Alpha 3 beta 1 integrin is moved into focal contacts in kidney mesangial cells.

The movement of integrins into focal adhesive structures accompanies cell attachment to extracellular matrix. The kinetics of incorporation of integrins into focal contacts was studied during attachment to matrix of mesangial cells of the kidney glomerulus. On collagen, fibronectin, laminin and vitronectin, the number and intensity of talin-focal contacts increased with time. Talin-containing focal contacts were present in mesangial cells within 2 h of plating and in control cells (HT1080 and Rugli) within 1 h. Integrin alpha-chains colocalized with talin, dependent on the matrix substrate. The attachment, spreading and organization of integrin into focal contacts was not affected when endogenous protein synthesis was suppressed with cycloheximide. In Rugli, alpha 1 beta 1 organized into focal contacts on collagen and laminin, while in HT1080 alpha 2 beta 1 organized on collagen type I, alpha 5 beta 1 on fibronectin, alpha 6 beta 1 on laminin, and alpha 3 beta 1 and alpha 4 beta 1 were diffusely distributed on all substrates. These distributions mirrored the usage and expression patterns previously established for integrins in these cells and was as predicted from the literature. In mesangial cells, however, alpha 3 beta 1 was also organized into prominent focal contact arrays on collagen, fibronectin, EHS and human placental laminins, but not on vitronectin, while alpha 6 beta 1 was not organized. Initial attachment and spreading of mesangial cells was absolutely dependent on divalent cations. Mg2+ and Mn2+ supported attachment on all substrates, while Ca2+ stimulated attachment on laminin (E8), fibronectin and vitronectin. The data suggest that the functional integrins on mesangial cells include alpha 1 beta 1 (on collagen and laminin) alpha 2 beta 1 (on collagen), alpha 5 beta 1 (on fibronectin) and alpha V beta 3 (on vitronectin). However, mesangial cells do not use alpha 6 beta 1 on laminin, and the data support a role for alpha 3 beta 1 as putative receptor for fibronectin, collagen and laminin.