Origin of the Epidermal Calcium Gradient: Regulation by Barrier Status and Role of Active vs Passive Mechanisms Peter M. Elias, MD., Barbara E. Brown,

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Origin of the Epidermal Calcium Gradient: Regulation by Barrier Status and Role of Active vs Passive Mechanisms Peter M. Elias, MD., Barbara E. Brown, Debra Crumrine, Kenneth R. Feingold Journal of Investigative Dermatology Volume 119, Issue 6, Pages 1269-1274 (December 2002) DOI: 10.1046/j.1523-1747.2002.19622.x Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Reappearance of Ca gradient is accelerated when barrier function is artificially restored. (A) Normal (Nm) epidermal Ca gradient, with highest density of electron-dense precipitates in SG, and low levels in both SC, and subjacent stratum spinosum (SS) and stratum basale (SB). (B) Three hours after tape-stripping (TS; note removal of most SC), little or no Ca is present in epidermis, although Ca precipitates can be seen in the dermis (D). (C,D) When barrier is artificially restored, immediately after tape-stripping by application of either Gore-Tex® (G) or petrolatum (Vaseline, V) wrap, substantial Ca precipitates are present in epidermis. Scale bars=2 μm. Pyroantimonate precipitation, followed by OsO4 postfixation. Journal of Investigative Dermatology 2002 119, 1269-1274DOI: (10.1046/j.1523-1747.2002.19622.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Reappearance of Ca gradient is delayed when barrier recovery is delayed. (A) Six hours after tape-stripping (TS) and a single lovastatin (Lov) application, little or no Ca precipitates are evident in epidermis. (B) In contrast, in TS plus vehicle (Veh)-treated sites, substantial Ca has returned (arrows). Scale bar=1 μm. Pyroantimonate precipitation, followed by OsO4 postfixation. Journal of Investigative Dermatology 2002 119, 1269-1274DOI: (10.1046/j.1523-1747.2002.19622.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Barrier recovery is delayed at 4°C, with or without occlusion. Sets of normal hairless mice were tape-stripped until TEWL > 4 mg per cm2 per h, and either allowed to recover at room temperature, at 4°C by immersion in iced normal saline (NS), exposure to cold air (4–10°C), or occluded with Gore-Tex®, followed by application of iced NS. Data shown represent mean±SEM of the percent barrier recovery at 5 h (n=5 or 6 for each group). Journal of Investigative Dermatology 2002 119, 1269-1274DOI: (10.1046/j.1523-1747.2002.19622.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Artificial restoration of barrier accelerates return of Ca gradient, even at 4°C. (A) Five hours after tape-stripping (TS) and air exposure at 23°C, substantial Ca has returned to epidermis. (B) Immediately after TS and prior to exposure to cold air (Pre: –4°C), the Ca gradient is absent; i.e., levels of precipitate in SG and stratum spinosum (SS) are very low. A few precipitates are present in the basal layer (SB). In contrast, tape-stripped epidermis, maintained at 4–10°C (cold air) reveals substantial return of Ca to epidermis, with a few precipitates reaching the SC, but no gradient formation. (C) In contrast, tape-stripped, cold-exposed sites, occluded with Gore-Tex® (GT) reveal a substantial return of the Ca gradient; i.e., higher levels in the SG than in the basal/spinous layers. Scale bars: (A) 2 μm, (B–D) 1.0 μm. Pyroantimonate precipitation, followed by OsO4 postfixation. Journal of Investigative Dermatology 2002 119, 1269-1274DOI: (10.1046/j.1523-1747.2002.19622.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Pathophysiologic implications of a barrier-dominated Ca gradient. Journal of Investigative Dermatology 2002 119, 1269-1274DOI: (10.1046/j.1523-1747.2002.19622.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions