Original Article

Journal of Investigative Dermatology (1991) 96, 215–223; doi:10.1111/1523-1747.ep12461361

Membrane Structures in Normal and Essential Fatty Acid-Deficient Stratum Corneum: Characterization by Ruthenium Tetroxide Staining and X-Ray Diffraction

Sui Yuen E Hou1,2, Alok K Mitra3, Stephen H White4, Gopinathan K Menon2, Ruby Ghadially2 and Peter M Elias1,2

  1. 1Dermatology Service, Veterans Administration Medical Center, San Francisco, California, U.S.A.
  2. 2Departments of Dermatology, San Francisco, California, U.S.A.
  3. 3Biochemistry Biophysics, School of Medicine, University of California, San Francisco, California, U.S.A.
  4. 4Department of Physiology and Biophysics, College of Medicine, University of California, Irvine, California, U.S.A.

Received 9 May 1990; Accepted 21 September 1990.

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Abstract

Despite the importance of intercellular lamellar bilayers for stratum corneum (SC) barrier function, knowledge about the structure of these bilayers is limited due to their poor visualization and/or retention. Whereas substitution of ruthenium tetroxide (RuO4) for osmium tetroxide fixation provides clear images of these bilayers, the usefulness of RuO4 has been limited by its slow penetration and cytotoxicity. Utilizing a new fixation protocol for RuO4, we obtained clear images of lamellar domains at all levels of murine SC. Computer-aided image reconstructions demonstrated a lamellar spacing of 129 plusminus 2 Å, which agreed with x-ray diffraction data from parallel, unfixed samples (131 plusminus 2 Å), a spacing not affected by hydration. Furthermore, novel structures were seen in the intercellular spaces of normal SC. Finally, in murine essential fatty acid deficiency (EFAD), the overall lamellar spacing is comparable to normal [127 plusminus 7 Å by computer transform vs. 131.9 plusminus 2 Å (hydrated) and 129.6 plusminus 2.2 Å (dry) by x-ray diffraction]. Yet, these domains are structurally abnormal, displaying regions with either an excess or absence of lamellae. The new RuO4 protocol provides quantitative information about SC lamellar dimensions and morphologic abnormalities in bilayer distribution and substructure in EFAD stratum corneum that are not detected by either x-ray diffraction or computer-aided image reconstruction. Thus, the barrier abnormality in EFAD stratum corneum can be ascribed either to focal depletion of lamellae or abnormalities in lamellar substructure.

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