Expression of the protein kinase D (PKD) family during mouse embryogenesis

ABSTRACT. Protein Kinase D (PKD) 1, 2, and 3 are members of the PKD family. PKDs influence many cellular processes, including cell polarity, structure of the Golgi, polarized transport from the Golgi to the basolateral plasma membrane, and actin polymerization. However, the role of the PKD family in cell polarity has not yet been elucidated in vivo. Here, we show that KO mice displayed similar localization of the apical and basolateral proteins, transport of VSV-G and a GPI-anchored protein, and similar localization of actin filaments. As DKO mice were embryonic lethal, we generated MEFs that lacked all PKD isoforms from the PKD1 and PKD2 double floxed mice using Cre recombinase and PKD3 siRNA. We observed a similar localization of various organelles, a similar time course in the transport of VSV-G and a GPI-anchored protein, and a similar distribution of F-actin in the PKD-null MEFs. Collectively, our results demonstrate that the complete deletion of PKDs does not affect the transport of VSV-G or a GPI-anchored protein, and the distribution of F-actin. However, simultaneous deletion of PKD1 and PKD2 affect embryonic development, demonstrating their functional redundancy during development.

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“…2F). These results largely reflected the distribution of the mRNA observed in a previous study (Oster et al, 2006).…”

Section: Fig 2 Generation Of Pkd2 Ko (Pkd2supporting

confidence: 86%

The Role of PKD in Cell Polarity, Biosynthetic Pathways, and Organelle/F-actin Distribution

Atik

Kunii

Avriyanti

et al. 2014

Cell Struct. Funct.

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“…Because PKD1 is expressed in numerous tissues (20), we used the Cre-loxP recombination system to generate a conditional PKD1 (Prkcm) allele that could be deleted specifically in cardiomyocytes. LoxP sites were inserted into the Prkcm locus to flank exons 12 through 14, which encode part of the catalytic domain of PKD1, including the ATP binding motif that is essential for kinase function (Fig.…”

Section: Resultsmentioning

confidence: 99%

Requirement of protein kinase D1 for pathological cardiac remodeling

Fielitz

Kim

Shelton

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

220

244

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The adult heart responds to biomechanical stress and neurohormonal signaling by hypertrophic growth, accompanied by fibrosis, diminished pump function, and activation of a fetal gene program. Class II histone deacetylases (HDACs) suppress stress-dependent remodeling of the heart via their association with the MEF2 transcription factor, an activator of heart disease. Protein kinase D (PKD) is a stress-responsive kinase that phosphorylates class II HDACs, resulting in their dissociation from MEF2 with consequent activation of MEF2 target genes. To test whether PKD1 is required for pathological cardiac remodeling in vivo, we generated mice with a conditional PKD1-null allele. Mice with cardiac-specific deletion of PKD1 were viable and showed diminished hypertrophy, fibrosis, and fetal gene activation as well as improved cardiac function in response to pressure overload or chronic adrenergic and angiotensin II signaling. We conclude that PKD1 functions as a key transducer of stress stimuli involved in pathological cardiac remodeling in vivo.cardiac hypertrophy ͉ histone deacetylase ͉ stress-responsive kinase

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“…All three PKD isoforms are expressed in the CNS already at embryonic stages in mice (Oster et al, 2006;Ellwanger et al, 2008). PKD activity has been recently implicated in regulating early neuronal polarization (Yin et al, 2008), early dendritic development (Horton et al, 2005), or selective transport and endocytosis of dendritic proteins (Bisbal et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

“…Furthermore, studies in polarized epithelial cells revealed that PKD is selectively involved in basolateral membrane protein transport (Yeaman et al, 2004). PKD was also reported to regulate lipid transfer between the endoplasmic reticulum and the Golgi apparatus (Fugmann et al, 2007).All three PKD isoforms are expressed in the CNS already at embryonic stages in mice (Oster et al, 2006;Ellwanger et al, 2008). PKD activity has been recently implicated in regulating early neuronal polarization (Yin et al, 2008), early dendritic development (Horton et al, 2005), or selective transport and endocytosis of dendritic proteins (Bisbal et al, 2008).…”

mentioning

confidence: 99%

Protein Kinase D Controls the Integrity of Golgi Apparatus and the Maintenance of Dendritic Arborization in Hippocampal Neurons

Czöndör

Ellwanger²,

Fuchs³

et al. 2009

MBoC

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Protein kinase D (PKD) is known to participate in various cellular functions, including secretory vesicle fission from theGolgi 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. INTRODUCTIONNeurons are nondividing and extremely polarized cells, with enormous membrane surface compared with the size of the soma. These features assume a highly specialized secretory machinery, which resembles in certain parts the directed transport mechanisms described in polarized nonneuronal cells (Winckler and Mellman, 1999;Horton and Ehlers, 2004). The vast neuronal membrane surface is functionally and structurally divided into axonal and somatodendritic compartments, possessing specific lipid and protein components required for the spatially different functions, e.g., for pre-or postsynaptic activity (Dresbach et al., 2001;Sheng, 2001;Bresler et al., 2004). The constitutive and precisely directed supply of surface membrane components is indispensable for the function of neurons, especially when considering that postmitotic neurons normally serve throughout the life span of the organism. Up to now, we are far from understanding how the extreme polarization has been evolved and is maintained in neurons.Despite a likely central role in neuronal morphogenesis and membrane trafficking, little is known about the special structure and transport features of the neuronal Golgi apparatus. A thread-like and reticular structure of the Golgi apparatus has been already described in many neuronal types (Takamine et al., 2000;Fujita and Okamoto, 2005;Horton et al., 2005). Central neuronal Golgi complex is localized in the soma and often extends into the principal dendrites, but Golgi elements were also found as discontinuous structures in the distal dendrites, often near synaptic contacts or in dendritic spines (Gardiol et al., 1999;Pierc...

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Expression of the protein kinase D (PKD) family during mouse embryogenesis

Cited by 31 publications

References 26 publications

The Role of PKD in Cell Polarity, Biosynthetic Pathways, and Organelle/F-actin Distribution

The Role of PKD in Cell Polarity, Biosynthetic Pathways, and Organelle/F-actin Distribution

Requirement of protein kinase D1 for pathological cardiac remodeling

Protein Kinase D Controls the Integrity of Golgi Apparatus and the Maintenance of Dendritic Arborization in Hippocampal Neurons

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