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1 Bogomoletz Institute of Physiology, 4 Bogomoletz Street, Kiev, 01024, Ukraine
2 Department of Physiology, University of Cambridge, CB2 3EG, UK
Increases in Ca2+ concentration in the nucleus of neurones modulate gene transcription and may be involved in activity-dependent long-term plasticity, apoptosis, and neurotoxicity. Little is currently known about the regulation of Ca2+ in the nuclei of neurones. Investigation of neuronal nuclei is hampered by the cellular heterogeneity of the brain where neurones comprise no more than 10% of the cells. The situation is further complicated by large differences in properties of different neurones. Here we report a method for isolating nuclei from identified central neurones. We employed this technique to study nuclei from rat cerebellar Purkinje and granule neurones. Patch-clamp recording from the nuclear membrane of Purkinje neurones revealed numerous large-conductance channels selective for monovalent cations. The nuclear membrane of Purkinje neurones also contained multiple InsP3- activated ion channels localized exclusively in the inner nuclear membrane with their receptor loci facing the nucleoplasm. In contrast, the nuclear membrane of granule neurones contained only a small number of mainly anion channels. Nuclear InsP3 receptors (InsP3Rs) were activated by InsP3 with EC50 = 0.67 µM and a Hill coefficient of 2.5. Ca2+ exhibited a biphasic effect on the receptors elevating its activity at low concentrations and inhibiting it at micromolar concentrations. InsP3 in saturating concentrations did not prevent the inhibitory effect of Ca2+, but strongly increased InsP3R activity at resting Ca2+ concentrations. These data are the first evidence for the presence of intranuclear sources of Ca2+ in neurones. Ca2+ release from the nuclear envelope may amplify Ca2+ transients penetrating the nucleus from the cytoplasm or generate Ca2+ transients in the nucleus independently of the cytoplasm.
(Received 14 December 2004;
accepted after revision 10 March 2005;
first published online 17 March 2005)
Corresponding author S. M. Marchenko: Bogomoletz Institute of Physiology, 4 Bogomoletz Street, Kiev, 01024, Ukraine. Email: smm{at}serv.biph.kiev.ua
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