Abstract
The voltage-dependent potassium channel subunit Kv2.1 is widely expressed throughout the mammalian CNS and is clustered primarily on the somata and proximal dendrites, but not axons, of both principal neurones and inhibitory interneurones of the cortex and hippocampus. This expression pattern suggests that Kv2.1-containing channels may play a role in the regulation of pyramidal neurone excitability. To test this hypothesis and to determine the functional role of Kv2. 1-containing channels, cultured hippocampal slices were incubated with antisense oligonucleotides directed against Kv2.1 mRNA. Western blot analysis demonstrated that Kv2.1 protein content of cultured slices decreased > 90 % following 2 weeks of treatment with antisense oligonucleotides, when compared with either control missense-treated or untreated cultures. Similarly, Kv2.1 immunostaining was selectively decreased in antisense-treated cultures. Sustained outward potassium currents, recorded in both whole-cell and outside-out patch configurations, demonstrated a selective reduction of amplitude only in antisense-treated CA1 pyramidal neurones. Under current-clamp conditions, action potential durations were identical in antisense-treated, control missense-treated and untreated slices when initiated by low frequency stimulation (0.2 Hz). In contrast, spike repolarization was progressively prolonged during higher frequencies of stimulation (1 Hz) only in cells from antisense-treated slices. Similarly, action potentials recorded during electrographic interictal activity in the 'high [K+]o' model of epilepsy demonstrated pronounced broadening of their late phase only in cells from antisense-treated slices. Consistent with the frequency-dependent spike broadening, calcium imaging experiments from single CA1 pyramidal neurones revealed that high frequency Schaffer collateral stimulation resulted in a prolonged elevation of dendritic [Ca2+]i transients only in antisense-treated neurones. These studies demonstrate that channels containing Kv2.1 play a role in regulating pyramidal neurone somato-dendritic excitability primarily during episodes of high frequency synaptic transmission.
MeSH Terms
Action Potentials/physiology
Animals
Delayed Rectifier Potassium Channels
Dendrites/metabolism,physiology
Electric Conductivity
Electric Stimulation
Hippocampus/metabolism,physiology
Immunohistochemistry
In Vitro Techniques
Oligonucleotides, Antisense/pharmacology
Patch-Clamp Techniques
Potassium Channels/genetics,metabolism,physiology
Potassium Channels, Voltage-Gated
Pyramidal Cells/drug effects,physiology
Rats
Rats, Sprague-Dawley
Shab Potassium Channels
Synapses/physiology
Chemicals
Delayed Rectifier Potassium Channels
Kcnb1 protein, rat
Oligonucleotides, Antisense
Potassium Channels
Potassium Channels, Voltage-Gated
Shab Potassium Channels
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Du J
Laboratory of Cellular Neurophysiology, NICHD/NIH, Room 5A72, 49 Convent Drive, Bethesda, MD 20892-4995, USA.
Haak L L
Phillips-Tansey E
Russell J T
McBain C J
References (29)
29 references, click to expand
-
K+ channel regulation of signal propagation in dendrites of hippocampal pyramidal neurons.
Nature. 1997 Jun 26;387(6636):869-75
PMID: 9202119
-
Identification and functional characterization of a K+ channel alpha-subunit with regulatory properties specific to brain.
J Neurosci. 1997 Jun 15;17(12):4652-61
PMID: 9169526
-
Phosphorylation of the Kv2.1 K+ channel alters voltage-dependent activation.
Mol Pharmacol. 1997 Nov;52(5):821-8
PMID: 9351973
-
The K+ channel, Kv2.1, is apposed to astrocytic processes and is associated with inhibitory postsynaptic membranes in hippocampal and cortical principal neurons and inhibitory interneurons.
Neuroscience. 1998 May;84(1):37-48
PMID: 9522360
-
Functional and molecular differences between voltage-gated K+ channels of fast-spiking interneurons and pyramidal neurons of rat hippocampus.
J Neurosci. 1998 Oct 15;18(20):8111-25
PMID: 9763458
-
Memory and long-term potentiation (LTP) dissociated: normal spatial memory despite CA1 LTP elimination with Kv1.4 antisense.
Proc Natl Acad Sci U S A. 1998 Dec 8;95(25):15037-42
PMID: 9844011
-
Identification of the Kv2.1 K+ channel as a major component of the delayed rectifier K+ current in rat hippocampal neurons.
J Neurosci. 1999 Mar 1;19(5):1728-35
PMID: 10024359
-
A small domain in the N terminus of the regulatory alpha-subunit Kv2. 3 modulates Kv2.1 potassium channel gating.
J Neurosci. 1999 Aug 15;19(16):6865-73
PMID: 10436044
-
Arachidonic acid reciprocally alters the availability of transient and sustained dendritic K(+) channels in hippocampal CA1 pyramidal neurons.
J Neurosci. 1999 Oct 1;19(19):8163-71
PMID: 10493718
-
Outward currents of single hippocampal cells obtained from the adult guinea-pig.
J Physiol. 1987 Dec;393:331-53
PMID: 2451740
-
Rat hippocampal slices 'in vitro' display spontaneous epileptiform activity following long-term organotypic culture.
J Neurosci Methods. 1989 Feb;27(1):35-49
PMID: 2563782
-
A novel potassium channel with delayed rectifier properties isolated from rat brain by expression cloning.
Nature. 1989 Aug 24;340(6235):642-5
PMID: 2770868
-
A simple method for organotypic cultures of nervous tissue.
J Neurosci Methods. 1991 Apr;37(2):173-82
PMID: 1715499
-
Immunological identification and characterization of a delayed rectifier K+ channel polypeptide in rat brain.
Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10764-8
PMID: 1961744
-
Contrasting immunohistochemical localizations in rat brain of two novel K+ channels of the Shab subfamily.
J Neurosci. 1993 Apr;13(4):1569-76
PMID: 8463836
-
Properties of Kv2.1 K+ channels expressed in transfected mammalian cells.
J Biol Chem. 1994 Sep 16;269(37):23204-11
PMID: 8083226
-
Differential spatiotemporal expression of K+ channel polypeptides in rat hippocampal neurons developing in situ and in vitro.
J Neurosci. 1995 May;15(5 Pt 2):3840-51
PMID: 7751950
-
Elimination of potassium channel expression by antisense oligonucleotides in a pituitary cell line.
Proc Natl Acad Sci U S A. 1995 Jun 20;92(13):5955-9
PMID: 7597060
-
An inhibitor of the Kv2.1 potassium channel isolated from the venom of a Chilean tarantula.
Neuron. 1995 Oct;15(4):941-9
PMID: 7576642
-
Developmental expression and functional characterization of the potassium-channel subunit Kv3.1b in parvalbumin-containing interneurons of the rat hippocampus.
J Neurosci. 1996 Jan 15;16(2):506-18
PMID: 8551335
-
Voltage-gated potassium currents in stratum oriens-alveus inhibitory neurones of the rat CA1 hippocampus.
J Physiol. 1995 Nov 1;488 ( Pt 3):647-60
PMID: 8576855
-
Active properties of neuronal dendrites.
Annu Rev Neurosci. 1996;19:165-86
PMID: 8833440
-
Two temporally overlapping "delayed-rectifiers" determine the voltage-dependent potassium current phenotype in cultured hippocampal interneurons.
J Neurophysiol. 1996 Sep;76(3):1477-90
PMID: 8890268
-
Manipulation of the delayed rectifier Kv1.5 potassium channel in glial cells by antisense oligodeoxynucleotides.
Glia. 1996 Nov;18(3):177-84
PMID: 8915650
-
Identification of a cytoplasmic domain important in the polarized expression and clustering of the Kv2.1 K+ channel.
J Cell Biol. 1996 Dec;135(6 Pt 1):1619-32
PMID: 8978827
-
Action potential initiation and backpropagation in neurons of the mammalian CNS.
Trends Neurosci. 1997 Mar;20(3):125-31
PMID: 9061867
-
Modes of regulation of shab K+ channel activity by the Kv8.1 subunit.
J Biol Chem. 1997 Mar 28;272(13):8774-80
PMID: 9079713
-
Reversible antisense inhibition of Shaker-like Kv1.1 potassium channel expression impairs associative memory in mouse and rat.
Proc Natl Acad Sci U S A. 1997 Apr 29;94(9):4430-4
PMID: 9114006
-
Organotypic slice cultures: a technique has come of age.
Trends Neurosci. 1997 Oct;20(10):471-7
PMID: 9347615