Horacek M J, Kawaguchi T, Terracio L
Department of Developmental Biology and Anatomy, University of South Carolina, School of Medicine, Columbia 29208.
In Vitro Cell Dev Biol Anim. 1994 Jan;30A(1):35-40. doi: 10.1007/BF02631416.
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)
β1整合素是一类结构相关的异二聚体细胞表面受体家族,参与细胞与细胞外基质(ECM)中分子的黏附,如层粘连蛋白(LN)、纤连蛋白(FN)和胶原蛋白。这些受体在多种细胞类型中表达,并介导多种过程,如细胞与基质及细胞间的黏附、细胞迁移、生长和分化。这些研究的目的是鉴定并部分表征腺垂体细胞上的β1整合素,并开始阐明它们的功能重要性。从成年雄性大鼠中取出腺垂体,用0.25%胰蛋白酶分散,冲洗后重悬于含有10%胎牛血清和抗生素的杜尔贝科改良伊格尔培养基与F12培养基的1:1混合物中。将1000万个细胞接种到五个塑料培养皿中,过夜贴壁。第二天,用125I对腺垂体细胞进行表面标记。标记的细胞经裂解和离心。上清液先用免疫前IgG(100微克/毫升)进行免疫沉淀,然后与抗大鼠β1整合素家族的多克隆抗体或针对受体α亚基的多种免疫IgG(抗α1、抗α2、抗α3和抗α5抗体)孵育。然后通过加入蛋白A-琼脂糖或IgG1琼脂糖对受体进行免疫沉淀。洗涤后,对免疫沉淀物进行十二烷基硫酸钠聚丙烯酰胺凝胶电泳和放射自显影。培养的腺垂体细胞表达β1整合素亚基,其与α1、α2、α3和α5整合素亚基相关。(摘要截断于250字)