Endoplasmic reticulum Ca2+ depletion activates XBP1 and controls terminal differentiation in keratinocytes and epidermis

Br J Dermatol. 2011 Jan;164(1):16-25. doi: 10.1111/j.1365-2133.2010.10046.x. Epub 2010 Nov 29.

Abstract

Background: Endoplasmic reticulum (ER) Ca(2+) depletion, previously shown to signal pathological stress responses, has more recently been found also to trigger homeostatic physiological processes such as differentiation. In keratinocytes and epidermis, terminal differentiation and barrier repair require physiological apoptosis and differentiation, as evidenced by protein synthesis, caspase 14 expression, lipid secretion and stratum corneum (SC) formation.

Objectives: To investigate the role of Ca(2+) depletion-induced ER stress in keratinocyte differentiation and barrier repair in vivo and in cell culture.

Methods: The SERCA2 Ca(2+) pump inhibitor thapsigargin (TG) was used to deplete ER calcium both in cultured keratinocytes and in mice. Levels of the ER stress factor XBP1, loricrin, caspase 14, lipid synthesis and intracellular Ca(2+) were compared after both TG treatment and barrier abrogation.

Results: We showed that these components of terminal differentiation and barrier repair were signalled by physiological ER stress, via release of stratum granulosum (SG) ER Ca(2+) stores. We first found that keratinocyte and epidermal ER Ca(2+) depletion activated the ER-stress-induced transcription factor XBP1. Next, we demonstrated that external barrier perturbation resulted in both intracellular Ca(2+) emptying and XBP1 activation. Finally, we showed that TG treatment of intact skin did not perturb the permeability barrier, yet stimulated and mimicked the physiological processes of barrier recovery.

Conclusions: This report is the first to quantify and localize ER Ca(2+) loss after barrier perturbation and show that homeostatic processes that restore barrier function in vivo can be reproduced solely by releasing ER Ca(2+), via induction of physiological ER stress.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Calcium / metabolism*
  • Caspase 14 / metabolism
  • Cell Differentiation / drug effects
  • Cell Differentiation / physiology*
  • Cells, Cultured
  • DNA-Binding Proteins / metabolism*
  • Endoplasmic Reticulum / drug effects
  • Endoplasmic Reticulum / metabolism*
  • Endoplasmic Reticulum / pathology
  • Enzyme Inhibitors / pharmacology
  • Epidermis / drug effects
  • Epidermis / metabolism*
  • Epidermis / pathology
  • Humans
  • Immunoblotting
  • Keratinocytes / cytology*
  • Keratinocytes / drug effects
  • Keratinocytes / pathology
  • Lipids / analysis
  • Membrane Proteins / metabolism
  • Mice
  • Polymerase Chain Reaction
  • Regulatory Factor X Transcription Factors
  • Thapsigargin / pharmacology
  • Transcription Factors / metabolism*
  • X-Box Binding Protein 1

Substances

  • DNA-Binding Proteins
  • Enzyme Inhibitors
  • Lipids
  • Membrane Proteins
  • Regulatory Factor X Transcription Factors
  • Transcription Factors
  • X-Box Binding Protein 1
  • XBP1 protein, human
  • Xbp1 protein, mouse
  • loricrin
  • Thapsigargin
  • Caspase 14
  • Calcium