Skin barrier function is conferred by the outer layer of epidermis, the stratum corneum, and is essential for terrestrial life. Quantitative trans-epidermal water loss assays show that barrier forms late in embryogenesis, permitting the foetus to survive a terrestrial environment at birth. Using qualitative in situ assays for skin permeability, we show that barrier forms in a patterned manner late in mouse gestation. Barrier forms at specific epidermal sites, then spreads around the embryo as a moving front. The moving front of permeability change is accompanied by multiple changes in the outer, stratum corneum-precursor cells. We use the permeability assays to show that final stages of cornified envelope assembly are coordinated with initial stages of barrier formation. Hence the whole-mount permeability assays record developmental acquisition of a known, essential component of the adult barrier. We demonstrate the authenticity of the whole-mount assays after maternal glucocorticoid therapy (known to accelerate barrier formation) and in additional species including the rat where barrier formation is well characterized by TEWL assay (Aszterbaum, M., Menon, G. K., Feingold, K. R. and Williams, M. L. Pediatr. Res. 31, 308–317). The demonstration of patterned barrier formation in other species suggests patterned change as the universal mode of embryonic barrier acquisition. These results highlight the importance of patterning as a mode of epidermal maturation during development.

REFERENCES

Aszterbaum
M.
,
Feingold
K. R.
,
Menon
G. K.
,
Williams
M. L.
(
1993
)
Glucocorticoids accelerate fetal maturation of the epidermal permeability barrier in the rat.
J. Clin. Invest
91
,
2703
2708
Aszterbaum
M.
,
Menon
G. K.
,
Feingold
K. R.
,
Williams
M. L.
(
1992
)
Ontogeny of the epidermal barrier to water loss in the rat: correlation of function with stratum corneum structure and lipid content.
Pediatr. Res
31
,
308
317
Bickenbach
J. R.
,
Greer
J. M.
,
Bundman
D. S.
,
Rothnagel
J. A.
,
Roop
D. R.
(
1995
)
Loricrin expression is coordinated with other epidermal proteins and the appearance of lipid lamellar granules in development.
J. Invest. Dermatol
104
,
405
410
Blank
I. H.
(
1953
)
Further observations on factors which influence the water content of the stratum corneum.
J.Invest. Dermatol
21
,
259
269
Byrne
C.
,
Tainsky
M.
,
Fuchs
E.
(
1994
)
Programming gene expression in developing epidermis.
Development
120
,
2369
2383
Downing
D. T.
(
1992
)
Lipid and protein structures in the permeability barrier of mammalian epidermis.
J. Lipid Res
33
,
301
313
Elias
P. M.
(
1983
)
Epidermal lipids, barrier function and desquamation.
J. Invest. Dermatol
80
,
44
49
Elias
P. M.
,
Menon
G. K.
(
1991
)
Structural and lipid biochemical correlates of the epidermal permeability barrier.
Adv. Lipid Res
24
,
1
26
Evans
N.
,
Rutter
N.
(
1986
)
Development of the epidermis in the newborn.
Biol. Neonate
49
,
74
80
Hammarlund
K.
,
Sedin
G.
(
1979
).
Transepidermal water loss in newborn.III. Relationship to gestational age.
Acta Paediatr. Scand
68
,
795
801
Hanley
K.
,
Jiam
U.
,
Holleran
W.M.
,
Elias
P.M.
,
Williams
M.L.
,
Feingold
K.R.
(
1997
)
Glucosylceramide metabolism is regulated duringnormal and hormonally-stimulated epidermal barrier development in the rat.
J. Lipid Res
38
,
576
584
Hanley
K.
,
Rassner
U.
,
Elias
P. M.
,
Williams
M. L.
,
Feingold
K. R.
(
1996
)
Epidermal barrier ontogenesis: maturation in serum-free media and acceleration by glucocorticoids and thyroid hormone but not selected growth factors.
J. Invest. Dermatol
106
,
404
411
Hanley
K.
,
Rassner
U.
,
Jiang
Y.
,
Vansomphone
D.
,
Crumbine
D.
,
Komuves
L.
,
Elias
P. M.
,
Feingold
K. R.
,
Williams
M. L.
(
1996
)
Hormonal basis for the gender difference in epidermal barrier formation in the fetal rat.
J. Clin. Invest
97
,
2576
2584
Hanson
J.
(
1947
)
The histogenesis of the epidermis in the rat and mouse.
J. Anat
81
,
174
197
Hayward
A. F.
(
1983
)
The permeability of the epithelium of the skin of fetal rats demonstrated with a lanthanum-containing solution.
J. Anat
136
,
379
388
Hohl
D.
,
Mehrel
T.
,
Lichti
U.
,
Turner
M.L.
,
Roop
D.R.
,
Steinert
P.M.
(
1991
)
Characterization of human loricrin.
J. Biol. Chem
266
,
6626
6636
Holbrook
K. A.
,
Odland
G. F.
(
1980
)
Regional development of the human epidermis in the first trimester embryo and the second trimester fetus (ages related to the timing of amniocentesis and fetal biopsy).
J. Invest. Dermatol
80
,
161
168
Huber
M.
,
Rettler
I.
,
Bernasconi
K.
,
Frenk
E.
,
Lavrijsen
S.
,
Ponec
M.
,
Bon
A.
,
Lautenschlager
S.
,
Schorderet
D.
,
Hohl
D.
(
1995
)
Mutations of keratinocyte transglutaminase in lamellar ichthyosis.
Science
267
,
525
528
Imakado
S.
,
Bickenbach
J.
,
Bundman
D. S.
,
Rothnagel
J. A.
,
Attar
P. S.
,
Wang
X.-J.
,
Walczak
V. R.
,
Wisniewski
S.
,
Pote
J.
,
Gordon
J. S.
,
Heyman
R. A.
,
Evans
R. A.
,
Roop
D. R.
(
1995
)
Targeting expression of a dominant-negative retinoic acid receptor mutant in the epidermis of transgenic mice results in loss of barrier function.
Genes Dev
9
,
317
329
Ishida-Yamamoto
A.
,
Eady
R. A. J.
,
Watt
F. M.
,
Roop
D. R.
,
Hohl
D.
,
Iizuka
H.
(
1996
)
Immunoelectron microscopic analysis of cornified cell envelope formation in normal and psoriatic epidermis.
J. Histochem. Cytochem
44
,
165
175
Kitraki
E.
,
Kittas
C.
,
Stylianopoulou
F.
(
1997
)
Glucocorticoid receptor gene expression during rat embryogenesis.
Differentiation
62
,
21
31
Lazo
N. D.
,
Meine
J. G.
,
Downing
D. T.
(
1995
)
Lipids are covalently attached to rigid corneocyte protein envelopes existing predominantly as-sheets: A solid-state nuclear magnetic resonance study.
J. Invest. Dermatol
105
,
296
300
Maestrini
E.
,
Monaco
A. P.
,
McGrath
J. A.
,
Ishida-Yamamoto
A.
,
Camisa
C.
,
Hovnanian
A.
,
Weeks
D. E.
,
Lathrop
M.
,
Uitto
J.
,
Christiano
A. M.
(
1996
)
A molecular defect in loricrin, the major component of the cornified cell envelope, underlies Vorwinkel's syndrome.
Nature Genet
13
,
70
77
Manabe
M.
,
Sanchez
M.
,
Sun
T. T.
,
Dale
B. A.
(
1991
)
Interaction of filaggrin with keratin filaments during advanced stages of normal human epidermal differentiation in Ichthyosis vulgaris.
Differentiation
48
,
43
50
Mehrel
T.
,
Hohl
D.
,
Rothnagel
J. A.
,
Longley
M. A.
,
Bundman
D.
,
Cheng
C.
,
Lichti
U.
,
Bisher
M. E.
,
Steven
A. C.
,
Steinert
P. M.
,
Yuspa
S. H.
,
Roop
D. R.
(
1990
)
Identification of a major keratinocyte cell envelope protein, loricrin.
Cell
61
,
1103
1112
Michel
S.
,
Schmidt
R.
,
Schroot
B.
,
Reichert
U.
(
1988
)
Morphologicaland biochemical characterization of the cornified envelopes from human epidermal keratinocytes of different origin.
J. Invest. Dermatol
91
,
11
15
Newman
G. R.
,
Hobot
J. A.
(
1987
)
Modern acrylics for post-embedding immunostaining techniques.
J. Histochem. Cytochem
35
,
971
981
Nilsson
G. E.
(
1977
)
Measurement of water exchange through the skin.
Med. & Biol. Eng. Comput
15
,
209
218
Okah
F. A.
,
Pickens
W. L.
,
Hoath
S. B.
(
1995
)
Effect of prenatal steroids on skin surface hydrophobicity in the premature rat.
Pediatr. Res
37
,
402
408
Robertson
D.
,
Monaghan
P.
,
Clarke
C.
,
Atherton
A. J.
(
1992
)
An appraisal of low-temperature embedding by progressive lowering of temperature into Lowicryl HM20 for immunocytochemical studies.
J. Microscopy
168
,
85
100
Roop
D.
(
1995
)
Defects in the barrier.
Science
267
,
474
–.
Russell
L. J.
,
DiGiovanna
J. J.
,
Rogers
G. R.
,
Steinert
P. M.
,
Hashem
N.
,
Compton
J. G.
,
Bale
S. J.
(
1995
)
Mutations in the gene for transglutaminase 1 in autosomal recessive lamellar ichthyosis.
Nature Genetics
9
,
279
283
Sengel
P.
(
1990
)
Pattern formation in skin development.
Int. J. Dev. Biol
34
,
33
50
Sicard
R. E.
,
Werner
J. C.
(
1992
)
Dexamethasone induces a transient relative cardiomegaly in neonatal rats.
Pediatr. Res
31
,
359
363
Steinert
P. M.
,
Marekov
L. N.
(
1995
)
The proteins elafin, filaggrin, keratin intermediate filaments, loricrin, and small proline-rich proteins 1 and 2 are isodipeptide cross-linked components of the human epidermal cornified cell envelope.
J. Biol. Chem
270
,
17
–.
Steinert
P. M.
,
Marekov
L. N.
(
1997
)
Direct evidence that involucrin is a major early isopeptide cross-linked component of the keratinocyte cornified cell envelope.
J. Biol. Chem
272
,
2021
2030
Steven
A. C.
,
Steinert
P. M.
(
1994
)
Protein composition of cornified cell envelopes of epidermal keratinocytes.
J. Cell Sci
107
,
693
700
Steven
A. C.
,
Bisher
M. E.
,
Roop
D.
,
Steinert
P. M.
(
1990
)
Biosynthetic pathways of filaggrin and loricrin-two major proteins expressed by terminally differentiated epidermal keratinocytes.
J. Struct. Biol
104
,
150
162
Sun
T.-T.
,
Green
H.
(
1976
)
Differentiation of the epidermal keratinocyte in cell culture: formation of the cornified envelope.
Cell
9
,
511
521
Swartzendruber
D. C.
,
Wertz
P. W.
,
Kitko
D. J.
,
Madison
K. C.
,
Downing
D. T.
(
1989
)
Molecular models of the intercellular lipid lamellae in mammalian stratum corneum.
J. Invest. Dermatol
92
,
251
257
Swartzendruber
D. C.
,
Wertz
P. W.
,
Madison
K. C.
,
Downing
D. T.
(
1987
)
Evidence that the corneocyte has a chemically bound lipid envelope.
J. Invest. Dermatol
88
,
709
713
Vernon
K. J.
,
Lane
A. T.
,
Wischerath
L. J.
,
Davis
J. M.
,
Menegus
M. A.
(
1990
)
Semi-permeable dressing and transepidermal water loss in the premature infant.
Pediatrics
86
,
357
362
Wertz
P. W.
,
Downing
D. T.
(
1987
)
Covalently bound-hydroxyacylsphingosine in the stratum corneum.
Biochim. Biophys. Acta
917
,
108
111
Wilson
D. R.
,
Maibach
H. I.
(
1980
)
Transepidermal water loss in vivo. Premature and term infants.
Biol. Neonate
37
,
180
185
Yuspa
S. H.
,
Kilkenny
A. E.
,
Steinert
P. M.
,
Roop
D. R.
(
1989
)
Expression of murine epidermal differentiation markers is tightly regulated by restricted extracellular calcium concentrations in vitro.
J. Cell Biol
109
,
1207
1217
This content is only available via PDF.