Microglia are highly plastic cells that participate in inflammatory and injury responses within the CNS and that can migrate extensively after activation. Because astrocytes and their extracellular matrix (ECM) form a large part of the CNS parenchyma, we undertook to study the adhesive interactions between microglia and these substrates in vitro. In contrast to oligodendrocyte precursor cells, microglia formed only weak interactions with astrocytes and their ECM. On specific ECM substrates the microglia adhered strongly to fibronectin, vitronectin, and plastic but only weakly to laminin. Microglial adhesion to laminin was increased significantly by the proinflammatory cytokines TNF, IFN-alpha, and IFN-gamma but was decreased by TGF-beta1, with the TGF-beta1 effect being dominant over the other cytokines. Fluorescence-activated cell sorting (FACS) analysis and immunoprecipitation showed that microglia constitutively express the alpha6beta1 integrin, a well characterized laminin receptor, and that alpha6beta1 expression levels did not change after cytokine treatment. Function-blocking studies showed that microglial adhesion to laminin is mediated entirely by the alpha6beta1 integrin, strongly suggesting that the cytokine regulation of adhesion to laminin is mediated by changes in the activation state of alpha6beta1. Analysis of signaling pathways revealed that activation of alpha6beta1 is mediated by a PKC-dependent mechanism. In light of the evidence that laminin expression is upregulated after CNS injury, the findings suggest that cytokine regulation of microglial adhesion to laminin may play a fundamental role in determining the extent of microglial infiltration into and retention at the site of injury.