Acute barrier perturbation abolishes the Ca2+ and K+ gradients in murine epidermis: quantitative measurement using PIXE.

Epidermal permeability barrier homeostasis requires the delivery of lipids and hydrolytic enzymes by lamellar body exocytosis from the uppermost granular cells, a process that is upregulated following barrier disruption. As lamellar body secretion is controlled by ionic concentrations, especially Ca2+ and K+, we used a quantitative technique, microbeam proton-induced X-ray emission, to measure Ca2+, K+, Cl-, and P concentrations before and after acute barrier perturbation by acetone applications. We found a steep gradient of Ca2+ in normal tissue, peaking in the outer stratum granulosum, which disappeared after barrier disruption, and partially reformed as the barrier recovered. A similar gradient, peaking somewhat lower in the epidermis (i.e., at the stratum granulosum-stratum corneum interface), was found for K+. Epidermal concentrations of K+ also decreased after barrier abrogation, although to a lesser extent than Ca2+. In contrast, P and Cl- demonstrated distribution gradients at baseline, which remained unchanged after barrier disruption. These studies quantitate the levels of Ca2+, K+, Cl-, and P within specific epidermal cell layers at baseline, and in relation to changes in permeability barrier integrity. Ca2+ and K+, but not Cl- or P, decrease after barrier disruption, consistent with these two ion's role in barrier repair.