Formation of the Epidermal Calcium Gradient Coincides with Key Milestones of Barrier Ontogenesis in the Rodent Peter M. Elias, Patricia Nau, Karen Hanley,

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Formation of the Epidermal Calcium Gradient Coincides with Key Milestones of Barrier Ontogenesis in the Rodent Peter M. Elias, Patricia Nau, Karen Hanley, Chris Cullander, Debra Crumrine, Graham Bench, Elias Sideras-haddad, Theodora Mauro, Mary L. Williams, Kenneth R. Feingold Journal of Investigative Dermatology Volume 110, Issue 4, Pages 399-404 (April 1998) DOI: 10.1046/j.1523-1747.1998.00151.x Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Electron micrograph of 18 and 19 d old fetal mouse epidermis shows accelerated LB secretion. SG cytosol contains few LB, but the SG–SC interface is engorged (open arrows). After day 17–18, mature extracellular lamellae are present throughout the SC interstices (B, insert). Ruthenium tetroxide postfixation. Scale bars, 0.33 μm. Journal of Investigative Dermatology 1998 110, 399-404DOI: (10.1046/j.1523-1747.1998.00151.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Lanthanum still permeates on 17–18 d old, but not 19 d old fetal mouse epidermis. Direction of lanthanum nitrate perfusion is indicated by large arrows. Small arrows depict lanthanum in extracellular spaces of SC, and wavy arrows show lanthanum that has passed through to the outer surface of SC. Osmium tetroxide postfixation. Scale bars, 0.33 μm. Journal of Investigative Dermatology 1998 110, 399-404DOI: (10.1046/j.1523-1747.1998.00151.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Ion capture cytochemistry depicts a Ca++ gradient in 19 d old, but not 17 d old fetal mouse epidermis. (A) Day 17 epidermis displays incompletely developed SC, with evenly dispersed Ca++ precipitates in all epidermal layers, and high levels in the dermis (D). (B) In contrast, day 19 epidermis shows a distinct Ca++ gradient, with the highest Ca++ levels in the outer SG, lower levels in subjacent layers and SC. Osmium tetroxide postfixation. Scale bars, 0.33μm. Journal of Investigative Dermatology 1998 110, 399-404DOI: (10.1046/j.1523-1747.1998.00151.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 PIXEshows development of the Ca++ gradient in rat epidermis late in fetal development. A distinct gradient is present as early as day 20, with further acceleration thereafter. Data are mean ± SD for three separate recordings from three different samples. Journal of Investigative Dermatology 1998 110, 399-404DOI: (10.1046/j.1523-1747.1998.00151.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Potential relationship of Ca++ gradient to milestones of barrier development. Journal of Investigative Dermatology 1998 110, 399-404DOI: (10.1046/j.1523-1747.1998.00151.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions