Relevant bibliographies by topics / Whole-cell patch clamping

Academic literature on the topic 'Whole-cell patch clamping'

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Published: 4 June 2021
Last updated: 3 February 2022

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Journal articles on the topic "Whole-cell patch clamping"

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Garten, Matthias, Lars D. Mosgaard, Thomas Bornschlögl, Stéphane Dieudonné, Patricia Bassereau, and Gilman E. S. Toombes. "Whole-GUV patch-clamping." Proceedings of the National Academy of Sciences 114, no. 2 (December 21, 2016): 328–33. http://dx.doi.org/10.1073/pnas.1609142114.

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Studying how the membrane modulates ion channel and transporter activity is challenging because cells actively regulate membrane properties, whereas existing in vitro systems have limitations, such as residual solvent and unphysiologically high membrane tension. Cell-sized giant unilamellar vesicles (GUVs) would be ideal for in vitro electrophysiology, but efforts to measure the membrane current of intact GUVs have been unsuccessful. In this work, two challenges for obtaining the “whole-GUV” patch-clamp configuration were identified and resolved. First, unless the patch pipette and GUV pressures are precisely matched in the GUV-attached configuration, breaking the patch membrane also ruptures the GUV. Second, GUVs shrink irreversibly because the membrane/glass adhesion creating the high-resistance seal (>1 GΩ) continuously pulls membrane into the pipette. In contrast, for cell-derived giant plasma membrane vesicles (GPMVs), breaking the patch membrane allows the GPMV contents to passivate the pipette surface, thereby dynamically blocking membrane spreading in the whole-GMPV mode. To mimic this dynamic passivation mechanism, beta-casein was encapsulated into GUVs, yielding a stable, high-resistance, whole-GUV configuration for a range of membrane compositions. Specific membrane capacitance measurements confirmed that the membranes were truly solvent-free and that membrane tension could be controlled over a physiological range. Finally, the potential for ion transport studies was tested using the model ion channel, gramicidin, and voltage-clamp fluorometry measurements were performed with a voltage-dependent fluorophore/quencher pair. Whole-GUV patch-clamping allows ion transport and other voltage-dependent processes to be studied while controlling membrane composition, tension, and shape.
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Zhang, W., S. E. Nilson, and S. M. Assmann. "Isolation and Whole-Cell Patch Clamping of Arabidopsis Guard Cell Protoplasts." Cold Spring Harbor Protocols 2008, no. 6 (June 1, 2008): pdb.prot5014. http://dx.doi.org/10.1101/pdb.prot5014.

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Harrison, Reid R., Ilya Kolb, Suhasa B. Kodandaramaiah, Alexander A. Chubykin, Aimei Yang, Mark F. Bear, Edward S. Boyden, and Craig R. Forest. "Microchip amplifier for in vitro, in vivo, and automated whole cell patch-clamp recording." Journal of Neurophysiology 113, no. 4 (February 15, 2015): 1275–82. http://dx.doi.org/10.1152/jn.00629.2014.

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Patch clamping is a gold-standard electrophysiology technique that has the temporal resolution and signal-to-noise ratio capable of reporting single ion channel currents, as well as electrical activity of excitable single cells. Despite its usefulness and decades of development, the amplifiers required for patch clamping are expensive and bulky. This has limited the scalability and throughput of patch clamping for single-ion channel and single-cell analyses. In this work, we have developed a custom patch-clamp amplifier microchip that can be fabricated using standard commercial silicon processes capable of performing both voltage- and current-clamp measurements. A key innovation is the use of nonlinear feedback elements in the voltage-clamp amplifier circuit to convert measured currents into logarithmically encoded voltages, thereby eliminating the need for large high-valued resistors, a factor that has limited previous attempts at integration. Benchtop characterization of the chip shows low levels of current noise [1.1 pA root mean square (rms) over 5 kHz] during voltage-clamp measurements and low levels of voltage noise (8.2 μV rms over 10 kHz) during current-clamp measurements. We demonstrate the ability of the chip to perform both current- and voltage-clamp measurement in vitro in HEK293FT cells and cultured neurons. We also demonstrate its ability to perform in vivo recordings as part of a robotic patch-clamping system. The performance of the patch-clamp amplifier microchip compares favorably with much larger commercial instrumentation, enabling benchtop commoditization, miniaturization, and scalable patch-clamp instrumentation.
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Yantorno, R. E., D. A. Carre, M. Coca-Prados, T. Krupin, and M. M. Civan. "Whole cell patch clamping of ciliary epithelial cells during anisosmotic swelling." American Journal of Physiology-Cell Physiology 262, no. 2 (February 1, 1992): C501—C509. http://dx.doi.org/10.1152/ajpcell.1992.262.2.c501.

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Anisosmotic cell swelling triggers a regulatory volume decrease (RVD) in cell lines derived from human nonpigmented ciliary epithelium. Measurements of cell volume have indicated that the RVD reflects activation of K+ and/or Cl- channels. We have begun to characterize the putative channels by whole cell patch clamping. The results obtained by altering the external K+ and Cl- concentrations and by adding 20-50 microM quinidine or 1 mM Ba2+ indicate that K+ conductances contribute substantially and Cl- conductances contribute very little to the total membrane conductance (GT) under baseline isotonic conditions. Reducing the external osmolality by 20-50% reversibly and reproducibly increased GT by an order of magnitude. Data obtained from ion substitutions and the channel blockers quinidine and 5-nitro-2-(3-phenylpropylamino)-benzoate indicate that most of the hypotonicity-induced conductance reflects stationary Cl(-)-channel activity. The contribution of new K(+)-channel activity was small at intracellular free Ca2+ concentrations of 10 or 200 nM. We conclude that the RVD triggered by bath hypotonicity primarily reflects increased Cl(-)-channel activity.
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Zhu, Michael X. "A well-known potassium channel plays a critical role in lysosomes." Journal of Cell Biology 216, no. 6 (May 16, 2017): 1513–15. http://dx.doi.org/10.1083/jcb.201704017.

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Whole-endolysosome patch clamping presents new opportunities to identify and characterize channels pivotal for these acidic organelles. In this issue (Wang et al., 2017. J. Cell Biol. https://doi.org/10.1083/jcb.201612123), the identification of a role for the large conductance calcium-activated potassium channel brings new thinking about regulation of lysosome membrane potential and function.
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Clark, B., and P. Mobbs. "Transmitter-operated channels in rabbit retinal astrocytes studied in situ by whole-cell patch clamping." Journal of Neuroscience 12, no. 2 (February 1, 1992): 664–73. http://dx.doi.org/10.1523/jneurosci.12-02-00664.1992.

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Holst, Gregory L., William Stoy, Bo Yang, Ilya Kolb, Suhasa B. Kodandaramaiah, Lu Li, Ulf Knoblich, et al. "Autonomous patch-clamp robot for functional characterization of neurons in vivo: development and application to mouse visual cortex." Journal of Neurophysiology 121, no. 6 (June 1, 2019): 2341–57. http://dx.doi.org/10.1152/jn.00738.2018.

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Patch clamping is the gold standard measurement technique for cell-type characterization in vivo, but it has low throughput, is difficult to scale, and requires highly skilled operation. We developed an autonomous robot that can acquire multiple consecutive patch-clamp recordings in vivo. In practice, 40 pipettes loaded into a carousel are sequentially filled and inserted into the brain, localized to a cell, used for patch clamping, and disposed. Automated visual stimulation and electrophysiology software enables functional cell-type classification of whole cell-patched cells, as we show for 37 cells in the anesthetized mouse in visual cortex (V1) layer 5. We achieved 9% yield, with 5.3 min per attempt over hundreds of trials. The highly variable and low-yield nature of in vivo patch-clamp recordings will benefit from such a standardized, automated, quantitative approach, allowing development of optimal algorithms and enabling scaling required for large-scale studies and integration with complementary techniques. NEW & NOTEWORTHY In vivo patch-clamp is the gold standard for intracellular recordings, but it is a very manual and highly skilled technique. The robot in this work demonstrates the most automated in vivo patch-clamp experiment to date, by enabling production of multiple, serial intracellular recordings without human intervention. The robot automates pipette filling, wire threading, pipette positioning, neuron hunting, break-in, delivering sensory stimulus, and recording quality control, enabling in vivo cell-type characterization.
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Zhang, Yanli, Thai Phung, James Dunlop, and Julie Dalziel. "hERG ion channel pharmacology: cell membrane liposomes in porous-supported lipid bilayers compared with whole-cell patch-clamping." European Biophysics Journal 41, no. 11 (August 31, 2012): 949–58. http://dx.doi.org/10.1007/s00249-012-0852-2.

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Kang, Jiesheng, Yongyi Luo, Michelle Searles, and David Rampe. "Observations on conducting whole-cell patch clamping of the hERG cardiac K+channel in pure human serum." Journal of Applied Toxicology 37, no. 4 (August 24, 2016): 445–53. http://dx.doi.org/10.1002/jat.3377.

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Mansell, S. A., C. L. R. Barratt, and S. M. Wilson. "Potassium channel in human sperm identified by whole cell patch clamping plays a role in normal sperm physiology." Fertility and Sterility 98, no. 3 (September 2012): S13. http://dx.doi.org/10.1016/j.fertnstert.2012.07.047.

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Dissertations / Theses on the topic "Whole-cell patch clamping"

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Kodandaramaiah, Suhasa Bangalore. "Robotics for in vivo whole cell patch clamping." Diss., Georgia Institute of Technology, 2012. http://hdl.handle.net/1853/51932.

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Whole-cell patch clamp electrophysiology of neurons in vivo enables the recording of electrical events in cells with great precision, and supports a wide diversity of morphological and molecular analysis experiments important for the understanding of single-cell and network functions in the intact brain. However, high levels of skill are required in order to perform in vivo patching, and the process is time-consuming and painstaking. Robotic systems for in vivo patching would not only empower a great number of neuroscientists to perform such experiments, but would also open up fundamentally new kinds of experiment enabled by the resultant high throughput and scalability. We discovered that in vivo blind whole cell patch clamp electrophysiology could be implemented as a straightforward algorithm and developed an automated robotic system that was capable of performing this algorithm. We validated the performance of the robot in both the cortex and hippocampus of anesthetized mice. The robot achieves yields, cell recording qualities, and operational speeds that are comparable to, or exceed, those of experienced human investigators. Building upon this framework, we developed a multichannel version of “autopatcher” robot capable establishing whole cell patch clamp recordings from pairs and triplets of neurons in the cortex simultaneously. These algorithms can be generalized to control arbitrarily large number of electrodes and the high yield, throughput and automation of complex set of tasks results in a practical solution for conducting patch clamp recordings in potentially dozens of interconnected neurons in vivo.
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Maimaiti, Shaniya. "INSULIN ACTIONS ON HIPPOCAMPAL NEURONS." UKnowledge, 2017. http://uknowledge.uky.edu/pharmacol_etds/20.

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Aging is the main risk factor for cognitive decline. The hippocampus, a brain region critical for learning and memory formation, is especially vulnerable to normal and pathological age-related cognitive decline. Dysregulation of both insulin and intracellular Ca2+ signaling appear to coexist and their compromised actions may synergistically contribute to neuronal dysfunction with aging. This dissertation focused on the interaction between insulin, Ca2+ dysregulation, and cognition in hippocampal neurons by examining the contributions of insulin to Ca2+ signaling events that influence memory formation. I tested the hypothesis that insulin would increase cognition in aged animals by altering Ca2+-dependent physiological mechanisms involved in learning. The possible effects of insulin on learning and memory in young and aged rats were studied. In addition, the effects of insulin on the Ca2+-dependent afterhyperpolarization in CA1 pyramidal hippocampal neurons from young and aged animals were compared. Further, primary hippocampal cultures were used to examine the possible effects of insulin on voltage-gated Ca2+ channel activity and Ca2+-induced Ca2+-release; mechanisms known to influence the AHP. We found that intranasal insulin improved memory in aged F344 rats. Young and aged F344 rats were treated with Humalog®, a short-acting insulin analog, or Levemir®, a long-acting insulin analog. The aged rats performed similar to young rats in the Morris Water Maze, a hippocampal dependent spatial learning and memory task. Electrophysiological recordings from CA1 hippocampal neurons revealed that insulin reduced the age-related increase in the Ca2+-dependent afterhyperpolarization, a prominent biomarker of brain aging that is associated with cognitive decline. Patch clamping recording from hippocampal cultured neurons showed that insulin reduced Ca2+ channel currents. Intracellular Ca2+ levels were also monitored using Fura-2 in response to cellular depolarization. Results indicated that a reduction in Ca2+-induced Ca2+-release from intracellular stores occurred in the presence of insulin. These results suggest that increasing brain insulin levels in aged rats may have improved memory by reducing the AHP and intracellular Ca2+concentrations. This study indicates a possible mechanism responsible for the beneficial effects of intranasal insulin on cognitive function absorbed in selective Alzheimer’s patients. Thus, insulin therapy may reduce or prevent age-related compromises to Ca2+ regulatory pathways typically associated with cognitive decline.
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Dukoff, David. "The Impact of ROS Scavenging on NMDA and AMPA Receptor Whole Cell Currents in Pyramidal Neurons of the Anoxia Tolerant Western Painted Turtle." Thesis, 2013. http://hdl.handle.net/1807/42826.

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Extended periods of oxygen deprivation cause brain death in mammals but the western painted turtle overwinters in anoxic mud for months without damage. Neural protection is achieved through decreases in the whole cell currents of N-methyl-D-aspartate and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (NMDAR and AMPAR) that are dependent on a mild increase in intracellular calcium from the mitochondria. The goal of this research was to determine if natural anoxic decreases in reactive oxidative species (ROS) serve as the signal to bring about these changes. Reductions in cellular ROS levels were demonstrated to have no effect on AMPAR currents or intracellular calcium and produced massive increases in NMDAR currents, indicating that ROS depression does not directly mediate anoxic alterations. Interestingly, mammalian neural tissue also experiences a similar increase in NMDAR whole cell current in response to reducing agents suggesting a possible conserved mechanism for normoxic receptor control.
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Wrigley, Paul John. "Cold thermal processing in the spinal cord." 2006. http://hdl.handle.net/2123/1619.

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Doctor of Philosophy(PhD)
Two recently identified transient receptor potential (TRP) channels, TRPM8 and TRPA1, have been proposed to play an important role in mammalian cool and cold peripheral sensory transduction. When expressed in cell-lines the cloned TRPM8 and TRPA1 receptors have distinct pharmacological and temperature response characteristics. Although these receptors are also transported to the central terminals of primary afferents, little is known about their centrally mediated actions. In this thesis, I use an in vitro electrophysiological approach to investigate the dorsal horn processing of cool afferent modalities and the role of TRP ion channels. The results of this thesis provide further information on thermal processing, indicate direction for further research and suggest possible therapeutic targets for the management of abnormal cold sensory processing. Initial experiments demonstrate that the cooling agents and known TRPM8 and TRPA1 agonists, menthol and icilin, inhibit primary afferent evoked excitatory postsynaptic currents (EPSCs) in rat spinal cord dorsal horn neurons. In addition, temperature reduction, menthol and icilin increase the frequency of miniature EPSCs without affecting amplitude distribution or kinetics. Little or no direct postsynaptic effect on dorsal horn neurons, GABAergic or glycinergic transmission was found. In combination, these observations demonstrate that temperature reduction, menthol and icilin act presynaptically to increase the probability of glutamate release from primary afferent fibres. Further examination of the changes in glutamatergic synaptic transmission induced by temperature reduction, menthol and icilin reveals a subset of neurons sensitive to innocuous cool (< 29 oC) and low concentrations of icilin (3-10 µM) which closely match the temperature activation and pharmacological profile of TRPM8. In addition, the majority of lamina I and II neurons displayed characteristics partly consistent with TRPA1-activation, including a concentration-dependent response to icilin and blockade by ruthenium red. The present experiments did not allow thermal characterisation of these TRPA1-like responses. Together these observations indicate that the effects of menthol and icilin on glutamatergic synaptic transmission in the superficial dorsal horn are mediated by TRPM8 and possibly by TRPA1. Examination of the anatomical location of neurons activated by temperature reduction, menthol, icilin and capsaicin allowed the central termination pattern of thermoreceptive primary afferent fibres with specific TRP-like response characteristics to be determined. TRPM8-like presynaptic activation was confined to a subpopulation of neurons located in lamina I and outer lamina II, while the majority of neurons throughout laminae I and II received inputs sensitive to menthol, high concentrations of icilin and capsaicin. These findings suggest that innocuous cool sensation projects to a specific subpopulation of superficial dorsal horn neurons unlike other modalities (mediated by TRPV1, possibly TRPA1 and other receptors), which non-selectively engage circuits within the entire superficial dorsal horn. No morphological specificity was identified for recovered neurons after electrophysiological characterisation. Finally, mu-opioids were shown to inhibit basal glutamatergic synaptic transmission as well as menthol- and icilin-induced transmission in the superficial dorsal horn. Of particular interest, delta-opioids selectively inhibited icilin-induced synaptic transmission within the same location. The selective effect of delta-opioids suggests a possible role in modulating receptors activated by icilin (TRPM8 and TRPA1). Overall, this thesis provides further evidence that TRPM8 is responsible for the transduction of innocuous cold sensation in mammals and is a potential therapeutic target in humans with cold hyperaesthesia secondary to abnormal thermal processing. The use of delta-opioid agonists warrants further investigation in cold hypersensitivity states and potentially other forms of pain.
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Book chapters on the topic "Whole-cell patch clamping"

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Chen, Xiao-Liang, Jiesheng Kang, and David Rampe. "Manual Whole-Cell Patch-Clamping of the HERG Cardiac K+ Channel." In Methods in Molecular Biology, 151–63. Totowa, NJ: Humana Press, 2010. http://dx.doi.org/10.1007/978-1-60761-849-2_9.

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Van Duijn, Bert, Can Ince, Zheng Wang, A. Freek Weidema, Kees R. Libbenga, and Dirk L. Ypey. "Whole-Cell Patch-Clamping: Introducing Substances into Cells During Electrical Measurements from the Cell Membrane -A Review of Potential Difficulties in Plant and Animal Cells." In Biotechnology Applications of Microinjection, Microscopic Imaging, and Fluorescence, 99–109. Boston, MA: Springer US, 1993. http://dx.doi.org/10.1007/978-1-4615-2828-9_12.

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Lockery, Shawn R., and M. B. Goodman. "[13] Tight-seal whole-cell patch clamping of caenorhabditis elegans neurons." In Methods in Enzymology, 201–17. Elsevier, 1998. http://dx.doi.org/10.1016/s0076-6879(98)93016-6.

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Armstrong, Clay M., and William F. Gilly. "[5] Access resistance and space clamp problems associated with whole-cell patch clamping." In Methods in Enzymology, 100–122. Elsevier, 1992. http://dx.doi.org/10.1016/0076-6879(92)07007-b.

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Conference papers on the topic "Whole-cell patch clamping"

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Zhao, Qili, Yu Han, Yiqing Jia, Ningbo Yu, Mingzhu Sun, and Xin Zhao. "Robotic Whole-cell Patch Clamping Based on Three Dimensional Location for Adherent Cells." In 2020 International Conference on Manipulation, Automation and Robotics at Small Scales (MARSS). IEEE, 2020. http://dx.doi.org/10.1109/marss49294.2020.9307890.

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