No abstract available.
Cochlear Nucleus ; Patch-Clamp Techniques

Brain ; N-Methylaspartate ; Neurons ; Patch-Clamp Techniques

Action Potentials ; Brain ; Neurons ; Patch-Clamp Techniques

A USB bus based data interface technology for full auto Patch-Clamp system is discussed in the article. The main controller is CY2131QC (Cypress) and the logic controller is EPM3256A (Altera). Optocouplers are used to get rid of the noise from the interface. It makes the installation of the Patch-Clamp system easier by using the USB bus, and is suitable for the new generation of the Patch-Clamp system with a high speed of 1M bytes/s.
Computer-Aided Design ; Equipment Design ; Patch-Clamp Techniques ; instrumentation ; methods

OBJECTIVECurrents contributing repolarization in rabbit ventricular myocyte are very complex since the I(To.s) covers almost the whole repolarization phase of the action potential. The other components of repolarizing currents, as I(Kr) and I(Ks) are small. The purpose of this study is to investigate whether or not there are other currents in rabbit ventricular repolarization.

METHODSIon currents of rabbit ventricular myocyte were recorded using the whole-cell patch-clamp technique.

RESULTSIn the present work, an nonselective cation current was identified by replacing the K(+) with Cs(+) in the bathing and pipette solutions. The outward current elicited by depolarizing potentials could be inhibited by Gd(3+), an effective inhibitor of nonselective cation currents. Depleting Ca(2+) and Mg(2+) in the bathing solution, the amplitudes of this outward current increased by 40%-116% at +60 mV, and adding 2 micromol/L insulin to the solution (with normal concentration of Ca(2+) and Mg(2+) in Tyrode's solution), the amplitude increased by 30%-60% at +60 mV.

CONCLUSIONIt is suggested that a nonselective cation current in rabbit ventricular myocytes may play an important role in the repolarization of the action potential in rabbit ventricle. Changes of nonselective cation current will lead to induce or inhibit arrhythmia.

In this paper we present an easily available method of intracellular dialysis via a microcatheter inserted into glass pipette during patch clamp experiment. An oblique hole through the glass pipette holder (above the lateral hole for cell-seal suction) is drilled, through which a microcatheter (O.D.=0.1 mm) made from the universal pipetter tip by hand-drawing passes and sticks out of the holder mouth in parallel with the Ag-AgCl electrode. With a syringe connected to the microcatheter, substitution of intracellular solution and intracellular dialysis of drugs can be achieved easily. Compared with repatch technique and intracellular solution substitution techniques used abroad, this method operates more easily and can produce more reliable results.
Dialysis ; instrumentation ; methods ; Equipment Design ; Microelectrodes ; Patch-Clamp Techniques ; instrumentation

Patch clamp is a technique that can measure weak current in the level of picoampere (pA). It has been widely used for cellular electrophysiological recording in fundamental medical researches, such as membrane potential and ion channel currents recording, etc. In order to obtain accurate measurement results, both the resistance and capacitance of the pipette are required to be compensated. Capacitance compensations are composed of slow and fast capacitance compensation. The slow compensation is determined by the lipid bilayer of cell membrane, and its magnitude usually ranges from a few picofarads (pF) to a few microfarads (μF), depending on the cell size. The fast capacitance is formed by the distributed capacitance of the glass pipette, wires and solution, mostly ranging in a few picofarads. After the pipette sucks the cells in the solution, the positions of the glass pipette and wire have been determined, and only taking once compensation for slow and fast capacitance will meet the recording requirements. However, when the study needs to deal with the temperature characteristics, it is still necessary to make a recognition on the temperature characteristic of the capacitance. We found that the time constant of fast capacitance discharge changed with increasing temperature of bath solution when we studied the photothermal effect on cell membrane by patch clamp. Based on this phenomenon, we proposed an equivalent circuit to calculate the temperature-dependent parameters. Experimental results showed that the fast capacitance increased in a positive rate of 0.04 pF/℃, while the pipette resistance decreased. The fine data analysis demonstrated that the temperature rises of bath solution determined the kinetics of the fast capacitance mainly by changing the inner solution resistance of the glass pipette. This result will provide a good reference for the fine temperature characteristic study related to cellular electrophysiology based on patch clamp technique.
Cell Membrane ; Electric Capacitance ; Membrane Potentials ; Patch-Clamp Techniques ; Temperature

The inferior colliculus (IC) is a pivot along the central auditory pathway. Using infrared visual whole-cell patch clamp recording technique, we investigated the electrophysiological properties of IC subnuclei neurons. Recordings were made from 88 neurons, including 21 neurons from the dorsal cortex of the IC (ICd), 43 neurons from the central nucleus of the IC (ICc) and 24 neurons from the external cortex of the IC (ICx). Based on the responses to positive current injection, three firing patterns, i.e., onset (6.8%, n = 6), adapting (39.8%, n = 35) and sustained (53.4%, n = 47) patterns, were identified. The hyperpolarization-activated inward current (Ih) could be recorded in half of the neurons (49/88). The sustained pattern occurred in more than half of ICd and ICc neurons (61.9% and 67.4%), while the adapting pattern occurred in majority of ICx neurons (75%). Action potential (AP) threshold and time constant also showed significant differences across neurons from the ICd, the ICc and the ICx. Our results indicate that IC neurons are different in electrophysiological properties across the subnuclei. The variance of the responses may be related to the distinct types of neurons as well as the received projections, which is implicated in the distinct roles of IC neurons in central auditory processing.
Action Potentials ; Animals ; Electrophysiological Phenomena ; In Vitro Techniques ; Inferior Colliculi ; cytology ; Mice ; Neurons ; cytology ; Patch-Clamp Techniques

BACKGROUND: Human neural stem cells have at least two types of voltage-dependent potassium channels including inward and outward rectifying potassium channels. Data for the effects of propofol, intravenous anesthetics, on human neural potassium channels are limited. We have examined the effects of propofol on voltage-dependent potassium channels in human neural stem cells. METHODS: Voltage-dependent potassium currents were measured in cultured human neural stem cells using the whole cell patch-clamp technique before and after application of 0.1 mM propofol. Inward and outward rectifying potassium currents were elicited by hyperpolarizing and depolarizing step pulses from -60 mV holding potential, respectively. RESULTS: Propofol was found to reversibly block the outward rectifying potassium current (p<0.05), while the anesthetics did not alter the characteristics of inward rectifying potassium current. The blocking effect of propofol on outward rectifying current was voltage-dependent and enhanced at depolarization potentials above +30 mV (p<0.05). Propofol also decreased the slope conductance of outward potassium current (p<0.05). CoNCLUSION: These results show that propofol may strongly affect the outward potassium channels in human neural stem cells and also suggest a need for investigation into the effect of propofol on the functional regulation of human neural stem cells.
Anesthetics ; Anesthetics, Intravenous ; Humans* ; Neural Stem Cells ; Patch-Clamp Techniques ; Potassium Channels* ; Potassium* ; Propofol* ; Stem Cells*

Various voltage-gated K+ currents were recently described in dorsal root ganglion (DRG) neurons. However, the characterization and diversity of voltage-gated K+ currents have not been well studied in trigeminal root ganglion (TRG) neurons, which are similar to the DRG neurons in terms of physiological roles and anatomy. This study was aimed to investigate the characteristics and diversity of voltage-gated K+ currents in acutely isolated TRG neurons of rat using whole cell patch clamp techniques. The first type (type I) had a rapid, transient outward current (I(A)) with the largest current size having a slow inactivation rate and a sustained delayed rectifier outward current (I(K)) that was small in size having a fast inactivation rate. The I(A) currents of this type were mostly blocked by TEA and 4-AP, K channel blockers whereas the I(K) current was inhibited by TEA but not by 4-AP. The second type had a large I(A) current with a slow inactivation rate and a medium size-sustained delayed IK current with a slow inactivation rate. In this second type (type II), the sensitivities of the I(A) or I(K) current by TEA and 4-AP were similar to those of the type I. The third type (type III) had a medium sized I(A) current with a fast inactivation rate and a large sustained I(K) current with the slow inactivation rate. In type III current, TEA decreased both I(A) and I(K) but 4-AP only blocked I(A) current. The fourth type (type IV) had a smallest I(A) with a fast inactivation rate and a large IK current with a slow inactivation rate. TEA or 4-AP similarly decreased the I(A) but the I(K) was only blocked by 4-AP. These findings suggest that at least four different voltage-gated K+ currents in biophysical and pharmacological properties exist in the TRG neurons of rats.
Animals ; Diagnosis-Related Groups ; Ganglia, Spinal ; Ganglion Cysts ; Neurons ; Patch-Clamp Techniques ; Potassium ; Rats ; Tea