Differential expression of laminin alpha chains during proliferative and differentiation stages in a model for skin morphogenesis.
The purpose of this study was to determine the mRNA and protein expression of laminin alpha chains at various stages of in vitro skin morphogenesis. Fibroblasts in mono-cultures express low levels of the mRNA of laminin alpha1,alpha2, alpha3 and alpha4 chains. When co-cultured with keratinocytes for 28 days, they expressed the mRNA for all these chains. Keratinocytes in monolayer expressed the laminin alpha3 chain mRNA and very low levels of the mRNA of the alpha1 and alpha2 chains, although, when recombined with fibroblasts they also expressed laminin alpha1and alpha2 mRNA, but not the laminin alpha4 mRNA. Immunocytochemistry of cells in co-culture showed that laminin alpha1, alpha3 and alpha5 chains were expressed in the epidermis, while the laminin alpha2, beta1, and gamma1 chains were noted in the dermis and at the epidermo-dermal interface. The laminin alpha1chain was first expressed during the proliferative stage (14-21 days) and the laminin alpha2 and alpha5 chains appeared later, during the differentiation stage (28-42 days). The above results suggest that epithelial-mesenchymal interactions are involved in the expression of laminin alpha chain mRNA during in vitro skin morphogenesis. In addition, there is distinct temporal and spatial expression of these chains during proliferative and differentiation stages, possibly reflecting different functions.
Fleischmajer
R
R
Department of Dermatology, Mount Sinai School of Medicine, Department of Dermatology, Box 1047, New York, NY 10029, USA. rf@doc.mssm.edu
Kuroda
K
K
Utani
A
A
Douglas MacDonald
E
E
Perlish
J S
JS
Arikawa-Hirasawa
E
E
Sekiguchi
K
K
Sanzen
N
N
Timpl
R
R
Yamada
Y
Y
D002454:Cell Differentiation; D002455:Cell Division; D002478:Cells, Cultured; D018920:Coculture Techniques; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism; D007797:Laminin / Q000235:genetics* / Q000378:metabolism; D009024:Morphogenesis; D012333:RNA, Messenger; D012867:Skin / Q000166:cytology / Q000378:metabolism*
C501369:LAMA4 protein, human; D007797:Laminin; D012333:RNA, Messenger; C102645:laminin alpha 2; C084098:laminin A; C095567:laminin alpha 3