ELECTRICAL PROPERTIES OF OXYTOCIN NEURONS IN ORGANOTYPIC CULTURES
FROM POSTNATAL RAT HYPOTHALAMUS.
JOURDAIN, P., D. A. POULAIN, D. T. THEODOSIS and J. M. ISRAEL.
INSERM U. 378, Universit[umlaut]a Bordeaux 2, 146 rue L[umlaut]ao Saignat
F-33076 Bordeaux Cedex, France.
APStracts 3:0139N, 1996.
SUMMARY AND CONCLUSIONS
1. Intracellular recordings were performed on immunocytochemically-identified
oxytocin (OT) neurons (n = 101) maintained for 2 to 7 weeks in hypothalamic
organotypic cultures derived from 4 - 6 day-old rat neonates. The neurons
displayed a resting potential of -58.9 6.8 mV (n = 74), an input resistance
of 114 26.8 MW (n = 66) and a time constant of 9.6 1.4 ms (n = 57) (mean
SD). Voltage-current (V-I) relations, linear at resting potential, showed a
pronounced outward rectification when depolarized from hyperpolarized membrane
potentials. At these hyperpolarized potentials, depolarizing current pulses
induced a delayed action potential. 2. Action potentials had an amplitude of
73.4 9.7 mV and a duration of 1.9 0.2 ms. Each action potential was followed
by an afterhyperpolarization (AHP) of 7.9 2.0 mV in amplitude lasting 61.7
11.3 ms. The depolarizing phase of action potentials was both Na+ and Ca2+
dependent, whereas repolarization was due to a K+ conductance increase. 3.
When Ba2+ was substituted for Ca2+ in the medium, OT neurons displayed
prolonged sustained depolarizations. In the presence of TTX, these
depolarizations were triggered by depolarizing current pulses and arrested by
hyperpolarizing current pulses or by local application of Ca2+, Co2+, Cd2+. No
sustained depolarization was obtained when nifedipine was added to the medium.
These data suggest that OT cells in organotypic culture possess L-type Ca2+
channels. 5. All OT neurons generated spontaneous action potentials at resting
potential. Of 59 neurons, 29 showed a slow, irregular firing pattern (2.5
spikes/s), 24 generated a fast continuous firing pattern (³2.5 spikes/s) and 6
cells displayed a bursting pattern of activity, consisting of alternating
periods of spike discharge and quiescence. None of the bursting cells
exhibited regenerative endogenous potentials (plateau potentials). On the
contrary, in 4 of these cells, the bursting activity was clearly due to
patterned synaptic activity. 6. The cultured OT cells responded to exogenous
GABA and muscimol with a hyperpolarization and an increase in membrane
conductance. These effects were still observed in the presence of
tetrodotoxin, indicating that they were due to direct activation of GABA
receptors in the cells. The GABA-induced response was mediated by GABAA
receptors since it was blocked by bicuculline, but not by GABAB receptors,
since baclofen and hydroxysaclofen had no effect on membrane potential and
input resistance. 7. OT neurons responded to exogenous glutamate, quisqualate
and kainate with a depolarization concomitant with an increase in membrane
conductance. NMDA depolarized the cells in Mg2+-free medium. These effects
were observed in the presence of TTX, suggesting that OT cells expressed
ionotropic glutamate receptors. Trans-1S,3R-ACPD and MCPG had no effect on OT
cells, thus excluding the presence of metabotropic glutamate receptors. 8.
Taken together, our observations demonstrate that hypothalamic slice cultures
from 4 - 6 day-old rat neonates contain well differentiated OT neurons which
display electrical properties similar to those shown by adult neurons in
vitro. Such cultures provide a reliable model to investigate membrane
properties of adult OT neurons and a useful means to study the long-term
modulation of their electrical behaviour by various agents known to affect OT
cells in vivo.
Received 16 April 1996; accepted in final form 20 June 1996.
APS Manuscript Number J314-6.
Article publication pending J. Neurophysiol.
ISSN 1080-4757 Copyright 1996 The American Physiological Society.
Published in APStracts on 4 July 96