High resolution immunogold analysis reveals distinct subcellular compartmentation of protein kinase Cγ and δ in rat Purkinje cells
(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.
(Less)
- author
- Cardell, M. ; Landsend, A. S. ; Eidet, J. ; Wieloch, Tadeusz LU ; Blackstad, T. W. and Ottersen, O. P.
- organization
- publishing date
- 1997-10-24
- 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
-
- pmid:9483530
- 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
- 2022-01-30 06:35:02
@article{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}},
keywords = {{Freeze substitution; Golgi apparatus; Immunogold cytochemistry; Initial segment; Long term depression; Protein kinase}},
language = {{eng}},
month = {{10}},
number = {{3}},
pages = {{709--725}},
publisher = {{Elsevier}},
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}},
doi = {{10.1016/S0306-4522(97)00305-9}},
volume = {{82}},
year = {{1997}},
}