Immunolocalization of the Ca2+-activated K+ channel Slo1 in axons and nerve terminals of mammalian brain and cultured neurons

Hiroaki Misonou, Milena Menegola, Lynn Buchwalder, Eunice W. Park, Andrea Meredith, Kenneth J. Rhodes, Richard W. Aldrich, James Trimmer

Research output: Contribution to journalArticle

93 Citations (Scopus)

Abstract

Ca2+-activated voltage-dependent K+ channels (Slo1, KCa1.1, Maxi-K, or BK channel) play a crucial role in controlling neuronal signaling by coupling channel activity to both membrane depolarization and intracellular Ca2+ signaling. In mammalian brain, immunolabeling experiments have shown staining for Slo1 channels predominantly localized to axons and presynaptic terminals of neurons. We have developed anti-Slo1 mouse monoclonal antibodies that have been extensively characterized for specificity of staining against recombinant Slo1 in heterologous cells, and native Slo1 in mammalian brain, and definitively by the lack of detectable immunoreactivity against brain samples from Slo1 knockout mice. Here we provide precise immunolocalization of Slo1 in rat brain with one of these monoclonal antibodies and show that Slo1 is accumulated in axons and synaptic terminal zones associated with glutamatergic synapses in hippocampus and GABAergic synapses in cerebellum. By using cultured hippocampal pyramidal neurons as a model system, we show that heterologously expressed Slo1 is initially targeted to the axonal surface membrane, and with further development in culture, become localized in presynaptic terminals. These studies provide new insights into the polarized localization of Slo1 channels in mammalian central neurons and provide further evidence for a key role in regulating neurotransmitter release in glutamatergic and GABAergic terminals.

Original languageEnglish (US)
Pages (from-to)289-302
Number of pages14
JournalJournal of Comparative Neurology
Volume496
Issue number3
DOIs
StatePublished - May 20 2006

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Calcium-Activated Potassium Channels
Presynaptic Terminals
Neurons
Large-Conductance Calcium-Activated Potassium Channels
Brain
Synapses
Monoclonal Antibodies
Staining and Labeling
Intracellular Membranes
Pyramidal Cells
Knockout Mice
Cerebellum
Neurotransmitter Agents
Hippocampus
Membranes

Keywords

  • BK channel
  • Ion channel
  • Localization
  • mAb
  • Protein trafficking

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Misonou, H., Menegola, M., Buchwalder, L., Park, E. W., Meredith, A., Rhodes, K. J., ... Trimmer, J. (2006). Immunolocalization of the Ca2+-activated K+ channel Slo1 in axons and nerve terminals of mammalian brain and cultured neurons. Journal of Comparative Neurology, 496(3), 289-302. https://doi.org/10.1002/cne.20931

Immunolocalization of the Ca2+-activated K+ channel Slo1 in axons and nerve terminals of mammalian brain and cultured neurons. / Misonou, Hiroaki; Menegola, Milena; Buchwalder, Lynn; Park, Eunice W.; Meredith, Andrea; Rhodes, Kenneth J.; Aldrich, Richard W.; Trimmer, James.

In: Journal of Comparative Neurology, Vol. 496, No. 3, 20.05.2006, p. 289-302.

Research output: Contribution to journalArticle

Misonou, H, Menegola, M, Buchwalder, L, Park, EW, Meredith, A, Rhodes, KJ, Aldrich, RW & Trimmer, J 2006, 'Immunolocalization of the Ca2+-activated K+ channel Slo1 in axons and nerve terminals of mammalian brain and cultured neurons', Journal of Comparative Neurology, vol. 496, no. 3, pp. 289-302. https://doi.org/10.1002/cne.20931
Misonou, Hiroaki ; Menegola, Milena ; Buchwalder, Lynn ; Park, Eunice W. ; Meredith, Andrea ; Rhodes, Kenneth J. ; Aldrich, Richard W. ; Trimmer, James. / Immunolocalization of the Ca2+-activated K+ channel Slo1 in axons and nerve terminals of mammalian brain and cultured neurons. In: Journal of Comparative Neurology. 2006 ; Vol. 496, No. 3. pp. 289-302.
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