Localization of sphingomyelin during the development of dorsal and tail epidermis of mice.
BACKGROUND
The water permeability barrier of the stratum corneum seems to be regulated primarily by lamellar bodies situated between the corneocytes; the lamellar bodies originate largely from polar lipid precursors, mainly sphingomyelin (SM), provided by the cells of the stratum granulosum via exocytosis of their lamellar body content.


OBJECTIVE
The aim of our study was to evaluate the cellular distribution of SM during development of the epidermis. Methods In this study, we investigated the expression and localization of SM in both adult and fetal mouse skin by a cytochemical detection method, immunofluorescence microscopy and immunoelectron microscopy, using anti-SM antibody, a specific binding protein to SM (lysenin), and Nile red stain. In addition, we measured transepidermal water loss to estimate the barrier function of the fetal skin.


RESULTS
We observed that SM was widely distributed from the basal layer to the granular layer in the adult mouse epidermis. An intense cytochemical reaction for SM was observed on embryonic day E14.5 of gestation just before the differentiation of the granular and squamous cells from the intermediate cells. The immunofluorescence indicating SM was detected in two regions, i.e. the most superficial zone of the granular layer and the upper spinous layer after the cell differentiation at the late gestational age. This distribution was not detected by conventional lipid staining, such as with Nile red stain. Immunoelectron microscopy revealed that SM was mainly localized in the intercellular spaces of the adult mouse epidermis and in the intracellular vesicles without a complete lamellar structure in the cytoplasm of epidermal cells of E14.5 fetuses. It is well known that the formation of the structurally mature cornified cell envelope occurs at E15.5 of development. The skin of fetuses at E16.5 showed a definite barrier function.


CONCLUSIONS
These findings suggest that SM dynamics is related to the formation of the lipid envelope, cell differentiation, and epidermal barrier function during development.
Yoshida
Y
Y
Department of Anatomy, Graduate School of Medicine, Kyoto University, Yoshidakonoe-cho, Sakyo-ku, Kyoto 606-8501, Japan. yyoshida@kuhp.kyoto-u.ac.jp
Yoneda
K
K
Umeda
M
M
Ide
C
C
Fujimoto
K
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D022162:Cytoplasmic Vesicles / Q000378:metabolism; D005314:Embryonic and Fetal Development / Q000502:physiology; D004817:Epidermis / Q000196:embryology / Q000254:growth & development* / Q000378:metabolism*; D005110:Extracellular Space / Q000378:metabolism; D005456:Fluorescent Dyes; D006801:Humans; D007091:Image Processing, Computer-Assisted; D051379:Mice; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D010078:Oxazines; D013109:Sphingomyelins / Q000276:immunology / Q000378:metabolism*; D013623:Tail; D014870:Water Loss, Insensible / Q000502:physiology
D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; D010078:Oxazines; D013109:Sphingomyelins; C044808:nile red