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N Chuhma

Publications and source records attributed to N Chuhma.

6 recordsLinked to original sources

Differential development of Ca2+ dynamics in presynaptic terminal and postsynaptic neuron of the rat auditory synapse.

Postnatal development of Ca2+ influx and Ca2+ clearance capacity were investigated in the synapse of medial nucleus of the trapezoid body (MNTB) of rat with fura-2 fluorimetry. In contrast to the presynaptic terminal, Ca2+ dynamics does not basically change in the postsynaptic principal neuron developmentally. This differential development of Ca2+ dynamics between pre- and postsynaptic neurons might be crucial for the organized formation and functional maturation of this synapse.

Animals↗

Synchronisation of neurotransmitter release during postnatal development in a calyceal presynaptic terminal of rat.

Mechanisms contributing to the synchronisation of transmitter release during development were studied in synapses of the medial nucleus of the trapezoid body (MNTB) using patch recording and Ca2+ imaging techniques in a rat brainstem slice preparation. Excitatory postsynaptic currents (EPSCs) were generated in an all-or-none manner at immature synapses (postnatal days earlier than P6). Many delayed miniature EPSC (mEPSC)-like currents followed EPSCs at immature synapses, while observations of delayed mEPSC-like currents were rare at mature synapses (later than P9). At immature synapses bath application of either omega-conotoxin GVIA or omega-agatoxin-IVA reduced EPSCs (both to 40% of control), and Ca2+ currents in the presynaptic terminal (both to 70% of control). The frequency of delayed mEPSC-like currents was reduced by omega-conotoxin GVIA, but not by omega-agatoxin IVA. At immature synapses delayed mEPSC-like currents were rare after incubation of the slice with extrinsic Ca2+ buffers (EGTA AM). At mature synapses many mEPSC-like currents followed evoked EPSCs after partial block of Ca2+ channels by bath application of a low concentration of Cd2+ (3 microM) or omega-agatoxin IVA (50 nM) but not by low [Ca2+]o (0.5-1 mM). Ca2+ transients induced by action potentials in presynaptic terminals were monitored by adding a high concentration of fura-2 (200 microM) to the pipette. Their decay time course was slower at immature presynaptic terminals than at mature terminals. Both the Ca2+ extrusion rate and the endogenous Ca2+ binding capacity were estimated to be smaller at immature terminals than at mature terminals. These results suggest that the maturation of synaptic transmission in MNTB progresses with the capacity for Ca2+ clearance from the presynaptic terminal. The possible importance of developmental increases in both Ca2+ clearance capacity and Ca2+ currents is discussed in relation to the synchronisation of transmitter release.

Animals↗

Postnatal development of phase-locked high-fidelity synaptic transmission in the medial nucleus of the trapezoid body of the rat.

Synaptic transmission in the medial nucleus of the trapezoid body of rats was analyzed in postnatal days 4-13 (P4-P13) by applying the whole-cell patch-recording technique to brain slices. In P4-P6 animals, evoked EPSCs fluctuated extensively in amplitude and occurred in marked asynchrony, followed by spontaneous EPSCs. With development of animals, the evoked EPSCs increased in amplitude, and the rise time became faster. In addition, the synaptic transmission became phase-locked. The coefficient of variation (CV) of EPSC amplitude decreased with development (0.32 +/- 0.03 for P4-P5 and 0. 05 +/- 0.01 for P9-P11). The amplitude of miniature EPSCs did not change throughout the postnatal days investigated (-30.2 +/- 0.3 pA at -70 mV). The CV was dependent on extracellular Ca2+ concentration ([Ca2+]o) and was reduced with the increase of [Ca2+]o, and this [Ca2+]o dependence was shifted toward lower [Ca2+]o with development. Direct patch recording of the presynaptic terminals demonstrated an increase in Ca2+ currents during these postnatal days. The phase-locked high-fidelity transmission in this synapse is achieved with development likely through the increase of Ca2+ currents and Ca2+ sensitivity of transmitter release mechanisms in the presynaptic terminal.

Action Potentials↗

Omega-conotoxin-sensitive and -resistant transmitter release from the chick ciliary presynaptic terminal.

1. Synaptically evoked responses to stimulation of the oculomotor nerve were recorded from the ciliary nerve in chick embryos. The postsynaptic currents in response to presynaptic stimulation (EPSCs) were also recorded under whole-cell voltage clamp of the ciliary cell. 2. The ciliary nerve response was dependent on the extracellular Ca2+ concentration ([Ca2+]o). omega-Conotoxin GVIA (omega-CgTX, 100 nM) increased the [Ca2+]o necessary to evoke the half-maximal response by a factor of 1.7 without changing the slope of [Ca2+]o dependence. Dihydropyridine (DHP) derivatives, nifedipine or Bay K 8644, did not affect the [Ca2+]o sensitivity of ciliary nerve response. 3. The EPSC was usually preceded by the capacitive coupling response of the presynaptic action potential. In some records, the EPSCs were also preceded by the electrical coupling responses which were the mirror images of the presynaptic action potentials. The current-voltage relation of the EPSCs showed inward rectification. 4. The EPSC was potentiated by 4-aminopyridine (4-AP) as a result of prolongation of the falling phase of presynaptic action potential. In the presence of high [Ca2+]o and 4-AP, a small fraction of EPSC was resistant to omega-CgTX. 5. The resting potential of the presynaptic terminal was changed from -69 to -57 mV by increasing [K+]o from 1 to 10 mM. The same procedure decreased the omega-CgTX-resistant EPSC by 30%, whereas the omega-CgTX-untreated EPSC in low-Ca2+ saline was not affected by the change in [K+]o. 6. The nerve-evoked increase in intracellular Ca2+ was recorded from the presynaptic terminal (delta[Ca2+]pre). The delta[Ca2+]pre was larger in a solution containing 10 mM Ca2+ and 1 mM K+ after treating with omega-CgTX than in a solution containing 2 mM Ca2+ and 16 mM Mg2+ before treating with omega-CgTX. The EPSC was, in contrast, smaller in the 10 mM Ca(2+)-1 mM K+ solution after omega-CgTX treatment than in the 2 mM Ca(2+)-16 mM Mg2+ solution before omega-CgTX treatment. 7. Similarly, the EPSC was smaller in the 10 mM Ca(2+)-1 mM K+ solution containing 5 microM La3+ than in the 2 mM Ca(2+)-16 mM Mg2+ solution, whereas the delta [Ca2+]pre was larger in the 10 mM Ca(2+)-1 mM K+ solution containing 5 micrograms La3+ than in the 2 mM Ca(2+)-16 mM Mg2+ solution. 8. It is concluded that the omega-CgTX-sensitive Ca2+ conductance of the presynaptic terminal is the principal source of Ca2+ involved in transmitter release.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Preferential inhibition of omega-conotoxin-sensitive presynaptic Ca2+ channels by adenosine autoreceptors.

Adenosine is a potent modulator of transmitter release at a variety of synapses. The adenosine A1 receptor is assumed to reside in presynaptic terminals and to function as a negative autoreceptor. How adenosine reduces transmitter release is uncertain; it may reduce the calcium influx during nerve terminal depolarization by either activating K+ currents or inhibiting Ca2+ currents, although other mechanisms have been proposed. We have directly measured intracellular Ca2+ concentrations of giant pre-synaptic terminals in the chick ciliary ganglion. We report here that adenosine inhibited the nerve-evoked Ca2+ influx in the terminal by activating A1 receptors. Reduced Ca2+ influx was due largely to inhibition of omega-conotoxin GVIA-sensitive Ca2+ channels in the presynaptic terminal.

Adenosine↗

An improved method for perforated patch recordings using nystatin-fluorescein mixture.

This paper describes an improved method for solubilizing nystatin in an aqueous solution without the aid of organic solvents. The patch pipette was filled with a solution of nystatin and fluorescein sodium mixed in a molar ratio of 1:10. The success rate of perforated patch recordings was substantially improved, and the access resistance of 20-40 M omega was readily achieved.

Action Potentials↗