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Inhibitory Postsynaptic Currents

Inhibitory Postsynaptic Currents are the electrical signals generated by the activation of inhibitory receptors at the postsynaptic membrane.
These currents serve to reduce the excitability of the postsynaptic neuron, playing a crucial role in regulating neural network activity and information processing in the brain.
Studying Inhibitory Postsynaptic Currents is essential for understanding the balance between excitation and inhibition, which is critical for normal brain function.
Researchers can now leverage the power of AI-driven platforms like PubCompare.ai to optimize their Inhibitory Postsynaptic Currents research, locating the best protocols from literature, preprints, and patents, and enhacing reproducibility and accuracy through intelligent comparisons.
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Most cited protocols related to «Inhibitory Postsynaptic Currents»

GABA-A Receptor Characterization in Neuronal Slices

Cited 13 times

Slices were perfused with ACSF, which was heated to 35–37°C, equilibrated with 95% O₂/5% CO₂ and contained (in mM): 126 NaCl, 26 NaHCO₃, 3 KCl, 1.25
NaH₂PO₄, 1.6 CaCl₂, 1.5 MgSO₄, 10 glucose, 0.05 D-(–)-2-amino-5-phosphonopentanoic acid (APV), 0.02 6,7-dinitroquinoxaline-2,3-dione (DNQX) and 0.002 (2S)-3-{[(1S)-1-(3,4-dichlorophenyl)ethyl]amino-2-hydroxypropyl)(phenylmethyl)phosphinic acid (CGP 55845). APV, DNQX and CGP55845 were used to block NMDA, AMPA/kainate and GABA_B receptors, respectively, so that GABA_A receptor-mediated currents could be studied in relative isolation (Bevan et al., 2002 (link); Hallworth and Bevan, 2005 (link)). Miniature inhibitory postsynaptic currents (mIPSCs) were recorded in the additional presence of 0.5 µM tetrodotoxin. In some cases sulpiride (2 µM) was added to block D2 dopamine receptors. Drugs were purchased from Abcam except for sulpiride, which was obtained from Tocris.
Somatic patch clamp recordings were obtained under visual guidance (Axioskop FS2, Zeiss) using computer-controlled manipulators (Luigs & Neumann) and a Multiclamp 700B amplifier and digidata 1440A digitizer controlled by PClamp 10 (Molecular Devices). Pipettes contained (in mM): 135 CsCl, 3.6 NaCl, 1 MgCl₂, 10 HEPES, 10 QX-314, 0.1 Na₄EGTA, 0.4 Na₃GTP and 2 Mg_1.5ATP (pH 7.2, 290 mOsm) or 130 Kgluconate, 3.6 Nagluconate, 1 MgCl₂, 10 HEPES, 10 QX-314, TEA-Cl 5, 0.1 Na₄EGTA, 0.4 Na₃GTP and 2 Mg_1.5ATP (pH 7.2, 290 mOsm) for the recording of GABA_A receptor-mediated mIPSCs and evoked currents, respectively. mIPSCs were recorded at −60 mV. Evoked IPSCs and isoguvacine-evoked current were recorded at −50 mV. Weighted decay kinetics ofmIPSCs were calculated from τ decay = (A1*τ1 + A2*τ2)/(A1 + A2) where A and τ refer to the amplitude and decay constants, respectively, of biexponential fits of mIPSCs. Data were analyzed with Clampfit 10 (Molecular Devices), Igor Pro 6 (Wavemetrics) and Origin 8 (OriginLab).

Fan K.Y., Baufreton J., Surmeier D.J., Chan C.S, & Bevan M.D. (2012). Proliferation of External Globus Pallidus-Subthalamic Nucleus Synapses following Degeneration of Midbrain Dopamine Neurons. The Journal of Neuroscience, 32(40), 13718-13728.

Publication 2012

6,7-dinitroquinoxaline-2,3-dione alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid Amino Acids Bicarbonate, Sodium Cardiac Arrest cesium chloride CGP 55845 CGP55845 Diploid Cell Dopamine D2 Receptor GABA-A Receptor Glucose HEPES Hypromellose Induced Pluripotent Stem Cells Inhibitory Postsynaptic Currents isoguvacine isolation Kainate Kinetics Magnesium Chloride Medical Devices N-Methylaspartate Pharmaceutical Preparations Phosphinic Acids QX-314 Seizures Sodium Chloride Sulfate, Magnesium Sulpiride Tetrodotoxin

Investigating Neuronal Electrophysiology and Synaptic Dynamics

Cited 7 times

The neurons were recorded by MultiClamp-700B amplifier under voltage-clamp for their synaptic activity and the current-clamp for their intrinsic property. Electrical signals were inputted to pClamp-10 (Axon Instrument Inc.) for data acquisition and analysis. An output bandwidth of the amplifier was set at 3kHz. The pipette solution for recording excitatory events included (mM) 150 K-gluconate, 5 NaCl, 5 HEPES, 0.4 EGTA, 4 Mg-ATP, 0.5 Tris-GTP, and 5 phosphocreatine (pH 7.35; Ge et al., 2011 (link); Yang et al., 2014 (link)). The solution for studying inhibitory synapses contained (mM) 130 K-gluconate, 20 KCl, 5 NaCl, 5 HEPES, 0.5 EGTA, 4 Mg-ATP, 0.5 Tris–GTP, and 5 phosphocreatine (F. Zhang et al., 2012 (link)). These pipette solutions were freshly made and filtered (0.1 μm). The osmolarity was 295 to 305 mOsmol and pipette resistance was 5 to 6 MΩ.
The functions of GABAergic neurons were assessed including their active intrinsic properties and inhibitory outputs (J.-H. Wang, 2003 (link)). The inhibitory outputs were assessed by recording spontaneous inhibitory postsynaptic currents (sIPSC) on glutamatergic neurons in the presence of 10 μM 6-Cyano-7-nitroquinoxaline-2,3-dione and 40 µM D-amino-5-phosphonovanolenic acid in the ACSF to block ionotropic glutamatergic receptors. A total of 10 µM bicuculline was washed onto the slices at the end of experiments for blocking sIPSCs to test that synaptic responses were mediated by GABA_AR. The pipette solution with a high concentration of chloride ions makes the reversal potential -42 mV. sIPSCs are inward when membrane potential is held at -65 mV (Wei et al., 2004 (link); F. Zhang et al., 2012 (link)).
The functions of excitatory neurons were evaluated based on their active intrinsic properties and excitatory output (J.-H. Wang, 2003 (link)). The excitatory outputs were assessed by recording spontaneous excitatory postsynaptic currents (sEPSC) on GABAergic neurons in the presence of 10 µM bicuculline in the ACSF to block GABA_AR (J.-H. Wang, 2003 (link); Yu et al., 2012 (link)). A total of 10 μM 6-cyano-7-nitroquinoxaline-2,3-dione and 40 µM D-amino-5-phosphonovanolenic acid were added into the ACSF at the end of experiments to test whether synaptic responses were mediated by glutamate receptor, which blocked sEPSCs in our studies.
The recording of spontaneous synaptic currents, instead of evoked synaptic currents, is based on the following reasons. sEPSC and sIPSC amplitudes represent the responsiveness and densities of postsynaptic receptors. The frequencies imply the probability of transmitter release from an axon terminal and the number of presynaptic axons innervated on the recorded neuron (Zucker and Regehr, 2002 (link); Stevens, 2004 (link)). Such parameters can be used to analyze presynaptic and postsynaptic mechanisms as well as to compare them with morphological data about neuronal interaction. The evoked postsynaptic currents cannot separate these mechanisms. We did not use tetrodotoxin in the ACSF to record miniature postsynaptic currents, since we had to record neuronal excitability. As the frequency of synaptic activities was less than those of sequential spikes (Figures 2, 4–5) and spontaneous spikes were never recorded on the neurons in our cortical slices, sIPSCs and sEPSCs were not generated from spontaneous action potentials. Synaptic events in our recording are presumably miniature postsynaptic currents. This point is granted by a single peak of postsynaptic currents in our study.
Action potentials at the cortical neurons were induced by injecting the depolarization pulse. Their excitability was assessed by input-outputs (spikes vs normalized stimuli) when various stimuli were given (Chen et al., 2006 (link)). We did not measure rheobase to show cellular excitability, as this strength-duration relationship was used to assess the ability to fire single spike. We measured the ability of firing sequential spikes (J. H. Wang et al., 2008 (link)).
Data were analyzed if the recorded neurons had the resting membrane potentials negatively more than -60 mV and action potential amplitudes more than 90 mV. The criteria for the acceptance of each experiment also included <5% changes in resting membrane potential, spike magnitude, and input resistance throughout each recording. The series and input resistances in all neurons were monitored by injecting hyperpolarization pulses (5 mV/50ms) and calculated by voltage pulses vs instantaneous and steady-state currents.

Xu A., Cui S, & Wang J.H. (2015). Incoordination among Subcellular Compartments Is Associated with Depression-Like Behavior Induced by Chronic Mild Stress. International Journal of Neuropsychopharmacology, 19(5), pyv122.

Publication 2015

Action Potentials Amino Acids ARID1A protein, human Axon Bicuculline Cells Chlorides Cortex, Cerebral Egtazic Acid Electricity Excitatory Postsynaptic Currents GABAergic Neurons gluconate Glutamate Receptor HEPES Inhibitory Postsynaptic Currents Ions Membrane Potentials Neurons Osmolarity Phosphocreatine Post-Synaptic Density Postsynaptic Current Psychological Inhibition Pulse Rate Pulses Resting Potentials Sodium Chloride Synapses Tetrodotoxin Tromethamine

Analysis of CeA GABA Transmission in Alcohol Dependence

Cited 5 times

Preparation of acute brain slices and electrophysiological recordings were performed as previously described^{48 (link)} from a subset of 24-hr abstinent rats from each group that were under deep isoflurane anesthesia. Elevated CeA GABA transmission represents a hallmark of alcohol dependence across species^{10 (link), 11 (link)}, and acute ethanol application increase CeA GABA signaling^{48 (link)}. Thus, we recorded pharmacologically-isolated GABA_A receptor mediated miniature inhibitory postsynaptic currents (mIPSCs) from 51 neurons in the medial subdivision of the CeA using whole-cell voltage clamp mode. Data were analyzed using Mini Analysis (Synaptosoft Inc., Fort Lee, NJ) with 3-min bins of gap-free recording^{48 (link)} and only currents >5 pA accepted for analysis.

Steinman M.Q., Kirson D., Wolfe S.A., Khom S., D’Ambrosio S.R., Bagsic S.R., Bajo M., Vlkolinský R., Hoang N.K., Singhal A., Sureshchandra S., Oleata C.S., Messaoudi I., Zorrilla E.P, & Roberto M. (2020). Importance of sex and trauma context on circulating cytokines and amygdalar GABAergic signaling in a comorbid model of posttraumatic stress and alcohol use disorders. Molecular psychiatry, 26(7), 3093-3107.

Publication 2020

Alcoholic Intoxication, Chronic Anesthesia Brain Cells Ethanol GABA-A Receptor gamma Aminobutyric Acid Inhibitory Postsynaptic Currents Isoflurane Neurons Rattus norvegicus Transmission, Communicable Disease

Electrophysiological Characterization of GABA Signaling in Rat Amygdala

Cited 5 times

Dependent and nondependent rats were deeply anesthetized with isoflurane followed by rapid decapitation and immediate removal of the brain into an ice-cold high-sucrose brain slice cutting solution (sucrose 206 mM; KCl 2.5 mM; CaCl2 0.5 mM; MgCl2 7 mM; NaH2PO4 1.2 mM; NaHCO3 26 mM; glucose 5 mM; HEPES 5 mM [pH 7.4]). Coronal slices (300 to 400 μm) containing the CeA were continuously superfused (flow rate of 2 to 4 ml/min) with 95% O₂/5% CO₂ equilibrated aCSF of the following composition: NaCl 130 mM, KCl 3.5 mM, NaH2PO4 1.25 mM, MgSO4·7H2O 1.5 mM, CaCl2 2.0 mM, NaHCO3 24 mM, and glucose 10 mM. Recordings were performed in neurons from the medial subdivision of the CeA. Each experimental group contained neurons from a minimum of 3 rats. GABAergic activity was pharmacologically isolated with DNQX, DL-AP5, and CGP. All drugs were applied by bath superfusion.
We recorded with sharp micropipettes filled with 3M KCl and evoked GABAergic inhibitory postsynaptic potentials (eIPSPs) by stimulating locally within the medial subdivision of CeA through a bipolar electrode. Neurons were held near their resting membrane potential (−82.4 ± 0.8 mV). We performed an input–output (I/O) protocol consisting of a range of 5 current stimulations, starting at the threshold current required to elicit an eIPSP, up to the strength required to elicit the maximum subthreshold amplitude. The middle stimulus intensity was used to monitor drug-induced changes throughout the duration of the experiment. Paired-pulse ratio (PPR) was performed at the stimulus intensity giving approximately 50% of the maximal amplitude determined in the I/O protocol.
Whole-cell voltage-clamp recordings of GABAergic spontaneous inhibitory postsynaptic currents (sIPSCs) and miniature inhibitory postsynaptic currents (mIPSCs) were from visualized CeA neurons clamped at −60 mV for the duration of the recordings. Patch pipettes (3 to 6 MΩ) were filled with an internal solution composed of the following (in mM): 145 KCl, 0.5 EGTA, 2 MgCl2, 10 HEPES, 2 Na-ATP, and 0.2 Na-GTP. In all experiments, cells with a series resistance greater than 25 MΩ were excluded from analysis, and series resistance was continuously monitored during gap-free recording with a 10-mV pulse. Cells in which series resistance changed more than 25% during the course of the experiment were excluded from analysis. All measures were performed prior to (baseline) and during drug application (details in S1 Text).

Tunstall B.J., Kirson D., Zallar L.J., McConnell S.A., Vendruscolo J.C., Ho C.P., Oleata C.S., Khom S., Manning M., Lee M.R., Leggio L., Koob G.F., Roberto M, & Vendruscolo L.F. (2019). Oxytocin blocks enhanced motivation for alcohol in alcohol dependence and blocks alcohol effects on GABAergic transmission in the central amygdala. PLoS Biology, 17(4), e2006421.

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Publication 2019

6,7-dinitroquinoxaline-2,3-dione ARID1A protein, human Bath Bicarbonate, Sodium Brain Cells Cold Temperature Decapitation Egtazic Acid Glucose HEPES Inhibitory Postsynaptic Currents Inhibitory Postsynaptic Potentials Isoflurane Magnesium Chloride Membrane Potentials Neurons Pharmaceutical Preparations Pulse Rate Rattus Sodium Chloride Sucrose Sulfate, Magnesium

Electrophysiological Recordings and Pharmacological Manipulations

Cited 5 times

Single and dual voltage-clamp recordings were performed using the whole-cell configuration of the patch-clamp technique at a pipette voltage of −60 mV using the Axopatch 200B and 1D amplifiers (Molecular Device Co., Sunnyvale CA, USA). Access resistance was monitored during the recordings, and experiments with >20% change were discarded. The baseline membrane potential for current-clamp recordings was set at −70 mV before each series of current step injection protocols. Rheobase current was defined as the first current step, within a series of increasing 20 pA steps, that elicited an action potential.
Stock solutions of bicuculline methobromide (BMR), tetrodotoxin (TTX), 4,5,6,7-tetrahydroisoxazolo{5,4-c}pyridine-3-ol, (THIP), SKF-81297, quinpirole, sulpiride, SCH 23390, and GABA (all from Sigma) were prepared in water. Etomidate (Sigma) was dissolved in dimethylsulfoxide (<0.0001% final concentration). All stock solutions were diluted to the desired concentration in aCSF and applied locally through a Y tube (Murase et al., 1989 (link)) modified for optimal solution exchange in brain slices (Hevers and Luddens, 2002 ).
Currents were filtered at 2 kHz with a low-pass Bessel filter and digitized at 5–10 kHz using a personal computer equipped with Digidata 1322A data acquisition board and pCLAMP9 software (both from Molecular Devices). Off-line data analysis, curve fitting, and figure preparation were performed with Clampfit 9 software (Molecular Devices). Spontaneous and miniature inhibitory postsynaptic currents (sIPSCs and mIPSCs) were identified using a semi-automated threshold based mini detection software (Mini Analysis, Synaptosoft Inc., Fort Lee, NJ) and were visually confirmed as in Ade et al. (2008) (link). Briefly, IPSC averages were based on >60 events and the decay kinetics were determined using double exponential curve fittings and reported as weighted time constants (tau). All detected events were used for event frequency analysis, but superimposing events were eliminated for the amplitude, rise time, and decay kinetic analysis. Tonic current measurements were made as in Ade et al., 2008 (link). Briefly, an all-points histogram was plotted for a 10 s period immediately before and during BMR application. Tonic currents are represented as the change in baseline amplitude. When PKA or PKI was included in the internal solution, events were analyzed at least four minutes after break-in to allow the peptide to function and equilibrate with the internal components of the cell.
Statistical significance was determined using the two-tailed Student’s t test (unpaired when comparing two populations of cells and paired when comparing results within the same cell). All values are expressed as mean ± SEM. In all figures, *p < 0.05, **p < 0.005, and ***p < 0.0005.

Janssen M.J., Ade K.K., Fu Z, & Vicini S. (2009). Dopamine Modulation of GABA Tonic Conductance in Striatal Output Neurons. The Journal of Neuroscience, 29(16), 5116-5126.

Publication 2009

Action Potentials bicuculline methobromide Brain Cells Cellular Structures Etomidate gaboxadol gamma Aminobutyric Acid Induced Pluripotent Stem Cells Inhibitory Postsynaptic Currents Kinetics Medical Devices Membrane Potentials Peptides Population Group pyridine Quinpirole SCH 23390 SK&F 81297 Student Sulfoxide, Dimethyl Sulpiride Tetrodotoxin

Most recents protocols related to «Inhibitory Postsynaptic Currents»

Hippocampal CA1 Neuron Inhibitory Currents

Single slices were transferred to a recording chamber that was constantly perfused (∼3 mL/min) with oxygenated aCSF at 35 °C. The CA1 neurons were visualized under a Zeiss upright microscope (40 X water-immersion objective) and an enhanced differential interference contrast (DIC) video microscope system. Recording pipettes with the resistance of 3–5 MΩ were pulled from borosilicate glass capillaries (1.5 mm outer diameter) using a P97 electrode puller (Sutter Instruments, Novato, CA). Access resistance and input capacitance were electronically compensated by ∼60–70% and monitored throughout the experiment to confirm the stability of the recording.
Fast spontaneous inhibitory postsynaptic currents (sIPSCs) mediated by the GABA_A receptor were recorded from hippocampal CA1 pyramidal neurons held at −70 mV in the presence of 2-amino-5-phosphonovaleric acid (APV)(50 μM), 6-cyano-7-nitroquinoxaline-2,3-dione (DNQX) (20 μM), and CGP55845A (1 μM). The internal solution contained the following (in mM): Cs-gluconate 130, CsCl 10, EGTA 0.2, Mg ATP 4, Tri-GTP 0.3, HEPES 10, and QX-314, 4 The pH was adjusted to 7.4 with CsOH, and the osmolarity was 290 mOsm. In all instances, the recordings of spontaneous GABAergic IPSCs usually began at least 5 min after a whole-cell configuration was established with a stable baseline. Spontaneous IPSCs were completely blocked with bath-applied bicuculline methiodide (BMI, 20 μM), confirming that they are mediated by GABA_A receptors. Tonic currents were isolated after bath application of the GABA_A receptor antagonist picrotoxin (100 μM).
Data were recorded with a MultiClamp 700B amplifier, filtered at 10 kHz, and digitized at 20 kHz through a Digidata 1440 interface controlled by pClamp10.7 software (Molecular Devices, CA). Both the frequencies and amplitudes of CA1 sIPSCs were analyzed using Clampfit 10.7 software, and the threshold for detecting sIPSCs was used and followed by visual inspection to ensure the accuracy of detection.

Wang Z., Li Q., Kolls B.J., Mace B., Yu S., Li X., Liu W., Chaparro E., Shen Y., Dang L., del Águila Á., Bernstock J.D., Johnson K.R., Yao J., Wetsel W.C., Moore S.D., Turner D.A, & Yang W. (2023). Sustained overexpression of spliced X-box-binding protein-1 in neurons leads to spontaneous seizures and sudden death in mice. Communications Biology, 6, 252.

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Publication 2023

6,7-dinitroquinoxaline-2,3-dione Amino Acids ARID1A protein, human Bath bicuculline methiodide Capillaries Cells cesium chloride Egtazic Acid GABA-A Receptor GABA-A Receptor Antagonists gluconate HEPES Induced Pluripotent Stem Cells Inhibitory Postsynaptic Currents Medical Devices Microscopy Microscopy, Differential Interference Contrast Neurons Osmolarity Picrotoxin Pyramidal Cells QX-314 Submersion

Whole-cell Patch-clamp Recordings of PVN and DMV Neurons

Whole-cell patch-clamp recordings were performed on visualized PVN and DMV neurons using an infrared-differential interference contrast (IR/DIC) microscope (BX51WI, Olympus, Japan) with a 40x water-immersion objective. Patch pipettes (3–5 MΩ) were pulled from borosilicate glass capillaries (VitalSense Scientific Instruments Co., Ltd) using a four-stage horizontal micropipette puller (P1000, Sutter Instruments, USA), patch pipettes were filled with intracellular solution (solution components in the Supplementary Data 1) were used for voltage-clamp recording. Signals were amplified with a Multiclamp 700B amplifier, low-pass filtered at 2.8 kHz, digitized at 10 kHz, and recorded in a computer for offline analysis using Clampfit 10.7 software (Molecular Devices) (Zhou et al., 2022 (link)).
The current-evoked firing of PVN^CRH neurons was recorded in current-clamp mode (I = 0 pA). The threshold current of the action potential was defined as the minimum current to elicit an action potential. To visualize the PVN neurons, we injected rAAV-DIO-EYFP into the CRH-Cre mice so that green fluorescent EYFP was expressed only in the CRH neurons.
For validation of chemogenetic virus function. After 3 weeks of chemogenetic virus expression, electrophysiological brain slices were prepared by the above process. The PVN neurons expressing m-Cherry were visualized by using a vertical microscope in Mercury lamp mode, and neuronal responses were recorded before and after CNO administration.
In the vitro electrophysiological recordings of light-evoked response, brain slices were prepared by the above process after 3 weeks of optogenetic virus expression, blue light was delivered through an optical fiber (diameter of 200 μm, Inper) that was positioned 0.2 mm above the surface of the target areas. To characterize the function of rAAV-DIO-ChR2-EYFP in the PVN, ChR2-EYFP⁺ neurons in PVN were visualized by a vertical microscope in Mercury lamp mode, and the responses elicited by different frequencies of blue light stimulation (473 nm, 5–8 mV, pulse width 10 Mm, stimulation frequencies 5 Hz, 10 Hz, 20 Hz) were recorded. For recording light-evoked postsynaptic currents (Fang et al., 2020 (link); Zhou et al., 2022 (link)), DMV expressing ChR2-EYFP⁺ fibers were visualized by a vertical microscope in Mercury lamp mode. The membrane potentials were held at −70 mV for recording the excitatory postsynaptic currents and at 0 mV for recording inhibitory postsynaptic currents, and these recordings were immediately terminated once the series resistance changed more than 10%. To eliminate the polysynaptic components, tetrodotoxin (TTX; 1 μM, Dalian Refine Biochemical Items Co., Ltd.) and 4-aminopyridine (4-AP; 2 mM, Sigma) were added to the standard ACSF to block sodium channels and augment light-induced postsynaptic currents, respectively.

Wang X.Y., Chen X.Q., Wang G.Q., Cai R.L., Wang H., Wang H.T., Peng X.Q., Zhang M.T., Huang S, & Shen G.M. (2023). A neural circuit for gastric motility disorders driven by gastric dilation in mice. Frontiers in Neuroscience, 17, 1069198.

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Publication 2023

Action Potentials ARID1A protein, human Brain Capillaries Cardiac Arrest Excitatory Postsynaptic Currents Immersion Inhibitory Postsynaptic Currents Light Medical Devices Membrane Potentials Mercury Mice, Laboratory Microscopy Neurons Optogenetics Photic Stimulation Postsynaptic Current Protoplasm Prunus cerasus Pulse Rate Sodium Channel Virus Virus Physiological Phenomena

Optogenetic Characterization of AAV Constructs

To verify functional characteristics of AAV-DIO-ChR2-mCherry and AAV-DIO-eNpHR3.0-EYFP, pulsed blue light (473 nm, 10 mW, 10 ms pulses) with 5-Hz, 10-Hz, and 20-Hz stimulation protocols, and sustained yellow light (594 nm, 10 mW, 100 ms) were delivered using a laser (Fiblaser, Shanghai) through an optical fiber, respectively. Light-evoked inhibitory postsynaptic currents (IPSCs) were recorded at 0 mV with a blue light (10 mW, 10 ms) in the presence of 4-AP (1 mM) and TTX (1 µM).

Wang H., Liu W.J., Wang X.Y., Chen X.Q., Cai R.L., Zhang M.T., Wang H.T., He G.W., Zhang Z, & Shen G.M. (2023). A central amygdala input to the dorsal vagal complex controls gastric motility in mice under restraint stress. Frontiers in Physiology, 14, 1074979.

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Publication 2023

AP 10 Inhibitory Postsynaptic Currents Light Pulses

Optogenetically Evoked Inhibitory Currents in Nucleus Accumbens

Whole-cell patch-clamp recordings were obtained in the medial shell of the NAc (NAcSh). Data were collected with a Multiclamp 700B amplifier, Digidata 1550B (Molecular Devices, US), and using Clampex 11 (pClamp; Molecular Devices, US). All recordings were acquired in voltage clamp at 34° Celsius and were digitized at 10 kHz and low pass filtered at 2 kHz. Patch pipette was filled with internal solution containing (in mM): 143 CsCl, 10 HEPES, 0.25 EGTA, 5 Phosphocreatine, 4 MgATP, 0.3 NaGTP (295–305 mOsm, pH 7.4 with CsOH) and 1 mg/ml Neurobiotin (cat # SP-1120, Vector Labs, US). CNQX (50 μM, cat # 0190, Tocris Biosciences, UK) and AP5 (10 µM, cat # 0106, Tocris Biosciences, UK) were added to aCSF to block glutamate receptors. All pipettes (3–4 MΩ) were pulled from borosilicate glass (cat # PC-100, Narishige, US). Series resistance (Rs) was monitored throughout the recording for patch sealing. Once whole-cell configuration was obtained, holding potential was set at −70 mV and a 1 ms, 5 mW light pulse was delivered every 10 s (0.1 Hz) through a 40× objective. Once current response was obtained, picrotoxin (100 µM, cat # P1675, Sigma-Aldrich, US) was washed in the recording chamber to verify that optically evoked inhibitory postsynaptic current (oIPSC) was mediated by GABAa receptor activation. Once recording was complete slices were transferred to 4% PFA overnight at 4° Celsius then to 0.1 M PB for post hoc processing of Neurobiotin (cat # SP-1120, Vector Labs, US)^{51 (link)}.

Birnie M.T., Short A.K., de Carvalho G.B., Taniguchi L., Gunn B.G., Pham A.L., Itoga C.A., Xu X., Chen L.Y., Mahler S.V., Chen Y, & Baram T.Z. (2023). Stress-induced plasticity of a CRH/GABA projection disrupts reward behaviors in mice. Nature Communications, 14, 1088.

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Publication 2023

6-Cyano-7-nitroquinoxaline-2,3-dione Adenosine Triphosphate, Magnesium Salt Cardiac Arrest Cells cesium chloride Cloning Vectors Egtazic Acid GABA-A Receptor Glutamate Receptor HEPES Inhibitory Postsynaptic Currents Light Medical Devices neurobiotin Phosphocreatine Picrotoxin Pulse Rate

Electrophysiological Recording of Miniature Inhibitory Postsynaptic Currents

For electrophysiological recordings, 350-µm-thick brain slices were prepared from 3 week old mice and equilibrated in artificial cerebrospinal fluid (aCSF) composed of 120 mM NaCl, 3 mM KCl, 1.3 mM Mg₂SO₄, 1.25 mM NaH₂PO₄, 2.5 mM CaCl₂, 10 mM D-glucose, and 25 mM NaHCO3, and gassed with 95% O₂/5% CO₂, pH 7.3 at room temperature for at least 1 h as described previously [34 (link)].
Patch clamp recordings were performed on coronal slices that were placed in a submerged recording chamber mounted on an upright microscope (BX51WI, Olympus, Hamburg, Germany). Slices were continuously superfused with gassed aCSF (2–3 mL/min, 32 °C, pH 7.3). Recordings of miniature inhibitory postsynaptic current (mIPSC) kinetics were performed using a CsCl-based intracellular solution containing 122 mM CsCl, 8 mM NaCl, 0.2 mM MgCl₂, 10 mM HEPES, 2 mM EGTA, 2 MM Mg-ATP, 0.5 mM Na-GTP, 10 mM QX-314 [N-(2,6-dimethylphenylcarbamoylmethyl) triethylammonium bromide], pH adjusted to 7.3 with CsOH. DL-AP5 (30 μM), CNQX (10 μM) and tetrodotoxin (0.5 μM) were added to the perfusate. mIPSCs were recorded at a holding potential of −70 mV for at least 5 min in aCSF. Data analysis was performed off-line with the detection threshold levels set to 5 pA. The following parameters were determined: frequency, peak amplitude, rise time, time constant of decay (τ-decay), half-width, and electrical charge transfer.

Chen J., Salveridou E., Liebmann L., Sundaram S.M., Doycheva D., Markova B., Hübner C.A., Boelen A., Visser W.E., Heuer H, & Mayerl S. (2023). Triac Treatment Prevents Neurodevelopmental and Locomotor Impairments in Thyroid Hormone Transporter Mct8/Oatp1c1 Deficient Mice. International Journal of Molecular Sciences, 24(4), 3452.

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Publication 2023

6-Cyano-7-nitroquinoxaline-2,3-dione Bicarbonate, Sodium Brain Bromides Cerebrospinal Fluid cesium chloride Egtazic Acid Electricity Glucose HEPES Inhibitory Postsynaptic Currents Kinetics Magnesium Chloride Mice, Laboratory Microscopy Protoplasm QX-314 Sodium Chloride Tetrodotoxin

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Pclamp 9 by Molecular Devices

Sourced in United States, France, Canada

PClamp 9 software is a comprehensive data acquisition and analysis software designed for electrophysiology research. It provides a suite of tools for recording, visualizing, and analyzing electrical signals from cells and tissues.

Picrotoxin by Merck Group

Sourced in United States, United Kingdom, Germany, France, Sao Tome and Principe, Canada, Israel, Australia, Italy, Japan

Picrotoxin is a chemical compound that acts as a GABA antagonist. It is primarily used in scientific research as a tool to study the function of GABA receptors.

Igor pro by Wavemetrics

Sourced in United States, Germany, Jamaica, United Kingdom, Australia

Igor Pro is a graphical data analysis and visualization software developed by WaveMetrics. It is a powerful tool for scientific data processing, analysis, and presentation. Igor Pro provides a wide range of features for data acquisition, processing, and visualization, catering to the needs of researchers, scientists, and engineers across various fields.

More about "Inhibitory Postsynaptic Currents"

Inhibitory Postsynaptic Currents (IPSCs) are the electrical signals generated when inhibitory receptors are activated at the postsynaptic membrane.
These currents play a crucial role in regulating neural network activity and information processing in the brain by reducing the excitability of the postsynaptic neuron.
Studying IPSCs is essential for understanding the balance between excitation and inhibition, which is critical for normal brain function.
Researchers can leverage the power of AI-driven platforms like PubCompare.ai to optimize their IPSC research.
This innovative tool helps locate the best experimental protocols from literature, preprints, and patents, using intelligent comparisons to enhance reproducibility and accuracy.
By harnessing the power of AI, researchers can take their IPSC findings to new heights and unlock new discoveries in the field of neuroscience.
Key techniques and tools used in IPSC research include the Multiclamp 700B amplifier, PClamp 10 software, Clampfit 10, and PClamp software.
These tools enable precise electrophysiological recordings and analysis of IPSC data.
Additionally, the BX51WI microscope and Axopatch 200B amplifier are commonly used in IPSC experiments.
Pharmacological agents like picrotoxin can also be employed to modulate IPSC activity and study its impact on neuronal function.
By combining the latest AI-driven platforms, cutting-edge electrophysiological tools, and a deep understanding of IPSC mechanisms, researchers can push the boundaries of neuroscience research and uncover new insights into the role of inhibitory signaling in brain processes.
With this comprehensive approach, you can enhance the reproducibility, accuracy, and impact of your IPSC research.