Biomedical subjects

M Martina

Publications and source records attributed to M Martina.

8 recordsLinked to original sources

PubMed1995

Spontaneous GABA-mediated synaptic currents in cerebellar granule cells in culture.

The whole-cell configuration of the patch clamp technique was used to characterize the electrophysiological properties of spontaneous GABA-mediated synaptic currents in cerebellar granule cells grown in a low (5 mM) potassium medium. In the presence of kynurenic acid (1 mM), to block the excitatory drive, bicuculline-sensitive synaptic events were recorded. Their amplitude distribution could be fitted by several Gaussians having the same interpeak distance. In tetrodotoxin (TTX, 1 microM) spontaneous miniature events occurred at a lower frequency. Spontaneous currents reversed polarity at 8.17 +/- 0.63 mV, a potential close to ECl; the decay phase could be fitted with a single exponential having at -60 mV a time constant of 48.7 +/- 1.5 ms. In low noise recordings, channel closing could be resolved during the decay phase of miniature events. It appeared that a single quantum of GABA opened few channels on the postsynaptic membrane.

Animals↗

PubMed1995

Whole cell and single channel properties of a new GABA receptor transiently expressed in the Hippocampus.

1. The patch-clamp technique was used to characterize, in acutely dissociated CA3 rat hippocampal neurons, the whole cell and single channel properties of a novel response to gamma-aminobutyric acid (GABA) present only during a restricted period of postnatal development. 2. At postnatal days 0-10 (P0-P10), both GABA (100 microM) and isoguvacine (50 microM) evoked at a holding potential of -50 mV, in symmetrical chloride solution, whole cell inward currents. Bicuculline blocked the response to isoguvacine but only reduced the response to GABA (from 512 +/- 137 pA to 60 +/- 13 pA, mean +/- SE). After P12, bicuculline abolished the response to GABA. 3. The bicuculline-insensitive GABA currents were Cl- mediated and antagonized by picrotoxin. The desensitization rate was slower than the conventional bicuculline-sensitive response. The peak to plateau ratio induced by 0.1 or 1 mM of GABA shifted from 4.6 +/- 0.4 and 17.7 +/- 2.6 to 1.5 +/- 0.1 and 3.1 +/- 0.5 in the absence or in the presence of bicuculline, respectively. The recovery from desensitization was significantly faster for the bicuculline-insensitive responses. 4. In excised outside-out patches, GABA (20 microM) activated, in the presence of bicuculline (100 microM), single channel currents having conductances of 14, 22, and 31 pS. These values were similar to those obtained in the same preparation, in the absence of bicuculline. 5. These findings suggest that this new receptor type, which mediates bicuculline-insensitive responses with slow kinetics, may potentiate the depolarizing action of GABA during a critical period of postnatal development and therefore play a crucial role in synaptogenesis.

Animals↗

PubMed1994

Developmental changes in spontaneous GABAA-mediated synaptic events in rat hippocampal CA3 neurons.

Ongoing spontaneous postsynaptic potentials (SPSPs) were intracellularly recorded at 34-36 degrees C from hippocampal CA3 neurons in slices obtained from postnatal days (P) 0-6 and 7-31. SPSPs occurred randomly, and their frequency distribution was fitted by a single exponential function. They were little affected by kynurenic acid, but were reversibly blocked by bicuculline, implying that they were mediated by GABAA receptors. The mean amplitude was 4.53 +/- 0.89 mV in control conditions and 4.07 +/- 0.79 mV in kynurenic acid. In kynurenic acid (with CsCl-filled microelectrodes), SPSPs reversed polarity at 2.4 +/- 2 mV. When tetrodotoxin (1 microM) was added to kynurenic acid solution, GABAA-mediated miniature postsynaptic potentials (MPSPs) were recorded. Under these conditions large events disappeared. The mean amplitude of MPSPs was 2.51 +/- 0.43 mV. The mean frequency decreased from 2.96 +/- 1.04 Hz in kynurenic acid to 0.4 +/- 0.15 Hz in kynurenic acid plus tetrodotoxin. In contrast with P0-P6, at P7-P31 SPSPs were significantly affected by kynurenic acid. The mean amplitude of SPSPs shifted from 4.71 +/- 0.82 mV in control conditions to 3.79 +/- 0.76 mV in kynurenic acid. At this developmental stage, the reversal potential of GABAA-mediated SPSPs shifted towards more negative values (-23.7 +/- 1.3 mV). Addition of tetrodotoxin to kynurenic acid solution abolished larger events and revealed GABAergic MPSPs. The mean amplitude of MPSPs was 2.72 +/- 0.5 mV, a value very close to that observed at P0-P6. Synaptic currents were recorded at 22-24 degrees C from voltage-clamped CA3 pyramidal neurons (at P6) using the tight-seal whole-cell recording technique.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials↗

PubMed1994

The effect of intracellular Ca2+ on GABA-activated currents in cerebellar granule cells in culture.

The patch clamp technique was used to study the effects of intracellular free calcium ([Ca2+]i) on GABAA-evoked whole-cell and single channel currents of cultured cerebellar granule cells. Changes in [Ca2+]i were obtained by adding to the extracellular solution the calcium ionophore A23187 (2 microM). The relationship between [Ca2+]i and [Ca2+]o in the presence or absence of A23187 was assessed using fluorimetric measurements from Fura-2 loaded cells. In 2 mM [Ca2+]o and A23187, [Ca2+]i was about 1.5 microM, whereas in the absence of A23187 it was about 250 nM. In whole-cell experiments (symmetrical chloride concentrations) at -50 mV, GABA (0.5 microM) evoked inward currents that did not desensitize. Bath application of A23187 significantly reduced the steady-state amplitude of GABA currents by 37 +/- 6%. Single channel currents activated by GABA (0.5 microM) were also recorded in the outside-out configuration of the patch clamp technique. Kinetic analysis of single channel events revealed that A23187 significantly increased the long closed time constant (tau c3) without affecting the open time constants (tau o1 and tau o2) or the short and medium closed time constants (tau c1 and tau c2). Moreover, application of A23187 induced a significant reduction of burst duration (tau b). We conclude that a rise in [Ca2+]i by A23187 may decrease the binding affinity of GABA for the GABAA receptor.

Animals↗

PubMed1992

Calcium-activated potassium channels in chondrocytes.

The presence of calcium-activated potassium channels in chondrocytes of growing cartilage was tested. Results obtained with fura-2 on cultured resting chondrocytes indicate that the cells respond to an elevation of extracellular calcium concentration ([Ca2+]o) from 0.1 to 2 mM increasing the intracellular concentration of the ion ([Ca2+]i) from 117 to 187 nM. This increment may be blocked by 3 microM La3+. Patch clamp experiments in cell-attached configuration showed that, when [Ca2+]i rises, the open probability (Po) of the K+ channels increases. Increments in both Po and unitary currents of the K+ channels can be obtained after applying 2.5 microM A23187 with 2 mM [Ca2+]o. Hence, the results demonstrate that, in chondrocytes, a class of Ca(2+)-activated K+ channels is present and their activity is related to an increase of [Ca2+]i.

Animals↗

PubMed1990

A potassium channel in cultured chondrocytes.

Chondrocytes, obtained from preosseous cartilage, were studied by patch clamp technique in cell-attached recording configuration, and single potassium channels were characterized at different stages of culture. After 3 days, outward currents were present, with an open probability increasing with depolarization, and the K+ channels showing a mean slope conductance of 82 pS in asymmetric and 168 pS in symmetric potassium solution. Tetraethylammonium (TEA) and quinidine blocked the channels. Cells at confluence showed similar channel activity, with conductances of 121 and 252 pS, respectively. We suggest that culture time and/or conditions may modify K+ channels or induce the expression of a new type of channels.

Action Potentials↗

PubMed1990

Modification of plasma membrane of differentiating preosseous chondrocytes: evidence for a degradative process in the mechanism of matrix vesicle formation.

Chondrocytes of the growth plate are differentiating cells. Their evolution leads to matrix vesicle formation and to cartilage mineralization. This is an in vitro study of the plasma membrane of chondrocytes at two differentiation stages. Differences in protein and glycoprotein components, increased membrane fluidity, and responsiveness to PTH indicate that hypertrophic ("ossifying") chondrocytes possess a plasma membrane widely different from that of resting chondrocytes. Their plasma membrane is particularly enriched in alkaline phosphatase (Mr 70K). Purified matrix vesicles contain the 70K form of alkaline phosphatase, but a 50K species is also detectable, a signal of degradative process. In fact, proteins and glycoproteins of matrix vesicles are less numerous than those of cell plasma membranes. It is suggested that, in vivo, matrix vesicle formation may be mediated by Ca2(+)-activated neutral proteases.

Alkaline Phosphatase↗

PubMedDate not supplied

[Solid-phase conglutinin tests for circulating IgA immunocomplexes].

In some renal, gastroenterological and dermatological diseases the pathogenic role of circulating immune complexes (CIC) of IgA type has been defined. The methods so far proposed to investigate CIC are for the most part able to identify IgGIC, while only few can do the same for IgAIC. The Authors discuss in details the technique and the sensibility and specificity of a new test for IgAIC based on the linkage with the conglutinin in solid phase and propose this test for the clinical study of IgA diseases.

Antigen-Antibody Complex↗