Cellular neuroscienceDevelopmental localization of potassium chloride co-transporter 2 in granule cells of the early postnatal mouse cerebellum with special reference to the synapse formation
Section snippets
Animals
We examined mice from the C57Bl/6CrSlc mice of P0, P3, P5, 7, 10, 12, 14, 21 and 90 (as an adult). At each age, at least five mice (three for light microscopic analysis, two for electron microscopic analysis) were killed for immunohistochemistry.
Establishment of the KCC2 antibody
A cysteine residue was introduced at the amino terminal of the synthesized peptide, PVS SEG IKD FFS MKP EWE NLN, which are amino acid residues 1022–1042 of the mouse KCC2 (AF332064.1, GI:14193695). The peptide was conjugated with keyhole limpet
Immunohistochemical localization of KCC2 in the adult mouse cerebellar cortex
The specificity of the rabbit and guinea-pig KCC2 antibodies was examined by immunoblot analysis and immunohistochemistry. Each immunoblot showed a major band around the predicted molecular weight (140 kD), and the band was abolished by addition of the peptide for immunization into the primary antibody solution (Fig. 1A).
The immunohistochemical staining with rabbit KCC2-antibody showed that the immunolabeling was observed in the all three layers (Fig. 1B). In the molecular layer and Purkinje
Discussion
In the present study, we found that the KCC2 was first localized at the granule cell somata and dendrites, which formed synapses with synaptophysin-positive mossy fiber terminals at P5 as summarized in Fig. 6. The KCC2-positive granule cells increased in number during development, and all granule cells became positive by P21.
Acknowledgments
We are grateful to Hideki Nakamura at the Central Research Department of Hokkaido University School of Medicine for the technical assistance. We also thank to Dr. Hitoshi Komuro at Cleveland Clinic Foundation and Sachiko Yoshida at Toyohashi University of Technology for the valuable discussion.
References (45)
- et al.
GABA itself promotes the developmental switch of neuronal GABAergic responses from excitation to inhibition
Cell
(2001) - et al.
Molecular cloning and functional expression of the K-Cl cotransporter from rabbit, rat, and humanA new member of the cation-chloride cotransporter family
J Biol Chem
(1996) - et al.
Disruption of KCC2 reveals an essential role of K-Cl cotransport already in early synaptic inhibition
Neuron
(2001) - et al.
The differential expression patterns of messenger RNAs encoding K-Cl cotransporters (KCC1,2) and Na-K-2Cl cotransporter (NKCC1) in the rat nervous system
Neuroscience
(2001) - et al.
Getting excited about GABA and sex differences in the brain
Trends Neurosci
(2002) - et al.
An autoradiographic analysis of histogenesis in the mouse cerebellum
Exp Neurol
(1961) - et al.
Developmental changes in KCC1, KCC2 and NKCC1 mRNAs in the rat cerebellum
Brain Res Dev Brain Res
(2002) - et al.
Cation-chloride co-transporters in neuronal communication, development and trauma
Trends Neurosci
(2003) - et al.
Molecular characterization of a putative K-Cl cotransporter in rat brainA neuronal-specific isoform
J Biol Chem
(1996) - et al.
Trophic actions of GABA on neuronal development
Trends Neurosci
(2005)
GABAergic signaling in the developing cerebellum
Int Rev Neurobiol
Normal formation of the postsynaptic elements of GABAergic synapses in the reeler cerebellum
Brain Res Dev Brain Res
Extrasynaptic localization of GABA in the developing mouse cerebellum
Neurosci Res
Morphological development and maturation of the GABAergic synapses in the mouse cerebellar granular layer
Brain Res Dev Brain Res
Transient expression of GABA(A) receptor alpha2 and alpha3 subunits in differentiating cerebellar neurons
Brain Res Dev Brain Res
Developmental expression of GABA transporter-1 and 3 during formation of the GABAergic synapses in the mouse cerebellar cortex
Brain Res Dev Brain Res
Developmental changes in KCC1, KCC2, and NKCC1 mRNA expressions in the rat brain
Brain Res Dev Brain Res
The neuron-specific K-Cl cotransporter, KCC2Antibody development and initial characterization of the protein
J Biol Chem
A shared vesicular carrier allows synaptic corelease of GABA and glycine
Neuron
Control of postsynaptic Ca2+ influx in developing neocortex by excitatory and inhibitory neurotransmitters
Neuron
BDNF regulates spontaneous correlated activity at early developmental stages by increasing synaptogenesis and expression of the K+/Cl− co-transporter KCC2
Development
Development of the cerebellar system in relation to its evolutionstructureand functions
Cited by (40)
Development of the GABAergic network in the mouse spinal cord
2021, Factors Affecting Neurodevelopment: Genetics, Neurology, Behavior, and DietKCC2 regulates dendritic spine development
2020, Neuronal Chloride Transporters in Health and DiseaseEmbryonic development of GABAergic terminals in the mouse hypothalamic nuclei involved in feeding behavior
2018, Neuroscience ResearchChanges in the expression and localization of signaling molecules in mouse facial motor neurons during regeneration of facial nerves
2018, Journal of Chemical Neuroanatomy