Biomedical subjects

O Moran

Publications and source records attributed to O Moran.

At least 55 records · Page 3Linked to original sources

PubMed1992

Level of expression controls modes of gating of a K+ channel.

Several distinct subfamilies of K+ channel genes have been discovered by molecular cloning, however, in some cases the structural differences among them do not account for the diversity of K+ current types, ranging from transient A-type to slowly inactivating delayed rectifier-type, as members within each subfamily have been shown to code for K+ channels of different inactivation kinetics and pharmacological properties. We show that a single K+ channel cDNA of the Shaker subfamily (ShH4) can express in Xenopus oocytes not only a transient A-type K+ current but also, upon increased level of expression, slowly inactivating K+ currents with markedly reduced sensitivity to tetraethylammonium. In correlation with the macroscopic currents there are single-channel gating modes ranging from the fast-inactivation mode which underlies the transient A-type current, to slow-inactivation modes characterized by bursts of longer openings, and corresponding to the slowly inactivating macroscopic currents.

Animals↗

PubMed1992

Ionic permeability of the mitochondrial outer membrane.

The ionic permeability of the outer mitochondrial membrane (OMM) was studied with the patch clamp technique. Electrical recording of intact mitochondria (hence of the outer membrane (OM], derived from mouse liver, showed the presence of currents corresponding to low conductances (less than 50 pS), as well as of four distinct conductances of 99 pS, 152 pS, 220 pS and 307 pS (in 150 mM KCl). The latter were voltage gated, being open preferentially at positive (pipette) potentials. Very similar currents were found by patch clamping liposomes containing the isolated OM derived from rat brain mitochondria. Here a conductance of approximately 530 pS, resembling in its electrical characteristics a conductance already attributed to mitochondrial contact sites (Moran et al. 1990), was also detected. Immunoblot assays of mitochondria and of the isolated OM with antibodies against the outer membrane voltage-dependent anion channel (VDAC) (Colombini 1979), showed the presence of the anion channel in each case. However, the typical electrical behaviour displayed by such a channel in planar bilayers could not be detected under our experimental conditions. From this study, the permeability of the OMM appears different from what has been reported hitherto, yet is more in line with that multifarious and dynamic structure which apparently should belong to it, at least within the framework of mitochondrial biogenesis (Pfanner and Neupert 1990).

Animals↗

PubMed1992

High-conductance pathways in mitochondrial membranes.

The outer and inner membranes of mitochondria have recently been studied with the patch clamp technique. What has emerged is still an ill-defined picture for either membrane, primarily for the wide range of conductances found. Interestingly, however, a few conductances (in the range of 10-80 pS) seem to be ubiquitously distributed. Parallel studies in situ and in reconstituted systems have allowed the assignment to distinct membrane locations of some conductances, whose physiological role is, however, not yet elucidated.

Animals↗

PubMed1992

Physical characterization of the pore forming cytolysine from Gardnerella vaginalis.

The cytolytic toxin (CTox) produced by Gardnerella vaginalis is able to form voltage-dependent cationic channels when incorporated in lipid membranes (Moran et al. (1991) FEBS Lett. 283, 317-320). Osmotic protection experiments show that toxin incorporated in human erythrocytes forms pores between 18 A and 28 A in diameter. A hypothesis of pore formation as a primary event to produce cytolysis is proposed. The CTox activity increases when cells are depolarized by increasing the extracellular K+ concentration, probably reflecting the voltage dependent character of CTox formed channels. The cytolytic effect of the toxin was prevented by low temperatures and was a function of the extracellular Ca2+ concentration, suggesting a Ca2+ influx as part of the lytic mechanism. Binding of CTox to erythrocytes was dependent on external Ca2+ and was less temperature-dependent. Dose-response analysis suggests cooperativity of the toxin for the lytic activity, although no direct evidence of oligomerization has been found.

Blotting, Western↗

PubMed1992

Recurrence rate after fluid aspiration from sonographically benign-appearing ovarian cysts.

The outcome after fluid aspiration from 41 sonographically benign-appearing ovarian cysts was assessed. The considerable probability of recurrence was significantly higher after ultrasound-guided aspiration than following aspiration via laparoscopy (54% versus 30%, respectively, at 36 months). Although all the recurring cysts were benign, the concern with malignancy and the high recurrence rate seem to indicate that fluid aspiration from sonographically benign-appearing cysts by either method is not the management of choice.

Adolescent↗

PubMed1991

N-methyl-D-aspartate receptor-mediated spontaneous activity in cerebellar granule cells in culture.

An outside-out configuration of the patch-clamp method was used to study the properties of spontaneous and glutamate-evoked single channel activity in cerebellar granule cells in culture. Both spontaneous occurring events and glutamate-evoked single channel currents had similar conductances (17.7, 36.5; 17.3, 35.8 pS) and mean open times (0.7, 7.1; 0.6, 7.8 ms). In magnesium free solution, the spontaneous channels were blocked by the NMDA-receptor antagonist APV (0.1 mM) and by the NMDA-channel blocker MK-80 (0.1 mM). The spontaneous activity disappeared at negative holding potentials when the bathing solution contained magnesium (5 mM). Maximum conductance of spontaneous channels increased with days in culture (DIC) from 9 pS at 1 DIC to 59 pS at 10 DIC. These experiments suggest that endogenous transmitter activates NMDA-receptors in cerebellar granule cells in culture.

2-Amino-5-phosphonovalerate↗

PubMed1991

Voltage-dependent cationic channels formed by a cytolytic toxin produced by Gardnerella vaginalis.

A cytolytic toxin produced by G. vaginalis was incorporated in artificial membranes and giant liposomes. The toxin formed ionic channels when incorporated in lipid bilayers. The electrical properties of such channels were studied. Current records revealed a unitary conductance of 126 pS (in symmetrical 150 mM KCl). The open state probability of the cytolysin formed channels was a function of the applied membrane potential. The permeability ratio of cations to anions was estimated to be 6.5.

Cytotoxins↗

PubMed1991

Modulation of a Shaker potassium A-channel by protein kinase C activation.

Brain fast transient K+ channel (A channel) is known to be modulated by PKC activation. We studied, by two-electrode voltage clamp, the molecular mechanism of modulation by PKC activation of A-channels expressed in Xenopus oocytes from the Shaker H4 clone. The modulation is inhibitory affecting primarily the maximal conductance of the channels. A secondary effect is a small change in the voltage-dependence of activation and inactivation of the channel.

Animals↗

PubMed1991

Competitive blockage of the sodium channel by intracellular magnesium ions in central mammalian neurones.

The aim of this study was to determine from macroscopic current analysis how intracellular magnesium ions, Mgi2+, interfere with sodium channels of mammalian neurones. It is reported here that permeation across the sodium channel is voltage- and concentration-dependently reduced by Mgi2+. This results in a general reduction of sodium membrane conductance and an outward sodium peak current at large positive potentials. 30 mM Mgi2+ leads ot a negative shift of voltage dependence of sodium channel gating parameters, probably due to the surface potential change of the membrane. This shift alone is, however, insufficient to explain the reduction of outward sodium currents. The blockage by Mgi2+ is decreased upon increasing intracellular or extracellular Na+ concentration, which suggests that Mgi2+ interferes with sodium permeation by competitively occupying sodium channels. Using a kinetic model to describe the sodium permeation, the dissociation constant (at zero membrane potential) of Mgi2+ for the sodium channel has been calculated to be 8.65 +/- 1.51 mM, with its binding site located at 0.26 +/- 0.05 electrical distance from the inner membrane. This dissociation constant is smaller than that of Nai+, which is 83.76 +/- 7.60 mM with its binding site located at 0.75 +/- 0.23. The low dissociation constant of Mgi2+ reflects its high affinity for the sodium channel.

Animals↗

PubMed1991

Voltage dependent calcium channels in cerebellar granule cell primary cultures.

Voltage activated calcium channels were studied in rat cerebellar granule cells in primary culture. Macroscopic currents, carried by 20mM Ba2+, were measured in the whole-cell configuration. Slowly inactivating macroscopic currents, with a maximum value at a membrane potential around 5 mV, were recorded between the 1st and the 4th day in culture. These currents were completely blocked by 5mM Co2+, partially blocked by 10 microM nifedipine, and increased by 2 to 5 microM BAY K-8644. Two types of channels, in the presence of 80 mM Ba2+, were identified by single channel recording in cell-attached patches. The first type, which was dihydropyridine agonist sensitive, had a conductance of 18 pS, a half activation potential of more than 10 mV and did not inactivate. This type of channel was the only type found during the first four days in culture, although it was also present up to the 11th day. The second type of channel was dihydropyridine insensitive, had a conductance of 10 pS, a half activation potential less than -15 mV, and displayed voltage dependent inactivation. This second type of channel was found in cells for more than four days in culture.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

PubMed1991

Curettage vs. nonsurgical management in women with early spontaneous abortions. The effect on fertility.

Among 114 women with early spontaneous abortions, low beta-human chorionic gonadotropin levels and no ultrasonographically demonstrable residual tissue in the uterine cavity, 68 underwent uterine curettage, and 46 were managed conservatively. No complications, such as bleeding or infection, were observed in either group, and the probability of conception, outcome of subsequent pregnancies and rate of subsequent abortions were similar. Curettage offers no advantage over conservative management in selected patients with early abortions.

Abortion, Spontaneous↗

PubMed1990

Voltage-dependent calcium currents in trigeminal chick neurons.

The presence of action potentials, when sodium and potassium currents were blocked, has been investigated in trigeminal ganglion neurons, using the patch-clamp technique. In this conditions, inward currents, sensitive to the external application of cadmium, were detected. Activation and inactivation properties were investigated, as well as the behaviour of the current in the presence of extracellular Barium. The properties of these inward currents in trigeminal neurons are correlated to high threshold voltage-dependent calcium channels.

Action Potentials↗

PubMed1990

Electrophysiological characterization of contact sites in brain mitochondria.

From morphological and biochemical studies it has been recognized that the regions where the outer and inner membranes of mitochondria come in close contact (contact sites) can be the route mechanism through which mitochondria interact directly with the cytoplasm. We have studied these regions electrophysiologically with the patch clamp technique, with the aim of understanding if this direct interaction is mediated by high conductance ion channels similar to the channel already detected in the inner membrane of mitochondria (Sorgato M. C., Keller, B. U., and Stühmer, W. (1987) Nature 330, 498-500). Contact sites isolated from rat brain mitochondria were thus incorporated into liposomes subsequently enlarged sufficiently to be patch clamped. This study shows that these particular fractions contain ion channels with conductances ranging from approximately 5 picosiemens to 1 nanosiemens (in symmetrical 150 mM KCl). Most of these channels are not voltage-dependent and can be open at physiological potentials sustained by respiring mitochondria. The lack of voltage sensitivity seems not to be the outcome of methodological artifacts, as voltage-gated channels are detected in giant liposomes containing either the outer mitochondrial membrane or a partially purified fraction of the inner mitochondrial membrane. These data therefore indicate that channels present in mitochondrial contact sites have properties which render them amenable to perform several of the functions hypothesized for these regions, particularly that of translocating macromolecules from the cytoplasm to the matrix of mitochondria.

Animals↗

PubMed1990

Kainate activated single channel currents as revealed by domoic acid.

We have studied the properties of kainic acid receptor-activated channels using domoic acid as an agonist. Similarities of the electrophysiological, pharmacological and noise properties of domoic acid and kainic acid-evoked currents confirm that domoate is a potent and specific agonist of the kainate receptor. Single-channel properties of domoic acid-evoked currents were directly determined from outside-out membrane patches for the first time, and results were compared with those obtained by fluctuation analysis of macroscopic currents. Small conductance cationic-selective channels of approximately 4 pS and a mean open time of 2 to 3 ms were detected using both methods.

Animals↗

PubMed1990

Sodium ionic and gating currents in mammalian cells.

Ionic and gating currents from voltage-gated sodium channels were recorded in mouse neuroblastoma cells using the path-clamp technique. Displacement currents were measured from whole-cell recordings. The gating charge displaced during step depolarizations increased with the applied membrane potential and reached saturating levels above 20 mV. Prolonged large depolarizations produced partial immobilization of the gating charge, and only about one third of the displaced charge was quickly reversed upon return to negative holding potentials. The activation and inactivation properties of macroscopic sodium currents were characterized by voltage-clamp analysis of large outside-out patches and the single-channel conductance was estimated from non-stationary noise analysis. The general properties of the sodium channels in mouse neuroblastoma cells are very similar to those previously reported for various preparations of invertebrate and vertebrate nerve cells.

Animals↗

PubMed1990

Short- and long-term desensitization of serotonergic response in Xenopus oocytes injected with brain RNA: roles for inositol 1,4,5-trisphosphate and protein kinase C.

In Xenopus oocytes injected with rat brain RNA, serotonin (5HT) and acetylcholine (ACh) evoke membrane responses through a common biochemical cascade that includes activation of phospholipase C, production of inositol 1,4,5-trisphosphate (Ins1,4,5-P3), release of Ca2+ from intracellular stores, and opening of Ca-dependent Cl- channels. The response is a Cl- current composed of a transient component (5HT1 or ACh1) and a slow, long-lasting component (5HT2 or ACh2). Here we show that only the fast, but not the slow, component of the response is subject to desensitization that follows a previous application of the transmitter. The recovery of 5HT1 from desensitization is biphasic, suggesting the existence of two types of desensitization: short-term desensitization (STD), which lasts for less than 0.5 h; and long-term desensitization (LTD) lasting for up to 4 h. The desensitization between 5HT and ACh is heterologous and long-lasting. We searched for (a) the molecular target and (b) the cause of desensitization. (a) Pre-exposure to 5HT does not reduce the response evoked by intracellular injection of Ca2+ and by Ca2+ influx. Cl- current evoked by intracellular injection of Ins1,4,5-P3 was reduced shortly after application of 5HT, but fully recovered 30 min later. Thus, the Cl- channel is not a target for desensitization. Neither Ins1,4,5-P3 receptor nor the Ca2+ store is a target of LTD but they may be the targets of STD. (b) Ca2+ injection did not inhibit the 5HT response, suggesting that Ca2+ is not a sole cause of STD or LTD.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗

PubMed1990

Voltage dependent sodium currents in cultured rat cerebellar granule cells.

Sodium currents were studied in granule cells dissociated from rat cerebellum. Macroscopic currents were recorded using the patch-clamp technique. Sodium currents, which are TTX sensitive, reached a maximum peak value of 0.42 +/- 0.08 pA/microns2 at 18.4 +/- 2.2 mV (n = 6). Activation and inactivation kinetics and steady-state properties were described in terms of Hodgkin and Huxley parameters. The properties of sodium channels in cultured rat cerebellar granule cells are very similar to those reported for various neural preparations.

Action Potentials↗

PubMed1990

Prediction of residual trophoblastic tissue in first-trimester abortions and low levels of human chorionic gonadotropin beta-subunit.

The clinical data and the histologic report of curettage specimens of 174 patients with first-trimester abortion and human chorionic gonadotropin beta-subunit less than 500 mIU/ml were reviewed. In only 22 (12.6%) were chorionic villi histologically reported. A multivariate analysis showed that increased log human chorionic gonadotropin beta-subunit levels within the range of 20 to 500 mIU/ml, intrauterine tissue demonstrated by ultrasonography, higher gestational age, and active bleeding were significant predictive variables for the presence of villi in curettage specimens. A mathematical formula and a clinical scoring system for the prediction of residual villi in these cases were devised and were applied to the data of our study group. The sensitivity, false-negative rate, and specificity of the scoring system were 90.9%, 2.2%, and 59.2%, respectively; however, the false-positive rate was 75.6%. By use of this scoring system in patients with first-trimester abortion and low human chorionic gonadotropin beta-subunit level, the presence of villi in curettage specimens can thus be predicted with a high sensitivity and as many as 60% of curettage procedures might be prevented.

Abortion, Induced↗