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J Physiol Volume 554, Number 3, 673-685, February 1, 2004 DOI: 10.1113/jphysiol.2003.056192
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Focal aggregation of voltage-gated, Kv2.1 subunit-containing, potassium channels at synaptic sites in rat spinal motoneurones

Elizabeth A. L. Muennich and R. E. W. Fyffe

Department of Anatomy and Physiology, Wright State University, Dayton, OH 45435, USA

Delayed rectifier K+ currents are involved in the control of {alpha}-motoneurone excitability, but the precise spatial distribution and organization of the membrane ion channels that contribute to these currents have not been defined. Voltage-activated Kv2.1 channels have properties commensurate with a contribution to delayed rectifier currents and are expressed in neurones throughout the mammalian central nervous system. A specific antibody against Kv2.1 channel subunits was used to determine the surface distribution and clustering of Kv2.1 subunit-containing channels in the cell membrane of {alpha}-motoneurones and other spinal cord neurones. In {alpha}-motoneurones, Kv2.1 immunoreactivity (-IR) was abundant in the surface membrane of the soma and large proximal dendrites, and was present also in smaller diameter distal dendrites. Plasma membrane-associated Kv2.1-IR in {alpha}-motoneurones was distributed in a mosaic of small irregularly shaped, and large disc-like, clusters. However, only small to medium clusters of Kv2.1-IR were observed in spinal interneurones and projection neurones, and some interneurones, including Renshaw cells, lacked demonstrable Kv2.1-IR. In {alpha}-motoneurones, dual immunostaining procedures revealed that the prominent disc-like domains of Kv2.1-IR are invariably apposed to presynaptic cholinergic C-terminals. Further, Kv2.1-IR colocalizes with immunoreactivity against postsynaptic muscarinic (m2) receptors at these locations. Ultrastructural examination confirmed the postsynaptic localization of Kv2.1-IR at C-terminal synapses, and revealed clusters of Kv2.1-IR at a majority of S-type, presumed excitatory, synapses. Kv2.1-IR in {alpha}-motoneurones is not directly associated with presumed inhibitory (F-type) synapses, nor is it present in presynaptic structures apposed to the motoneurone. Occasionally, small patches of extrasynaptic Kv2.1-IR labelling were observed in surface membrane apposed by glial processes. Voltage-gated potassium channels responsible for the delayed rectifier current, including Kv2.1, are usually assigned roles in the repolarization of the action potential. However, the strategic localization of Kv2.1 subunit-containing channels at specific postsynaptic sites suggests that this family of voltage-activated K+ channels may have additional roles and/or regulatory components.

(Received 30 September 2003; accepted after revision 5 November 2003; first published online 7 November 2003)
Corresponding author R. E. W. Fyffe: Department of Anatomy and Physiology, Wright State University, Dayton, OH 45435, USA.  Email: robert.fyffe{at}wright.edu




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