1. ¹Department of Human Anatomy and Cell Biology, University of Liverpool, Liverpool, UK
2. ²Department of Clinical Biochemistry, University of Liverpool, Liverpool, UK
3. ³Department of Dermatology, Broadgreen Hospital, Liverpool, UK

Correspondence: Dr Helen E. Burrell, Biomolecular Sciences, James Parsons Building, Liverpool John Moores University, Byrom Street, Liverpool L3 3AF, UK. E-mail: H.Burrell@ljmu.ac.uk

⁴Current address: Biomolecular Sciences, James Parsons Building, Liverpool John Moores University, Byrom Street, Liverpool L3 3AF, UK

Received 2 August 2006; Revised 22 August 2007; Accepted 30 August 2007; Published online 15 November 2007.

Abstract

In the epidermis, local and systemic factors including extracellular nucleotides and parathyroid hormone-related protein (PTHrP) regulate keratinocyte proliferation and differentiation. Extracellular nucleotides increase proliferation via activation of P2 receptors and induction of calcium transients, while endoproteases cleave PTHrP, resulting in fragments with different cellular functions. We investigated the effects of adenosine 5'-triphosphate (ATP) alone and in combination with synthetic PTHrP peptides on calcium transients in HaCaT cells. ATP induced calcium transients, while PTHrP peptides did not. C-terminal and mid-molecule PTHrP peptides (1–100 pM) potentiated ATP-induced calcium transients independently of calcium influx. 3-isobutyl-1-methylxanthine potentiated ATP-induced calcium transients, suggesting that a cyclic monophosphate is responsible. Cyclic AMP is not involved, but cyclic GMP is a likely candidate since the protein kinase G inhibitor, KT5823, inhibited potentiation. Co-stimulation with ATP and either PTHrP (43–52) or PTHrP (70–77) increased proliferation, suggesting that this is important in the regulation of cell turnover and wound healing and may be a mechanism for hyperproliferation in skin disorders such as psoriasis. Finally, PTHrP fragments potentiated bradykinin-induced calcium transients, suggesting a role in inflammation in the skin. Since PTHrP is found in many normal and malignant cells, potentiation is likely to have a wider role in modulating signal transduction events.