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The effect of extracellular calcium concentration on calcium-mediated cell signaling in NF1 tumor suppressor-deficient keratinocytes

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Abstract

Capacitative calcium entry and calcium wave propagation were studied in keratinocytes from healthy volunteers and patients with type 1 neurofibromatosis (NF1) in calcium-depleted and in low calcium culture medium. In previous studies, we found evidence that mutations of the NF1 tumor suppressor gene can lead to altered calcium-mediated cell signaling in keratinocytes cultured in the presence of a high extracellular calcium concentration. The present study demonstrated that the differences between normal and NF1 keratinocytes were dependent on extracellular calcium concentration. Specifically, when keratinocytes were exposed to thapsigargin under calcium-depleted culture conditions the subsequent increase in free intracellular calcium concentration was moderate in NF1 keratinocytes compared to controls. The finding indicates lowered endoplasmic calcium stores in NF1 which may also in part explain the reduced activation signal for capacitative calcium influx and the wound-induced intracellular Ca2+ transient observed in NF1 keratinocytes maintained in culture medium containing 0.05 mM calcium. The differences between control and NF1 keratinocytes were most pronounced when the cells were cultured in the presence of a high (1.8 mM) calcium concentration. Since elevated extracellular calcium levels induce keratinocytes to form cellular contacts and lead to terminal differentiation, markedly aberrant responses of NF1 keratinocytes in the presence of a high calcium concentration may help to explain previous findings on impaired formation of cellular junctions and differentiation in NF1 deficient cells.

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Acknowledgements

This work was supported by grants from Finnish Cancer Societies, Academy of Finland, Turku University Foundation, Oulu University Hospital (grant H01139), Turku University Central Hospital (grants 11606 and 13338), Paulo Foundation, Emil Aaltonen Foundation, Research Foundation of Farmos, The Cancer Society of Northern Finland, The Finnish Medical Society Duodecim, Finnish Cultural Foundation, Tyyni Tani Foundation, Foundation for Memory of Maud Kuistila, Nona and Kullervo Väre Foundation/Ester Mäkelä Foundation, the Finnish Medical Foundation and the K. Albin Johansson Foundation. We thank Drs. Hannu Rajaniemi, Juha Tuukkanen and Vesa Aaltonen for fruitful discussions. We thank Risto Bloigu, Mika Kihlström, Liisa Kärki, Seija Leskelä and Eero Oja for expert technical assistance.

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Authors and Affiliations

  1. Department of Anatomy and Cell Biology, University of Oulu, PB 5000, 90014, Finland

    Timo Korkiamäki, Heli Ylä-Outinen, Pekka Leinonen, Jussi Koivunen & Juha Peltonen

  2. Department of Dermatology, University of Oulu, Oulu, Finland

    Juha Peltonen

  3. Department of Anatomy, University of Turku, Turku, Finland

    Juha Peltonen

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  1. Timo Korkiamäki
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  2. Heli Ylä-Outinen
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Correspondence to Timo Korkiamäki.

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Authors’ contributions

Timo Korkiamäki study design, data collection, statistical analysis, data interpretation, manuscript preparation, literature search, collection of funds; Heli Ylä-Outinen study design, data collection, data interpretation, manuscript preparation, collection of funds; Pekka Leinonen data interpretation, manuscript preparation; Jussi Koivunen study design, data interpretation, collection of funds; Juha Peltonen study design, data interpretation, manuscript preparation, collection of funds.

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Korkiamäki, T., Ylä-Outinen, H., Leinonen, P. et al. The effect of extracellular calcium concentration on calcium-mediated cell signaling in NF1 tumor suppressor-deficient keratinocytes. Arch Dermatol Res 296, 465–472 (2005). https://doi.org/10.1007/s00403-004-0538-4

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