scholarly journals Progress in automating patch clamp cellular physiology

2018 ◽  
Vol 2 ◽  
pp. 239821281877656 ◽  
Author(s):  
Luca A. Annecchino ◽  
Simon R. Schultz
Keyword(s):  
Patch Clamp ◽  
Cellular Mechanisms ◽  
Biologically Relevant ◽  
Synaptic Physiology ◽  
Complex Information ◽  
Electrophysiological Recordings ◽  
Planar Array ◽  
Patch Clamp Electrophysiology ◽  
Neuronal Computation

Patch clamp electrophysiology has transformed research in the life sciences over the last few decades. Since their inception, automatic patch clamp platforms have evolved considerably, demonstrating the capability to address both voltage- and ligand-gated channels, and showing the potential to play a pivotal role in drug discovery and biomedical research. Unfortunately, the cell suspension assays to which early systems were limited cannot recreate biologically relevant cellular environments, or capture higher order aspects of synaptic physiology and network dynamics. In vivo patch clamp electrophysiology has the potential to yield more biologically complex information and be especially useful in reverse engineering the molecular and cellular mechanisms of single-cell and network neuronal computation, while capturing important aspects of human disease mechanisms and possible therapeutic strategies. Unfortunately, it is a difficult procedure with a steep learning curve, which has restricted dissemination of the technique. Luckily, in vivo patch clamp electrophysiology seems particularly amenable to robotic automation. In this review, we document the development of automated patch clamp technology, from early systems based on multi-well plates through to automated planar-array platforms, and modern robotic platforms capable of performing two-photon targeted whole-cell electrophysiological recordings in vivo.

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 PalaCell2D: A framework for detailed tissue morphogenesis

2021 ◽  
Author(s):  
Raphaël Conradin ◽  
Christophe Coreixas ◽  
Jonas Latt ◽  
Bastien Chopard
Keyword(s):  
Internal Pressure ◽  
Tissue Growth ◽  
Cellular Mechanisms ◽  
Simple Description ◽  
Biologically Relevant ◽  
In Vivo Experiments ◽  
Cell Shapes ◽  
And Migration ◽  
The Impact

AbstractIn silico, cell based approaches for modeling biological morphogenesis are used to test and validate our understanding of the biological and mechanical process that are at work during the growth and the organization of multi-cell tissues. As compared to in vivo experiments, computer based frameworks dedicated to tissue modeling allow us to easily test different hypotheses, and to quantify the impact of various biophysically relevant parameters.Here, we propose a formalism based on a detailed, yet simple, description of cells that accounts for intra-, inter- and extra-cellular mechanisms. More precisely, the cell growth and division is described through the space and time evolution of the membrane vertices. These vertices follow a Newtonian dynamics, meaning that their evolution is controlled by different types of forces: a membrane force (spring and bending), an adherence force (inter-cellular spring), external and internal pressure forces. Different evolution laws can be applied on the internal pressure, depending on the intra-cellular mechanism of interest. In addition to the cells dynamics, our formalism further relies on a lattice Boltzmann method, using the Palabos library, to simulate the diffusion of chemical signals. The latter aims at driving the growth and migration of a tissue by simply changing the state of the cells.All of this leads to an accurate description of the growth and division of cells, with realistic cell shapes and where membranes can have different properties. While this work is mainly of methodological nature, we also propose to validate our framework through simple, yet biologically relevant benchmark tests at both single-cell and full tissue scales. This includes free and chemically controlled cell tissue growth in an unbounded domain. The ability of our framework to simulate cell migration, cell compression and morphogenesis under external constraints is also investigated in a qualitative manner.

scholarly journals Robotic automation of “blind” and two-photon targeted patch-clamp electrophysiology in vivo

2021 ◽  
Author(s):  
Gema Vera Gonzalez ◽  
Phatsimo Kgwarae ◽  
Luca Annecchino ◽  
Simon Schultz
Keyword(s):  
Patch Clamp ◽  
Review Paper ◽  
State Of The Art ◽  
Two Photon ◽  
Current State ◽  
Patch Clamp Electrophysiology ◽  
Robotic Automation

A review paper on the current state of the art in robotic automation of in vivo patch-clamp electrophysiology

scholarly journals Robotic Automation of In Vivo Two-Photon Targeted Whole-Cell Patch-Clamp Electrophysiology

Neuron ◽  
2017 ◽  
Vol 95 (5) ◽  
pp. 1048-1055.e3 ◽  
Author(s):  
Luca A. Annecchino ◽  
Alexander R. Morris ◽  
Caroline S. Copeland ◽  
Oshiorenoya E. Agabi ◽  
Paul Chadderton ◽  
...  
Keyword(s):  
Patch Clamp ◽  
Whole Cell ◽  
Two Photon ◽  
Cell Patch Clamp ◽  
Whole Cell Patch Clamp ◽  
Patch Clamp Electrophysiology ◽  
Robotic Automation

Hepatocellular water and electrolyte secretion

1988 ◽  
Vol 66 (10) ◽  
pp. 1253-1260 ◽  
Author(s):  
Christine E. Bear ◽  
Eldon A. Shaffer
Keyword(s):  
Patch Clamp ◽  
Contractile Activity ◽  
Membrane Surface ◽  
Basolateral Membrane ◽  
Experimental Models ◽  
Electrolyte Secretion ◽  
Patch Clamp Electrophysiology ◽  
Ionic Basis ◽  
Water And Electrolyte Secretion

The study of hepatocellular water and electrolyte secretion has been hampered because of the inaccessibility of the hepatobiliary secretory lumen, the canaliculus. The advent of novel experimental models has allowed the application of electrophysiological techniques to investigate the ionic basis of hepatocellular secretion. The "hepatocyte couplet" isolated from the liver in primary monolayer cultures consists of two hepatocytes which enclose a single canalicular unit. The canaliculus of the couplet appears to behave as it would in vivo, exhibiting both secretory and contractile activity. Intracellular microelectrode recordings from this functional unit have permitted direct electrophysiological assessment of cellular and canalicular potentials and measurement of individual ion conductances across the basolateral membrane surface. Further, the application of patch-clamp electrophysiology to study hepatocellular ion transport pathways has characterized individual channel proteins. Intracellular and (or) patch-clamp electrophysiology has elucidated the ion conductance changes activated by bile salts like taurocholate, neurotransmitters like adrenaline, and hormones such as glucagon. These innovative approaches hold much promise in the future study of the ionic basis of hepatocellular secretion.

scholarly journals Robotic automation of “blind” and two-photon targeted patch-clamp electrophysiology in vivo

2021 ◽  
Author(s):  
Gema Vera Gonzalez ◽  
Phatsimo Kgwarae ◽  
Luca Annecchino ◽  
Simon Schultz
Keyword(s):  
Patch Clamp ◽  
Review Paper ◽  
State Of The Art ◽  
Two Photon ◽  
Current State ◽  
Patch Clamp Electrophysiology ◽  
Robotic Automation

A review paper on the current state of the art in robotic automation of in vivo patch-clamp electrophysiology

scholarly journals Automated whole-cell patch-clamp electrophysiology of neurons in vivo

2012 ◽  
Vol 9 (6) ◽  
pp. 585-587 ◽  
Author(s):  
Suhasa B Kodandaramaiah ◽  
Giovanni Talei Franzesi ◽  
Brian Y Chow ◽  
Edward S Boyden ◽  
Craig R Forest
Keyword(s):  
Patch Clamp ◽  
Whole Cell ◽  
Cell Patch Clamp ◽  
Whole Cell Patch Clamp ◽  
Patch Clamp Electrophysiology

scholarly journals Novel 384-Well Population Patch Clamp Electrophysiology Assays for Ca2+-Activated K+ Channels

2006 ◽  
Vol 12 (1) ◽  
pp. 50-60 ◽  
Author(s):  
Victoria H. John ◽  
Tim J. Dale ◽  
Emma C. Hollands ◽  
Mao Xiang Chen ◽  
Leanne Partington ◽  
...  
Keyword(s):  
Patch Clamp ◽  
Rank Order ◽  
Large Cell ◽  
Activator Concentration ◽  
Response Curves ◽  
Channel Expression ◽  
Planar Array ◽  
Patch Clamp Electrophysiology ◽  
Assay Precision ◽  
Voltage Gated Ion Channels

Planar array electrophysiology techniques were applied to assays for modulators of recombinant hIK and hSK3 Ca2+-activated K+ channels. In CHO-hIK—expressing cells, under asymmetric K+ gradients, small-molecule channel activators evoked time- and voltage-independent currents characteristic of those previously described by classical patch clamp electrophysiology methods. In single-hole (cell) experiments, the large cell-to-cell heterogeneity in channel expression rendered it difficult to generate activator concentration-response curves. However, in population patch clamp mode, in which signals are averaged from up to 64 cells, well-to-well variation was substantially reduced such that concentration-response curves could be easily constructed. The absolute EC50 values and rank order of potency for a range of activators, including 1-EBIO and DC-EBIO, corresponded well with conventional patch clamp data. Activator responses of hIK and hSK3 channels could be fully and specifically blocked by the selective inhibitors TRAM-34 and apamin, with IC50 values of 0.31 μM and 3 nM, respectively. To demonstrate assay precision and robustness, a test set of 704 compounds was screened in a 384-well format of the hIK assay. All plates had Z′ values greater than 0.6, and the statistical cutoff for activity was 8%. Eleven hits (1.6%) were identified from this set, in addition to the randomly spiked wells with known activators. Overall, our findings demonstrate that population patch clamp is a powerful and enabling method for screening Ca2+-activated K+ channels and provides significant advantages over single-cell electrophysiology (IonWorksHT) and other previously published approaches. Moreover, this work demonstrates for the 1st time the utility of population patch clamp for ion channel activator assays and for non—voltage-gated ion channels.

scholarly journals Synaptic mechanisms of top-down control in the non-lemniscal inferior colliculus

eLife ◽  
2022 ◽  
Vol 10 ◽  
Author(s):  
Hannah M Oberle ◽  
Alexander N Ford ◽  
Deepak Dileepkumar ◽  
Jordyn Czarny ◽  
Pierre F Apostolides
Keyword(s):  
Perceptual Learning ◽  
Inferior Colliculus ◽  
Auditory Pathway ◽  
Excitatory Postsynaptic Potential ◽  
Descending Pathways ◽  
Subcortical Structures ◽  
Cellular Mechanisms ◽  
Patch Clamp Electrophysiology

Corticofugal projections to evolutionarily ancient, subcortical structures are ubiquitous across mammalian sensory systems. These ‘descending’ pathways enable the neocortex to control ascending sensory representations in a predictive or feedback manner, but the underlying cellular mechanisms are poorly understood. Here, we combine optogenetic approaches with in vivo and in vitro patch-clamp electrophysiology to study the projection from mouse auditory cortex to the inferior colliculus (IC), a major descending auditory pathway that controls IC neuron feature selectivity, plasticity, and auditory perceptual learning. Although individual auditory cortico-collicular synapses were generally weak, IC neurons often integrated inputs from multiple corticofugal axons that generated reliable, tonic depolarizations even during prolonged presynaptic activity. Latency measurements in vivo showed that descending signals reach the IC within 30 ms of sound onset, which in IC neurons corresponded to the peak of synaptic depolarizations evoked by short sounds. Activating ascending and descending pathways at latencies expected in vivo caused a NMDA receptor-dependent, supralinear excitatory postsynaptic potential summation, indicating that descending signals can nonlinearly amplify IC neurons’ moment-to-moment acoustic responses. Our results shed light upon the synaptic bases of descending sensory control and imply that heterosynaptic cooperativity contributes to the auditory cortico-collicular pathway’s role in plasticity and perceptual learning.

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