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High resolution immunogold analysis reveals distinct subcellular compartmentation of protein kinase Cγ and δ in rat Purkinje cells

Cardell, M.; Landsend, A. S.; Eidet, J.; Wieloch, Tadeusz LU ; Blackstad, T. W. and Ottersen, O. P. (1997) In Neuroscience 82(3). p.709-725
Abstract

High resolution immunogold cytochemistry was used to investigate the subcellular distribution of protein kinase Cγ and δ in Purkinje cells of the rat cerebellum. Postembedding incubation with an antibody raised to a peptide sequence near the C-terminus of protein kinase Cγ resulted in strong labelling along the dendrosomatic plasma membrane. A quantitative analysis indicated that this labelling reflected the existence of two pools of protein kinase Cγ; one membrane associated pool and one cytoplasmic pool located within 50 nm of the plasma membrane. The labelling along the plasma membrane showed a pronounced and abrupt increase when moving from the cell body into the axon initial segment. Gold particles signalling protein kinase Cγ were... (More)

High resolution immunogold cytochemistry was used to investigate the subcellular distribution of protein kinase Cγ and δ in Purkinje cells of the rat cerebellum. Postembedding incubation with an antibody raised to a peptide sequence near the C-terminus of protein kinase Cγ resulted in strong labelling along the dendrosomatic plasma membrane. A quantitative analysis indicated that this labelling reflected the existence of two pools of protein kinase Cγ; one membrane associated pool and one cytoplasmic pool located within 50 nm of the plasma membrane. The labelling along the plasma membrane showed a pronounced and abrupt increase when moving from the cell body into the axon initial segment. Gold particles signalling protein kinase Cγ were also enriched in putative Purkinje axon terminals in the dentate nucleus. The only organelle showing a consistent immunolabelling for protein kinase Cγ was the Golgi apparatus where the gold particles were restricted to the trans face. Protein kinase Cγ immunoreactivity also occurred in the Purkinje cell spines, with an enrichment in or near the postsynaptic density. Antibodies to protein kinase Cδ produced a very different labelling pattern in the Purkinje cells. Most of the gold particles were associated with rough endoplasmic reticulum, particularly with those cisternae that were located close to the nucleus or in the nuclear indentations. No significant protein kinase C5 immunolabelling was detected at the plasma membrane or in Purkinje cell spines. The present data point to a highly specific compartmentation of the two major protein kinase C isozyme in Purkinje cells and suggest that these isozymes act on different substrates and hence have different regulatory functions within these neurons.

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author
organization
publishing date
type
Contribution to journal
publication status
published
keywords
Freeze substitution, Golgi apparatus, Immunogold cytochemistry, Initial segment, Long term depression, Protein kinase
in
Neuroscience
volume
82
issue
3
pages
709 - 725
publisher
Elsevier
external identifiers
  • Scopus:0343852716
ISSN
0306-4522
DOI
10.1016/S0306-4522(97)00305-9
language
English
LU publication?
yes
id
31b0b1e4-2784-4128-a9d5-73239defad5a
date added to LUP
2016-10-05 16:08:49
date last changed
2016-11-28 16:07:47
@misc{31b0b1e4-2784-4128-a9d5-73239defad5a,
  abstract     = {<p>High resolution immunogold cytochemistry was used to investigate the subcellular distribution of protein kinase Cγ and δ in Purkinje cells of the rat cerebellum. Postembedding incubation with an antibody raised to a peptide sequence near the C-terminus of protein kinase Cγ resulted in strong labelling along the dendrosomatic plasma membrane. A quantitative analysis indicated that this labelling reflected the existence of two pools of protein kinase Cγ; one membrane associated pool and one cytoplasmic pool located within 50 nm of the plasma membrane. The labelling along the plasma membrane showed a pronounced and abrupt increase when moving from the cell body into the axon initial segment. Gold particles signalling protein kinase Cγ were also enriched in putative Purkinje axon terminals in the dentate nucleus. The only organelle showing a consistent immunolabelling for protein kinase Cγ was the Golgi apparatus where the gold particles were restricted to the trans face. Protein kinase Cγ immunoreactivity also occurred in the Purkinje cell spines, with an enrichment in or near the postsynaptic density. Antibodies to protein kinase Cδ produced a very different labelling pattern in the Purkinje cells. Most of the gold particles were associated with rough endoplasmic reticulum, particularly with those cisternae that were located close to the nucleus or in the nuclear indentations. No significant protein kinase C5 immunolabelling was detected at the plasma membrane or in Purkinje cell spines. The present data point to a highly specific compartmentation of the two major protein kinase C isozyme in Purkinje cells and suggest that these isozymes act on different substrates and hence have different regulatory functions within these neurons.</p>},
  author       = {Cardell, M. and Landsend, A. S. and Eidet, J. and Wieloch, Tadeusz and Blackstad, T. W. and Ottersen, O. P.},
  issn         = {0306-4522},
  keyword      = {Freeze substitution,Golgi apparatus,Immunogold cytochemistry,Initial segment,Long term depression,Protein kinase},
  language     = {eng},
  month        = {10},
  number       = {3},
  pages        = {709--725},
  publisher    = {ARRAY(0xeb8e040)},
  series       = {Neuroscience},
  title        = {High resolution immunogold analysis reveals distinct subcellular compartmentation of protein kinase Cγ and δ in rat Purkinje cells},
  url          = {http://dx.doi.org/10.1016/S0306-4522(97)00305-9},
  volume       = {82},
  year         = {1997},
}