TY - JOUR
T1 - Lack of nidogen-1 and -2 prevents basement membrane assembly in skin-organotypic coculture
AU - Nischt, Roswitha
AU - Schmidt, Cathrine
AU - Mirancea, Nicolae
AU - Baranowsky, Anke
AU - Mokkapati, Sharada
AU - Smyth, Neil
AU - Woenne, Eva C.
AU - Stark, Hans Jürgen
AU - Boukamp, Petra
AU - Breitkreutz, Dirk
N1 - Funding Information:
We thank all colleagues listed in “Materials and Methods” for providing antibodies and gratefully acknowledge the excellent technical assistance by Marion Reibetanz (Dermatology, Cologne) as well expert support by the photo department of the DKFZ. This work was supported by the Deutsche Forschungsgemeinschaft through the SFB 589 at the University of Cologne (R.N.) and through a project within the Priority Program “Basement Membranes” (C.S., E.C,W., D.B.), and further through an industrial grant (N.M., D.B.; Aventis Pharma Deutschland/Aventis-Sanofi).
PY - 2007/3
Y1 - 2007/3
N2 - Nidogens are considered as classical linkers joining laminin and collagen IV networks in basement membranes (BMs); however, recent genetic approaches have suggested that nidogens function in a tissue-specific and developmental context. Thus, in mice lacking both nidogen-1 and -2 heart and lung were severely affected, causing neonatal death. Furthermore, in various locations, extravasation of erythrocytes was observed implying microvascular defects. Mice expressing solely either isoform, had a functional BM, although nidogen-2 binds with lower affinity to the laminin γ1 chain. Having previously blocked BM formation by interfering with nidogen-1 binding to laminin in skin-organotypic cocultures, here we investigated the roles of nidogen-1 and -2 in this model. For that purpose, human HaCaT cells were grown in three-dimensional cocultures on collagen matrices containing murine fibroblasts of varying nidogen deficiency. As with our experiments blocking laminin-nidogen interaction, lack of both nidogens completely prevented BM deposition and ultrastructural assembly of BM and hemidesmosomes, although other BM proteins remained detectable at comparable levels with no signs of degradation. Supplementation by recombinant nidogen-1 or -2 restored these structures, as shown by immunofluorescence and electron microscopy, confirming that in this system nidogen-2 is equivalent to nidogen-1, and both can promote the development of a functional BM zone.
AB - Nidogens are considered as classical linkers joining laminin and collagen IV networks in basement membranes (BMs); however, recent genetic approaches have suggested that nidogens function in a tissue-specific and developmental context. Thus, in mice lacking both nidogen-1 and -2 heart and lung were severely affected, causing neonatal death. Furthermore, in various locations, extravasation of erythrocytes was observed implying microvascular defects. Mice expressing solely either isoform, had a functional BM, although nidogen-2 binds with lower affinity to the laminin γ1 chain. Having previously blocked BM formation by interfering with nidogen-1 binding to laminin in skin-organotypic cocultures, here we investigated the roles of nidogen-1 and -2 in this model. For that purpose, human HaCaT cells were grown in three-dimensional cocultures on collagen matrices containing murine fibroblasts of varying nidogen deficiency. As with our experiments blocking laminin-nidogen interaction, lack of both nidogens completely prevented BM deposition and ultrastructural assembly of BM and hemidesmosomes, although other BM proteins remained detectable at comparable levels with no signs of degradation. Supplementation by recombinant nidogen-1 or -2 restored these structures, as shown by immunofluorescence and electron microscopy, confirming that in this system nidogen-2 is equivalent to nidogen-1, and both can promote the development of a functional BM zone.
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U2 - 10.1038/sj.jid.5700562
DO - 10.1038/sj.jid.5700562
M3 - Article
C2 - 17008882
AN - SCOPUS:33847078062
SN - 0022-202X
VL - 127
SP - 545
EP - 554
JO - Journal of Investigative Dermatology
JF - Journal of Investigative Dermatology
IS - 3
ER -