Light and electron microscopic analysis of KChIP and Kv4 localization in rat cerebellar granule cells

Brian W. Strassle, Milena Menegola, Kenneth J. Rhodes, James Trimmer

Research output: Contribution to journalArticle

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Abstract

Potassium channels are key determinants of neuronal excitability. We recently identified KChIPs as a family of calcium binding proteins that coassociate and colocalize with Kv4 family potassium channels in mammalian brain (An et al. [2000] Nature 403:553). Here, we used light microscopic immunohistochemistry and multilabel immunofluorescence labeling, together with transmission electron microscopic immunohistochemistry, to examine the subcellular distribution of KChIPs and Kv4 channels in adult rat cerebellum. Light microscopic immunohistochemistry was performed on 40-μm free-floating sections using a diaminobenzidine labeling procedure. Multilabel immunofluorescence staining was performed on free-floating sections and on 1-μm ultrathin cryosections. Electron microscopic immunohistochemistry was performed using an immunoperoxidase pre-embedding labeling procedure. By light microscopy, immunoperoxidase labeling showed that Kv4.2, Kv4.3, and KChIPs 1, 3, and 4 (but not KChIP2) were expressed at high levels in cerebellar granule cells (GCs). Kv4.2 and KChIP1 were highly expressed in GCs in rostral cerebellum, whereas Kv4.3 was more highly expressed in GCs in caudal cerebellum. Immunofluorescence labeling revealed that KChIP1 and Kv4.2 are concentrated in somata of cerebellar granule cells and in synaptic glomeruli that surround synaptophysin-positive mossy fiber axon terminals. Electron microscopic analysis revealed that KChIP1 and Kv4.2 immunoreactivity is concentrated along the plasma membrane of cerebellar granule cell somata and dendrites. In synaptic glomeruli, KChIP1 and Kv4.2 immunoreactivity is concentrated along the granule cell dendritic membrane, but is not concentrated at postsynaptic densities. Taken together, these data suggest that A-type potassium channels containing Kv4.2 and KChIP1, and perhaps also KChIP3 and 4, play a critical role in regulating postsynaptic excitability at the cerebellar mossy-fiber/ granule cell synapse.

Original languageEnglish (US)
Pages (from-to)144-155
Number of pages12
JournalJournal of Comparative Neurology
Volume484
Issue number2
DOIs
StatePublished - Apr 4 2005

Fingerprint

Electrons
Light
Shal Potassium Channels
Immunohistochemistry
Cerebellum
Fluorescent Antibody Technique
Carisoprodol
Cell Membrane
Post-Synaptic Density
Synaptophysin
Calcium-Binding Proteins
Potassium Channels
Presynaptic Terminals
Dendrites
Nerve Fibers
Synapses
Microscopy
Staining and Labeling
Brain

Keywords

  • A current
  • Cerebellum
  • Dendrite
  • Excitability
  • Shal
  • Synaptic plasticity

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Light and electron microscopic analysis of KChIP and Kv4 localization in rat cerebellar granule cells. / Strassle, Brian W.; Menegola, Milena; Rhodes, Kenneth J.; Trimmer, James.

In: Journal of Comparative Neurology, Vol. 484, No. 2, 04.04.2005, p. 144-155.

Research output: Contribution to journalArticle

Strassle, Brian W. ; Menegola, Milena ; Rhodes, Kenneth J. ; Trimmer, James. / Light and electron microscopic analysis of KChIP and Kv4 localization in rat cerebellar granule cells. In: Journal of Comparative Neurology. 2005 ; Vol. 484, No. 2. pp. 144-155.
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AB - Potassium channels are key determinants of neuronal excitability. We recently identified KChIPs as a family of calcium binding proteins that coassociate and colocalize with Kv4 family potassium channels in mammalian brain (An et al. [2000] Nature 403:553). Here, we used light microscopic immunohistochemistry and multilabel immunofluorescence labeling, together with transmission electron microscopic immunohistochemistry, to examine the subcellular distribution of KChIPs and Kv4 channels in adult rat cerebellum. Light microscopic immunohistochemistry was performed on 40-μm free-floating sections using a diaminobenzidine labeling procedure. Multilabel immunofluorescence staining was performed on free-floating sections and on 1-μm ultrathin cryosections. Electron microscopic immunohistochemistry was performed using an immunoperoxidase pre-embedding labeling procedure. By light microscopy, immunoperoxidase labeling showed that Kv4.2, Kv4.3, and KChIPs 1, 3, and 4 (but not KChIP2) were expressed at high levels in cerebellar granule cells (GCs). Kv4.2 and KChIP1 were highly expressed in GCs in rostral cerebellum, whereas Kv4.3 was more highly expressed in GCs in caudal cerebellum. Immunofluorescence labeling revealed that KChIP1 and Kv4.2 are concentrated in somata of cerebellar granule cells and in synaptic glomeruli that surround synaptophysin-positive mossy fiber axon terminals. Electron microscopic analysis revealed that KChIP1 and Kv4.2 immunoreactivity is concentrated along the plasma membrane of cerebellar granule cell somata and dendrites. In synaptic glomeruli, KChIP1 and Kv4.2 immunoreactivity is concentrated along the granule cell dendritic membrane, but is not concentrated at postsynaptic densities. Taken together, these data suggest that A-type potassium channels containing Kv4.2 and KChIP1, and perhaps also KChIP3 and 4, play a critical role in regulating postsynaptic excitability at the cerebellar mossy-fiber/ granule cell synapse.

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