Slowing of ERG current deactivation in NG108-15 cells by the histidine-specific reagent diethylpyrocarbonate.

The aim of this study was to explore and characterize the effect of the histidine-specific reagent diethylpyrocarbonate (DEPC) on the ERG (ether-à-go-go related gene) channels of whole-cell voltage-clamped NG108-15 neuroblastoma x glioma hybrid cells. The channels were fully activated by long depolarizing prepulses. Hyperpolarizing pulses elicited K+ inward currents which deactivated after reaching a peak.

Effect of low external calcium on the ERG current of NG108-15 cells.

K(+) currents through ERG (ether-à-go-go related gene) channels were recorded in whole-cell voltage clamped NG108-15 neuroblastomaxglioma hybrid cells. The channels were fully activated by low holding potential (V(H)=-20 mV) and long depolarizing prepulses. Hyperpolarizing pulses elicited inward currents which deactivated after reaching a peak.

Cell-type specific expression of ATP-sensitive potassium channels in the rat hippocampus.

1. The distribution of ATP-sensitive K+ channels (KATP channels) was investigated in four cell types in hippocampal slices prepared from 10- to 13-day-old rats: CA1 pyramidal cells, interneurones of stratum radiatum in CA1, complex glial cells of the same area and granule cells of the dentate gyrus. The neuronal cell types were identified visually and characterized by the shapes and patterns of their action potentials and by neurobiotin labelling.

[Malaria--biological aspects of an infectious disease of importance to humans].

Malaria is the most prevalent parasite-transmitted infectious disease in humans, with 300-500 million people infected and 3-5 million persons dying from the disease each year [1]. There is, however, surprisingly little knowledge of the parasite's biology and its evolutionary adaptations to cope with a life as an intracellular parasite within its vertebrate host. This article gives an overview of the parasite's developmental cycle and highlights aspects of the immune response in infected humans and the parasite's mechanisms of immune evasion.

Caffeine-evoked contractures in single slow (tonic) muscle fibres of the frog (Rana temporaria and R. esculenta).

Single slow (tonic) muscle fibres were dissected from cruralis muscles of Rana temporaria and R. esculenta. Increasing concentrations of caffeine were applied in Ringer solution, and contractures were measured isometrically.

Long-term potentiation and functional synapse induction in developing hippocampus.

Long-term potentiation (LTP) is a cellular mechanism that potentially underlies learning and memory. To test the hypothesis that LTP is involved in activity-dependent synapse formation, we combined whole-cell recordings and confocal microscopy to investigate hippocampal glutamatergic synapses at their earliest stages of development. Here we report that, during the first postnatal week, the hippocampal glutamatergic network becomes gradually functional owing to the transformation of precursor, pure NMDA (N-methyl-D-aspartate)-receptor-based synaptic contacts into conducting AMPA (alpha-amino-3-hydroxy-5-methylisoxazole-4-proprionate)/NMDA-re cep tor-type synapses.

Long-term potentiation as a mechanism of functional synapse induction in the developing hippocampus.

During the first 2 days of postnatal development, CA1 hippocampal glutamatergic synaptic transmission is based almost exclusively on NMDA receptors and is non-functional at resting potential. Within the following days an increasing number of functionally mature synapses, containing both NMDA and AMPA receptors, were observed. We found that the maturation of the NMDA receptor-mediated synapses could be induced experimentally with a pairing protocol, a process termed functional synapse induction.

Model experiments on squid axons and NG108-15 mouse neuroblastoma x rat glioma hybrid cells.

Three types of ionic current essentially determine the firing pattern of nerve cells: the persistent Na+ current, the M current and the low-voltage-activated Ca(2)+ current. The present article summarizes recent experiments concerned with the basic properties of these currents. Keynes and Meves (Proc R Soc Lond B (1993) 253, 61-68) studied the persistent or steady-state Na+ current on dialysed squid axons and measured the probability of channel opening both for the peak and the steady-state Na+ current (PF(peak) and PF(ss)) as a function of voltage.

Inhibitory modulation of fast and slow Ca(2+)-currents in neuroblastoma x glioma cells during differentiation.

Mouse neuroblastoma x rat glioma hybrid cells (N x G, 108CC15) were used to study the inhibitory effects of the synthetic opioid D-Ala2-D-Leu5-enkephalin (DADLE), somatostatin, adrenaline-alpha 2 and angiotensin II on voltage-dependent Ca(2+)-currents (ICa) using the patch-clamp technique in the whole-cell configuration mode. The inhibitory effects could be abolished by pretreatment of N x G cells with pertussis toxin or intracellular infusion of GDP beta S indicating an involvement of a pertussis toxin sensitive GTP-binding protein (G-protein), presumably Go. The effect of DADLE, the strongest inhibitor of ICa, was studied during dibutyryl cyclic AMP (dBcAMP) induced differentiation.

Phloretin affects the fast potassium channels in frog nerve fibres.

The effect of phloretin (20-100 microM), a dipolar organic compound, on the voltage clamp currents of the frog node of Ranvier has been investigated. The Na currents are simply reduced in size but not otherwise affected. Phloretin has no effect on the slow 4-aminopyridine-resistant K channels.

Use-dependent block with tetrodotoxin and saxitoxin at frog Ranvier nodes. II. Extrinsic influence of cations.

Use-dependent declines of Na+ currents in myelinated frog nerve fibres were measured during a train of depolarizing pulses in solutions containing tetrodotoxin (TTX) or saxitoxin (STX). The following effects of external monovalent (Na+), divalent (Ca2+, Mg2+) and trivalent (La3+) cations on use dependence were found: Increasing the Ca2+ concentration from 2 to 8 mM shifts its voltage dependence by 20 mV whereas no significant use-dependent decline occurred at 0.2 mM Ca2+.

Use-dependent block with tetrodotoxin and saxitoxin at frog Ranvier nodes. I. Intrinsic channel and toxin parameters.

The use-dependent phasic blockage of sodium channels by tetrodotoxin (TTX) and saxitoxin (STX) was examined in frog nodes of Ranvier using trains of depolarizing pulses. The decline of the peak Na+ current from its initial value (I0) before the train to a stationary value (I infinity) after the train was more pronounced at more negative holding potentials. The relationship between I infinity/I0 and holding potential was fitted by a sigmoid function which yielded values for the steepness of the voltage dependencies of around -15 mV for TTX and -8 mV for STX.

Interaction of monovalent cations with tetrodotoxin and saxitoxin binding at sodium channels of frog myelinated nerve.

Na currents and Na-current fluctuations were measured in myelinated frog nerve fibres to study interactions between monovalent externally applied cations and the binding of the Na-channel blockers tetrodotoxin (TTX) or saxitoxin (STX). Adding 110 mM NaCl to Ringer's solution increased the maximum peak Na conductance by a factor of 2.51 in the presence of 12 nM TTX and by a factor of 2.

Binding of tetrodotoxin and saxitoxin to Na+ channels at different holding potentials: fluctuation measurements in frog myelinated nerve.

The number of available Na+ channels in nodes of frog nerve fibres was determined from nonstationary Na+ current fluctuations recorded during a train of depolarizing test pulses. Mean numbers in Ringers's solution were 90,000 at a hyperpolarizing holding potential VH = -40 mV, 50,000 at the resting potential (VH = 0 mV) and 30,000 at a depolarizing holding potential VH = 30 mV. Addition of the cationic channel blockers tetrodotoxin (TTX) or saxitoxin (STX) to Ringer reduced the channel number by a factor which was independent of the holding potential.

Use-dependent block of sodium channels in frog myelinated nerve by tetrodotoxin and saxitoxin at negative holding potentials.

Na+ currents were measured in myelinated frog nerve fibres in the presence of nanomolar concentrations of tetrodotoxin (TTX) or saxitoxin (STX) in the extracellular solution. The Na+ currents declined during a train of depolarizing pulses if the fibre was held at hyperpolarizing potentials between the pulses. At a pulse frequency of 0.

ATP-sensitive potassium channels in adult mouse skeletal muscle: different modes of blockage by internal cations, ATP and tolbutamide.

Single ATP-sensitive K channels were studied in membrane patches excised from enzymatically dissociated mouse toe muscle. The channel conductance is 74 pS in symmetrical 160 mM KCl solutions. Replacement of K+ by Na+ in the internal solution or 2 mM internal Ca2+ or Mg2+ induced a rectification of the current-voltage curve at positive potentials.

Properties and calcium-dependent inactivation of calcium currents in cultured mouse pancreatic B-cells.

1. Ca2+ currents were recorded using the whole-cell mode of the patch-clamp technique from mouse pancreatic B-cells kept in culture for 1-4 days. B-cells were identified in the cell-attached mode by their response to a change in the glucose concentration from 3 to 15 or 20 mM or by their inward currents.

Modification of L-type calcium current by intracellularly applied trypsin in guinea-pig ventricular myocytes.

1. The L-type Ca2+ current was recorded in guinea-pig ventricular myocytes by the patch clamp technique in the whole-cell configuration. The modification of the current by intracellular application of proteases was studied.

The effect of D600 on potassium contractures of slow muscle fibres of Rana temporaria.

(1) The effect of 30 microM D600 on the amplitude and time course of isometric contractures was studied in single slow fibres of Rana temporaria. (2) D600 only slightly reduced the amplitude of contractures evoked with 30 or 95 mM K-Ringer's. Maintenance of tension was strongly impaired by D600 only during exposure to 95 mM K.

Effects of a protein phosphatase inhibitor, okadaic acid, on membrane currents of isolated guinea-pig cardiac myocytes.

The effects of a protein phosphatase inhibitor, okadaic acid (OA), were studied on membrane currents of isolated myocytes from guinea-pig cardiac ventricle. The whole-cell Ca2+ current (ICa) was recorded as peak inward current in response to test pulse to 0 mV. Extracellular application of OA (5-100 microM) produced an increase of ICa.

Structural alterations in the membrane of the slow muscle fiber of Rana temporaria after denervation.

Mixed fiber bundles as well as separated slow and fast fibers from normal and denervated muscles of Rana temporaria were freeze-fractured. The membranes of both fiber types are distinguished in this species by the presence of fairly regularly distributed particle aggregates or arrays of different shapes and sizes; the number per unit area of the membrane is six times higher in fast than in slow fibers. The intramembrane particle (IMP) density is higher in slow than in fast fibers.

Na+ currents in cultured mouse pancreatic B-cells.

Pancreatic B-cells, kept in culture for 1-4 days, were studied in the whole-cell, cell-attached and outside-out modes of the patch clamp technique. B-cells were identified by the appearance of electrical activity in the cell-attached mode when the bath glucose was raised from 3 to 20 mM. In whole-cell, 80% of these cells showed a transient inward Na+ current, when depolarizing pulses were preceded by holding potentials, or prepulses to potentials more negative than -80 mV.

"Run-down" of the Ca current during long whole-cell recordings in guinea pig heart cells: role of phosphorylation and intracellular calcium.

We examined by a statistical approach the decrease of the Ca current ("run-down") during long-lasting recordings with the whole-cell patch-clamp technique in guinea pig ventricular myocytes. The results are as follows. (1) Run-down of the Ca current (ICa) occurs in three phases (T1-T3).