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Activation of phospholipase C-γ1 in human keratinocytes by hyperosmolar shock without enzyme phosphorylation

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Abstract

Human keratinocytes are exposed to strong physical changes, and have the potentiality to react to external stimuli by switching on adaptation mechanisms. In hyperosmotically shocked keratinocytes a rapid and strong increase in calcium has been observed. We showed that this increase could not be prevented by growing the cells in medium devoid of calcium and in the presence of EGTA, indicating that the intracellular calcium increase was due to delivery from internal stores. Further, we observed an increased synthesis of dyacylglycerol and inositol trisphosphates after shock, suggesting that phospholipase C mediates both events. Our experiments demonstrated that osmotic shock in human keratinocytes leads to activation of phospholipase C-γ1, as measured using an in vitro assay system. This activation is independent of protein tyrosine phosphorylation and corresponded to a relocation of the enzyme to perinuclear membranes as shown by immunofluorescence.

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Fig. 1A, B
Fig. 2A–D
Fig. 3A, B
Fig. 4
Fig. 5

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Abbreviations

2-APB:

2-aminoethoxydiphenylborate

IP:

inositol phosphate

PA:

phosphatidic acid

PBS:

phosphate-buffered saline

PI-PLC:

phosphatidyl inositide-specific PLC

PIP:

phosphatidylinositolphosphate

PIP2 :

phosphatidylinositol(4,5)bisphosphate

PIP3 :

phosphatidylinositol(3,4,5)trisphosphate

PLC:

phospholipase C

PLD:

phospholipase D

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Authors and Affiliations

  1. Deutsches Krebsforschungszentrum, Im Neuenheimer Feld-242, 69120, Heidelberg, Germany

    I. Rodriguez, H. Cheng, K. Kabsch & A. Alonso

  2. Mannheim University of Applied Sciences, 68000, Mannheim, Germany

    A. Holloschi & M. Hafner

  3. Center of Pharmacology, University Hospital, 60590, Framkfurt, Germany

    M. Kaszkin

Authors
  1. I. Rodriguez
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  2. A. Holloschi
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  3. M. Kaszkin
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  4. H. Cheng
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  5. K. Kabsch
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  6. M. Hafner
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  7. A. Alonso
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Correspondence to A. Alonso.

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Rodriguez, I., Holloschi, A., Kaszkin, M. et al. Activation of phospholipase C-γ1 in human keratinocytes by hyperosmolar shock without enzyme phosphorylation. Arch Dermatol Res 295, 490–497 (2004). https://doi.org/10.1007/s00403-004-0457-4

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  • DOI: https://doi.org/10.1007/s00403-004-0457-4

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