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The Journal of Neurophysiology Vol. 83 No. 5 May 2000, pp. 2554-2561
Copyright ©2000 by the American Physiological Society
Department of Pharmacology, University College London, London WC1E 6BT, United Kingdom
Shah, M. and
D. G. Haylett.
Ca2+ Channels Involved in the Generation of the Slow
Afterhyperpolarization in Cultured Rat Hippocampal Pyramidal
Neurons. J. Neurophysiol. 83: 2554-2561, 2000. The advantages of using isolated cells have led us to develop
short-term cultures of hippocampal pyramidal cells, which retain many
of the properties of cells in acute preparations and in particular the
ability to generate afterhyperpolarizations after a train of action
potentials. Using perforated-patch recordings, both medium and slow
afterhyperpolarization currents (mIAHP and
sIAHP, respectively) could be obtained from
pyramidal cells that were cultured for 8-15 days. The
sIAHP demonstrated the kinetics and pharmacologic characteristics reported for pyramidal cells in slices.
In addition to confirming the insensitivity to 100 nM apamin and 1 mM
TEA, we have shown that the sIAHP is also
insensitive to 100 nM charybdotoxin but is inhibited by 100 µM
D-tubocurarine. Concentrations of nifedipine (10 µM) and
nimodipine (3 µM) that maximally inhibit L-type calcium channels
reduced the sIAHP by 30 and 50%,
respectively. However, higher concentrations of nimodipine (10 µM)
abolished the sIAHP, which can be partially
explained by an effect on action potentials. Both nifedipine and
nimodipine at maximal concentrations were found to reduce the HVA
calcium current in freshly dissociated neurons to the same extent. The N-type calcium channel inhibitor, 
-conotoxin GVIA (100 nM),
irreversibly inhibited the sIAHP by 37%.
Together, -conotoxin (100 nM) and nifedipine (10 µM) inhibited the
sIAHP by 70%. 10 µM ryanodine also
reduced the sIAHP by 30%, suggesting a role
for calcium-induced calcium release. It is concluded that activation of
the sIAHP in cultured hippocampal pyramidal
cells is mediated by a rise in intracellular calcium involving multiple
pathways and not just entry via L-type calcium channels.
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