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J L Brigant

Publications and source records attributed to J L Brigant.

8 recordsLinked to original sources
PubMed1988

Electrophysiology and ultrastructure of mouse hypothalamic neurons in culture: a correlative analysis during development.

The development of the electrical activity of hypothalamic neurons in dissociated cell cultures obtained from 14 day old mice foetuses was studied using patch extracellular and intracellular recording techniques. Electrophysiological data were compared with morphological observations obtained by electron microscopy. During patch recording, excitability of the cells was tested by the application of a 40 mM KCl solution. Tetrodotoxin (TTX, 10(-6) M in the delivery pipette) and Co2+ (10(-2) M in the delivery pipette) were applied to the recorded cell by pressure in order to study the involvement of sodium and calcium currents in the electrical activity during the in vitro development. From the first day of incubation, TTX and Co2+ were able to block reversibly the spontaneous electrical activity. However, only TTX application inhibited action potentials which suggests that calcium currents could be poorly involved in the action potential generation at the beginning of neuronal differentiation. Three different phases were found in the electrophysiological development of hypothalamic neurons in culture. The first phase (between the 1st and the 5th day of incubation) was characterized by an increase in the ratio of the spontaneously active cells (15% at day 1 and 90% at day 5). This increase paralleled the increase of the ratio of excitable cells. During this period no post-synaptic activity was detected. Morphologically, at 36 h, no synaptic contact was observed and growth cones were found to be very primitive. The second phase, between the 6th and the 9th day of culture, was characterized by a decrease in the ratio of spontaneously active cells and by the appearance, in a few cases, of a postsynaptic potential activity. During this phase the majority of the silent cells were excitable. At this stage neurons formed well differentiated neurites and growth cones. Synaptogenesis had already started and several stages of synapse formation could be seen. The third phase of the development, from 10 days of incubation, was characterized by an increase in post synaptic potential activity. During this period, numerous mature synapses could be observed although most of the synaptic contacts were located on neurites. In addition, some synapses were apposed onto degenerated structures. In conclusion, hypothalamic neurons in culture appear to differentiate in 3 steps: a primitive stage during which spontaneous electrical activity and excitability develop without any synaptic contact; a 2nd stage during which synaptic contacts develop, followed by a third stage of synapse maturation where mature synapses are formed whereas transient synapses degenerate.

Action Potentials↗
PubMed1987

[Development of electrical activity of hypothalamic cells in culture].

The development of the electrical activity of hypothalamic cells was studied using intracellular and patch clamp recording technics on cultured hypothalamic neurones from 14 days mouse embryos. After 24 h of incubation, 15% of recorded cells were spontaneously active. During the first five days of culture this ratio increased exponentially to reach 95% at day 5. Between the 5th and the 9th day the spontaneous activity progressively decreased although the majority of cells remained excitable. Spontaneous activity reappeared after the 9th day and was underlain by a synaptic potential activity. During the first five days of culture, only a TTX sensitive inward current was observed in all cell tested. A calcium inward current appeared after the first week of incubation. It was recorded on 40% of cells at day 11th and in 80% of cells tested after a month of incubation. Our results show that electrical activity of cultured hypothalamic neurones develop in three stages. A first stage characterized by a spontaneous electrical activity without post synaptic potential, an intermediate stage during which a Calcium inward current appeared corresponding to synaptic contact development and a third stage during which post synaptic potential activity was observed, corresponding to synaptic contact maturation.

Animals↗
PubMed1983

Botulinum A toxin and tetanus toxin do not affect presynaptic membrane currents in mammalian motor nerve endings.

The hypothesis according to which BoTx and TeTx block neuromuscular transmission by impairing Ca2+ influx was tested by recording presynaptic membrane currents at motor endplates of the mouse by means of external electrodes. The use of K-channel blockers allowed us to observe inward Ca current which was unaffected by bath application of BoTx or TeTx at doses which block neuromuscular transmission.

Animals↗
PubMed1982

Presynaptic currents in mouse motor endings.

1. We used external electrodes placed under precise visual control on motor endings of the mouse to record electrical activity promoted by nerve stimulation.2. Three types of wave form have been observed in relation to well-defined electrode emplacements: (i) at the transition between myelinated and non-myelinated parts of the axon, the wave form consists of two negative deflexions preceded by a small positivity (preterminal response), (ii) at the main part of the terminal branches, we obtained a two component positive wave form (terminal response) and (iii) electrode positions in a narrow area between the former and the latter yielded triphasic (positive-negative-positive) wave forms (intermediate responses).3. Since these responses could not be readily interpreted in terms of classical description of membrane currents associated with propagating action potentials, we used specific channel blocking agents to identify wave form components.4. Bath application of tetraethylammonium or aminopyridines, or, better, a combination of both, suppressed delayed positive deflexions of terminal and intermediate responses and the late negative component of preterminal responses. Local inophoretic drug application showed that K channels are present only at the terminal part of the endings. K(+) outflux promotes a local circuit whose sink is located at the preterminal part where it generates the late negative deflexion of the preterminal response.5. Local application of tetrodotoxin suppressed the first negative component of preterminal responses but failed to affect electrical activity at the terminal part of the endings. This indicates that Na channels, and, therefore, action potential generation, are restricted to the preterminal part.6. Suppression of K conductance revealed a slow inward current at the terminal part of the endings which could be identified as a Ca current. Ba(2+) and Sr(2+) could substitute for Ca(2+) as inward current carriers.7. Activation of spatially separated Na channels, on one side, and of K and Ca channels, on the other, generated ionic currents and separated local circuit currents which flow between preterminal and terminal parts (and vice versa). Thus, the signals recorded at each point of motor endings correspond to the sum of ionic and passive currents entering (or leaving) the membrane at that point.8. The present results represent a further example of heterogeneity of axonal membrane.

Action Potentials↗
PubMed1982

Electrical activity at motor nerve terminals of the mouse.

1. Extracellular recording of the electrical activity of mammalian end-plate made it possible to distinguish three different parts on the presynaptic terminal. Bath or ionophoretic application of ionic channel blockers induced specific alterations of electric signals which revealed the localization of classical ionic channels. 2. Sodium channels are restricted to a small area, sharply located after the end of the last myelinated segment and potassium channels are distributed over the rest of the terminal branches. 3. The first direct evidence of the presence of calcium channels at the terminal part of the motor endings was obtained, when potassium channel activity was suppressed. They occupy the same region as potassium channels. 4. Finally, the differential distribution of ionic channels over the terminal and the time-course of calcium current are discussed in relation to transmitter release.

Animals↗
PubMed1981

Advantages of the triangularis sterni muscle of the mouse for investigations of synaptic phenomena.

The triangularis sterni muscle (TS) of the mouse is a thin trapezoidal sheet of fibres in which individual neuromuscular junctions are easily observed with Nomarski optics. Thus, microelectrodes are readily positioned to accurately record various synaptic phenomena. For example, miniature end-plate currents were easily recorded with a focally positioned extracellular electrode and the end-plate sensitivity to acetylcholine averaged 2062 mV/nC. In addition, the intercostal nerves segmentally innervate the TS. Electrophysiologic and histologic analysis showed that each nerve innervates a sharply defined territory of the muscle surface. These preliminary observations suggest that the TS may be ideal for studies of synaptic function and the processes underlying synapse stabilization in the mammal.

Acetylcholine↗