Localization of SK2 channels relative to excitatory synaptic sites in the mouse developing Purkinje cells

Carmen Ballesteros-Merino, José Martínez-Hernández, Carolina Aguado, Masahiko Watanabe, John Adelman, Rafael Luján

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Small-conductance, Ca2+-activated K+ (SK) channels regulate neuronal excitability in a variety of ways. To understand their roles in different neuronal subtypes it is important to determine their precise subcellular distribution. Here, we used biochemical, light microscopy immunohistochemical and immunoelectron microscopy techniques, combined with quantitative approaches, to reveal the expression and subcellular localization patterns of SK2 in the developing cerebellum. Using western blots, the SK2 protein showed a progressive increase during postnatal development. At the light microscopic level, SK2 immunoreactivity was very prominent in the developing Purkinje cells (PC), particularly in the molecular layer (ML). Electron microscopy revealed that throughout development SK2 was mostly detected at the extrasynaptic and perisynaptic plasma membrane of dendritic shafts and dendritic spines of PCs. However, there was some localization at axon terminals as well. Quantitative analyses and 3D reconstructions further revealed a progressive developmental change of SK2 on the surface of PCs from dendritic shafts to dendritic spines. Together, these results indicate that SK2 channels undergo dynamic spatial regulation during cerebellar development, and this process is associated with the formation and maturation of excitatory synaptic contacts to PCs.

Original languageEnglish (US)
Article number154
JournalFrontiers in Neuroanatomy
Volume8
Issue numberDEC
DOIs
StatePublished - Dec 15 2014

Fingerprint

Dendritic Spines
Purkinje Cells
Light
Calcium-Activated Potassium Channels
Immunoelectron Microscopy
Presynaptic Terminals
Cerebellum
Microscopy
Electron Microscopy
Western Blotting
Cell Membrane
Proteins

Keywords

  • Cerebellar development
  • Electron microscopy
  • Immunohistochemistry
  • Purkinje cells
  • SK2 channels

ASJC Scopus subject areas

  • Anatomy
  • Neuroscience (miscellaneous)
  • Cellular and Molecular Neuroscience

Cite this

Ballesteros-Merino, C., Martínez-Hernández, J., Aguado, C., Watanabe, M., Adelman, J., & Luján, R. (2014). Localization of SK2 channels relative to excitatory synaptic sites in the mouse developing Purkinje cells. Frontiers in Neuroanatomy, 8(DEC), [154]. https://doi.org/10.3389/fnana.2014.00154

Localization of SK2 channels relative to excitatory synaptic sites in the mouse developing Purkinje cells. / Ballesteros-Merino, Carmen; Martínez-Hernández, José; Aguado, Carolina; Watanabe, Masahiko; Adelman, John; Luján, Rafael.

In: Frontiers in Neuroanatomy, Vol. 8, No. DEC, 154, 15.12.2014.

Research output: Contribution to journalArticle

Ballesteros-Merino, C, Martínez-Hernández, J, Aguado, C, Watanabe, M, Adelman, J & Luján, R 2014, 'Localization of SK2 channels relative to excitatory synaptic sites in the mouse developing Purkinje cells', Frontiers in Neuroanatomy, vol. 8, no. DEC, 154. https://doi.org/10.3389/fnana.2014.00154
Ballesteros-Merino C, Martínez-Hernández J, Aguado C, Watanabe M, Adelman J, Luján R. Localization of SK2 channels relative to excitatory synaptic sites in the mouse developing Purkinje cells. Frontiers in Neuroanatomy. 2014 Dec 15;8(DEC). 154. https://doi.org/10.3389/fnana.2014.00154
Ballesteros-Merino, Carmen ; Martínez-Hernández, José ; Aguado, Carolina ; Watanabe, Masahiko ; Adelman, John ; Luján, Rafael. / Localization of SK2 channels relative to excitatory synaptic sites in the mouse developing Purkinje cells. In: Frontiers in Neuroanatomy. 2014 ; Vol. 8, No. DEC.
@article{14134423ccee4c29a161270196fcb1aa,
title = "Localization of SK2 channels relative to excitatory synaptic sites in the mouse developing Purkinje cells",
abstract = "Small-conductance, Ca2+-activated K+ (SK) channels regulate neuronal excitability in a variety of ways. To understand their roles in different neuronal subtypes it is important to determine their precise subcellular distribution. Here, we used biochemical, light microscopy immunohistochemical and immunoelectron microscopy techniques, combined with quantitative approaches, to reveal the expression and subcellular localization patterns of SK2 in the developing cerebellum. Using western blots, the SK2 protein showed a progressive increase during postnatal development. At the light microscopic level, SK2 immunoreactivity was very prominent in the developing Purkinje cells (PC), particularly in the molecular layer (ML). Electron microscopy revealed that throughout development SK2 was mostly detected at the extrasynaptic and perisynaptic plasma membrane of dendritic shafts and dendritic spines of PCs. However, there was some localization at axon terminals as well. Quantitative analyses and 3D reconstructions further revealed a progressive developmental change of SK2 on the surface of PCs from dendritic shafts to dendritic spines. Together, these results indicate that SK2 channels undergo dynamic spatial regulation during cerebellar development, and this process is associated with the formation and maturation of excitatory synaptic contacts to PCs.",
keywords = "Cerebellar development, Electron microscopy, Immunohistochemistry, Purkinje cells, SK2 channels",
author = "Carmen Ballesteros-Merino and Jos{\'e} Mart{\'i}nez-Hern{\'a}ndez and Carolina Aguado and Masahiko Watanabe and John Adelman and Rafael Luj{\'a}n",
year = "2014",
month = "12",
day = "15",
doi = "10.3389/fnana.2014.00154",
language = "English (US)",
volume = "8",
journal = "Frontiers in Neuroanatomy",
issn = "1662-5129",
publisher = "Frontiers Research Foundation",
number = "DEC",

}

TY - JOUR

T1 - Localization of SK2 channels relative to excitatory synaptic sites in the mouse developing Purkinje cells

AU - Ballesteros-Merino, Carmen

AU - Martínez-Hernández, José

AU - Aguado, Carolina

AU - Watanabe, Masahiko

AU - Adelman, John

AU - Luján, Rafael

PY - 2014/12/15

Y1 - 2014/12/15

N2 - Small-conductance, Ca2+-activated K+ (SK) channels regulate neuronal excitability in a variety of ways. To understand their roles in different neuronal subtypes it is important to determine their precise subcellular distribution. Here, we used biochemical, light microscopy immunohistochemical and immunoelectron microscopy techniques, combined with quantitative approaches, to reveal the expression and subcellular localization patterns of SK2 in the developing cerebellum. Using western blots, the SK2 protein showed a progressive increase during postnatal development. At the light microscopic level, SK2 immunoreactivity was very prominent in the developing Purkinje cells (PC), particularly in the molecular layer (ML). Electron microscopy revealed that throughout development SK2 was mostly detected at the extrasynaptic and perisynaptic plasma membrane of dendritic shafts and dendritic spines of PCs. However, there was some localization at axon terminals as well. Quantitative analyses and 3D reconstructions further revealed a progressive developmental change of SK2 on the surface of PCs from dendritic shafts to dendritic spines. Together, these results indicate that SK2 channels undergo dynamic spatial regulation during cerebellar development, and this process is associated with the formation and maturation of excitatory synaptic contacts to PCs.

AB - Small-conductance, Ca2+-activated K+ (SK) channels regulate neuronal excitability in a variety of ways. To understand their roles in different neuronal subtypes it is important to determine their precise subcellular distribution. Here, we used biochemical, light microscopy immunohistochemical and immunoelectron microscopy techniques, combined with quantitative approaches, to reveal the expression and subcellular localization patterns of SK2 in the developing cerebellum. Using western blots, the SK2 protein showed a progressive increase during postnatal development. At the light microscopic level, SK2 immunoreactivity was very prominent in the developing Purkinje cells (PC), particularly in the molecular layer (ML). Electron microscopy revealed that throughout development SK2 was mostly detected at the extrasynaptic and perisynaptic plasma membrane of dendritic shafts and dendritic spines of PCs. However, there was some localization at axon terminals as well. Quantitative analyses and 3D reconstructions further revealed a progressive developmental change of SK2 on the surface of PCs from dendritic shafts to dendritic spines. Together, these results indicate that SK2 channels undergo dynamic spatial regulation during cerebellar development, and this process is associated with the formation and maturation of excitatory synaptic contacts to PCs.

KW - Cerebellar development

KW - Electron microscopy

KW - Immunohistochemistry

KW - Purkinje cells

KW - SK2 channels

UR - http://www.scopus.com/inward/record.url?scp=84918807261&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84918807261&partnerID=8YFLogxK

U2 - 10.3389/fnana.2014.00154

DO - 10.3389/fnana.2014.00154

M3 - Article

VL - 8

JO - Frontiers in Neuroanatomy

JF - Frontiers in Neuroanatomy

SN - 1662-5129

IS - DEC

M1 - 154

ER -