. The diversity of adrenoreceptor modulation of these prehypoglossal neurons may reflect their role in a multifunctional circuit coordinating both ingestive and respiratory lingual function.

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

scholarly journals Functional characterization of transient receptor potential channels in mouse urothelial cells

2010 ◽  
Vol 298 (3) ◽  
pp. F692-F701 ◽  
Author(s):  
Wouter Everaerts ◽  
Joris Vriens ◽  
Grzegorz Owsianik ◽  
Giovanni Appendino ◽  
Thomas Voets ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Patch Clamp ◽  
Calcium Imaging ◽  
Transient Receptor Potential ◽  
Receptor Potential ◽  
Functional Expression ◽  
Sensory Function ◽  
Trp Channel ◽  
Urothelial Cells ◽  
Transient Receptor

The bladder urothelium is currently believed to be a sensory structure, contributing to mechano- and chemosensation in the bladder. Transient receptor potential (TRP) cation channels act as polymodal sensors and may underlie some of the receptive properties of urothelial cells. However, the exact TRP channel expression profile of urothelial cells is unclear. In this study, we have performed a systematic analysis of the molecular and functional expression of various TRP channels in mouse urothelium. Urothelial cells from control and trpv4−/− mice were isolated, cultured (12–48 h), and used for quantitative real-time PCR, immunocytochemistry, calcium imaging, and whole cell patch-clamp experiments. At the mRNA level, TRPV4, TRPV2, and TRPM7 were the most abundantly expressed TRP genes. Immunohistochemistry showed a clear expression of TRPV4 in the plasma membrane, whereas TRPV2 was more prominent in the cytoplasm. TRPM7 was detected in the plasma membrane as well as cytoplasmic vesicles. Calcium imaging and patch-clamp experiments using TRP channel agonists and antagonists provided evidence for the functional expression of TRPV4, TRPV2, and TRPM7 but not of TRPA1, TRPV1, and TRPM8. In conclusion, we have demonstrated functional expression of TRPV4, TRPV2, and TRPM7 in mouse urothelial cells. These channels may contribute to the (mechano)sensory function of the urothelial layer and represent potential targets for the treatment of bladder dysfunction.

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.

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