Adult adenohypophysial cells express beta 1 integrins and prefer laminin during cell-substratum adhesion

Abstract

beta 1 Integrins are a family of structurally related heterodimeric cell surface receptors that are involved in adhesion to molecules in the extracellular matrix (ECM) such as laminin (LN), fibronectin (FN), and collagen. These receptors are expressed by many cell types and mediate a variety of processes such as cell-matrix and cell-to-cell adhesion, cell migration, growth, and differentiation. The purpose of these studies was to identify and partially characterize beta 1 integrins on adenohypophyseal cells and to begin to elucidate their functional importance. Adenohypophyses were removed from adult male rats, dispersed using 0.25% trypsin, rinsed, and resuspended in a 1:1 mixture of Dulbecco's modified Eagle's medium and F12 medium containing 10% fetal bovine serum and antibiotics. Ten million cells were allowed to attach to each of five plastic culture dishes overnight. The next day, the adenohypophyseal cells were surface-labeled with 125I. The labeled cells were lysed and centrifuged. The supernatant was immunoprecipitated using preimmune IgGs (100 micrograms/ml) and was then incubated with a polyclonal antibody against the rat beta 1 family of integrins or with a variety of immune IgGs directed against the alpha subunit of the receptor (anti alpha 1, anti alpha 2, anti alpha 3, and anti alpha 5 antibodies). The receptors were then immunoprecipitated by addition of protein A-Sepharose or IgG1 Sepharose. After washing, the immunoprecipitates were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis and autoradiography. Cultured adenohypophyseal cells expressed the beta 1 integrin subunit, which was associated with the alpha 1, alpha 2, alpha 3, and alpha 5 integrin subunits.(ABSTRACT TRUNCATED AT 250 WORDS)