Polarized expression of integrin receptors (α6β4, α 2β1, α3β1, and αvβ5) and their relationship with the cytoskeleton and basement membrane matrix in cultured human keratinocytes

Pier Carlo Marchisio, Sergio Bondanza, Ottavio Cremona, Ranieri Cancedda, Michele De Luca

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Abstract

In human keratinocytes cultured in conditions which allow differentiation and stratification and are suitable to reconstitute a fully functional epidermis, α6β4 and two members of the β1 integrin family (α2β1 and α3β1) were respectively polarized to the basal and lateral domains of the plasmamembrane both in growing colonies and in the reconstituted epidermis. Conversely, the αv integrin subunit, presumably in association with 185, was expressed at the basal surface in growing and migrating but not in stationary keratinocytes. The integrin α6β4: (a) was organized in typical patches which often showed a "leopard skin" pattern where spots corresponded to microfilament-free areas; (b) was not associated with focal contacts containing vinculin and talin but rather corresponded to relatively removed contact areas of the basal membrane as shown by interference reflection microscopy; and (c) was coherent to patches of laminin secreted and deposited underneath the ventral membrane of individual cells. The two β1 integrins (α2β1), both endowed with laminin receptor properties, were not associated with focal adhesions under experimental conditions allowing full epidermal maturation but matched the lateral position of vinculin (but not talin), cingulin, and desmoplakin, all makers of intercellular junctions. Often thin strips of laminin were observed in between the lateral aspects of individual basal keratinocytes. The integrin complex αvβ5 had a topography similar to that of talin- and vinculin-containing focal adhesions mostly in the peripheral cells of expanding keratinocyte colonies and in coincidence with fibronectin strands. The discrete topography of β1 and β4 integrins has a functional role in the maintenance of the state of aggregation of cultured keratinocytes since lateral aggregation was impaired by antibodies to β1 whereas antibodies to 184 prevented cell-matrix adhesion (De Luca, M., R. N. Tamura, S. Kajiji, S. Bondanza, P. Rossino, R. Cancedda, P. C. Marchisio, and V. Quaranta. Proc. Natl. Acad. Sci. USA. 87:6888-6892). Moreover, the surface polarization of integrins followed attachment and depended both on the presence of Ca2+ in the medium and on the integrity of the cytoskeleton. We conclude that our in vitro functional tests and structural data suggest a correlation between the pattern of integrin expression on defined plasmamembrane domains and the mechanism of epidermal assembly.

Original languageEnglish
Pages (from-to)761-773
Number of pages13
JournalJournal of Cell Biology
Volume112
Issue number4
Publication statusPublished - Feb 1991

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Cytoskeleton
Keratinocytes
Basement Membrane
Integrins
Talin
Vinculin
Focal Adhesions
Laminin
Epidermis
Desmoplakins
Panthera
Laminin Receptors
Interference Microscopy
Cell-Matrix Junctions
Intercellular Junctions
Antibodies
Actin Cytoskeleton
Fibronectins
Maintenance
Cell Membrane

ASJC Scopus subject areas

  • Cell Biology

Cite this

Polarized expression of integrin receptors (α6β4, α 2β1, α3β1, and αvβ5) and their relationship with the cytoskeleton and basement membrane matrix in cultured human keratinocytes. / Marchisio, Pier Carlo; Bondanza, Sergio; Cremona, Ottavio; Cancedda, Ranieri; De Luca, Michele.

In: Journal of Cell Biology, Vol. 112, No. 4, 02.1991, p. 761-773.

Research output: Contribution to journalArticle

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abstract = "In human keratinocytes cultured in conditions which allow differentiation and stratification and are suitable to reconstitute a fully functional epidermis, α6β4 and two members of the β1 integrin family (α2β1 and α3β1) were respectively polarized to the basal and lateral domains of the plasmamembrane both in growing colonies and in the reconstituted epidermis. Conversely, the αv integrin subunit, presumably in association with 185, was expressed at the basal surface in growing and migrating but not in stationary keratinocytes. The integrin α6β4: (a) was organized in typical patches which often showed a {"}leopard skin{"} pattern where spots corresponded to microfilament-free areas; (b) was not associated with focal contacts containing vinculin and talin but rather corresponded to relatively removed contact areas of the basal membrane as shown by interference reflection microscopy; and (c) was coherent to patches of laminin secreted and deposited underneath the ventral membrane of individual cells. The two β1 integrins (α2β1), both endowed with laminin receptor properties, were not associated with focal adhesions under experimental conditions allowing full epidermal maturation but matched the lateral position of vinculin (but not talin), cingulin, and desmoplakin, all makers of intercellular junctions. Often thin strips of laminin were observed in between the lateral aspects of individual basal keratinocytes. The integrin complex αvβ5 had a topography similar to that of talin- and vinculin-containing focal adhesions mostly in the peripheral cells of expanding keratinocyte colonies and in coincidence with fibronectin strands. The discrete topography of β1 and β4 integrins has a functional role in the maintenance of the state of aggregation of cultured keratinocytes since lateral aggregation was impaired by antibodies to β1 whereas antibodies to 184 prevented cell-matrix adhesion (De Luca, M., R. N. Tamura, S. Kajiji, S. Bondanza, P. Rossino, R. Cancedda, P. C. Marchisio, and V. Quaranta. Proc. Natl. Acad. Sci. USA. 87:6888-6892). Moreover, the surface polarization of integrins followed attachment and depended both on the presence of Ca2+ in the medium and on the integrity of the cytoskeleton. We conclude that our in vitro functional tests and structural data suggest a correlation between the pattern of integrin expression on defined plasmamembrane domains and the mechanism of epidermal assembly.",
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T1 - Polarized expression of integrin receptors (α6β4, α 2β1, α3β1, and αvβ5) and their relationship with the cytoskeleton and basement membrane matrix in cultured human keratinocytes

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AU - Bondanza, Sergio

AU - Cremona, Ottavio

AU - Cancedda, Ranieri

AU - De Luca, Michele

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N2 - In human keratinocytes cultured in conditions which allow differentiation and stratification and are suitable to reconstitute a fully functional epidermis, α6β4 and two members of the β1 integrin family (α2β1 and α3β1) were respectively polarized to the basal and lateral domains of the plasmamembrane both in growing colonies and in the reconstituted epidermis. Conversely, the αv integrin subunit, presumably in association with 185, was expressed at the basal surface in growing and migrating but not in stationary keratinocytes. The integrin α6β4: (a) was organized in typical patches which often showed a "leopard skin" pattern where spots corresponded to microfilament-free areas; (b) was not associated with focal contacts containing vinculin and talin but rather corresponded to relatively removed contact areas of the basal membrane as shown by interference reflection microscopy; and (c) was coherent to patches of laminin secreted and deposited underneath the ventral membrane of individual cells. The two β1 integrins (α2β1), both endowed with laminin receptor properties, were not associated with focal adhesions under experimental conditions allowing full epidermal maturation but matched the lateral position of vinculin (but not talin), cingulin, and desmoplakin, all makers of intercellular junctions. Often thin strips of laminin were observed in between the lateral aspects of individual basal keratinocytes. The integrin complex αvβ5 had a topography similar to that of talin- and vinculin-containing focal adhesions mostly in the peripheral cells of expanding keratinocyte colonies and in coincidence with fibronectin strands. The discrete topography of β1 and β4 integrins has a functional role in the maintenance of the state of aggregation of cultured keratinocytes since lateral aggregation was impaired by antibodies to β1 whereas antibodies to 184 prevented cell-matrix adhesion (De Luca, M., R. N. Tamura, S. Kajiji, S. Bondanza, P. Rossino, R. Cancedda, P. C. Marchisio, and V. Quaranta. Proc. Natl. Acad. Sci. USA. 87:6888-6892). Moreover, the surface polarization of integrins followed attachment and depended both on the presence of Ca2+ in the medium and on the integrity of the cytoskeleton. We conclude that our in vitro functional tests and structural data suggest a correlation between the pattern of integrin expression on defined plasmamembrane domains and the mechanism of epidermal assembly.

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