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PMID:19211839

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Citation

Czöndör, K, Ellwanger, K, Fuchs, YF, Lutz, S, Gulyás, M, Mansuy, IM, Hausser, A, Pfizenmaier, K and Schlett, K (2009) Protein kinase D controls the integrity of Golgi apparatus and the maintenance of dendritic arborization in hippocampal neurons. Mol. Biol. Cell 20:2108-20

Abstract

Protein kinase D (PKD) is known to participate in various cellular functions, including secretory vesicle fission from the Golgi and plasma membrane-directed transport. Here, we report on expression and function of PKD in hippocampal neurons. Expression of an enhanced green fluorescent protein (EGFP)-tagged PKD activity reporter in mouse embryonal hippocampal neurons revealed high endogenous PKD activity at the Golgi complex and in the dendrites, whereas PKD activity was excluded from the axon in parallel with axonal maturation. Expression of fluorescently tagged wild-type PKD1 and constitutively active PKD1(S738/742E) (caPKD1) in neurons revealed that both proteins were slightly enriched at the trans-Golgi network (TGN) and did not interfere with its thread-like morphology. By contrast, expression of dominant-negative kinase inactive PKD1(K612W) (kdPKD1) led to the disruption of the neuronal Golgi complex, with kdPKD1 strongly localized to the TGN fragments. Similar findings were obtained from transgenic mice with inducible, neuron-specific expression of kdPKD1-EGFP. As a prominent consequence of kdPKD1 expression, the dendritic tree of transfected neurons was reduced, whereas caPKD1 increased dendritic arborization. Our results thus provide direct evidence that PKD activity is selectively involved in the maintenance of dendritic arborization and Golgi structure of hippocampal neurons.

Links

PubMed PMC2663930 Online version:10.1091/mbc.E08-09-0957

Keywords

Animals; Cell Compartmentation/drug effects; Cell Differentiation/drug effects; Cell Polarity/drug effects; Dendrites/drug effects; Dendrites/enzymology; Doxycycline/pharmacology; Enzyme Activation/drug effects; Genes, Dominant; Genes, Reporter; Golgi Apparatus/drug effects; Golgi Apparatus/enzymology; Green Fluorescent Proteins/metabolism; Hippocampus/cytology; Humans; Mice; Neurons/cytology; Neurons/drug effects; Neurons/enzymology; Protein Kinase C/metabolism; Protein Transport/drug effects; Recombinant Fusion Proteins/metabolism; Transfection; trans-Golgi Network/drug effects; trans-Golgi Network/enzymology

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:KPCD1

located_in

GO:0005802: trans-Golgi network

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:KPCD1

involved_in

GO:0007030: Golgi organization

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:KPCD1

part_of

GO:0005802: trans-Golgi network

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:KPCD1

involved_in

GO:0010976: positive regulation of neuron projection development

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

See also

References

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