Coca-Prados M, Wax M B
Proc Natl Acad Sci U S A. 1986 Nov;83(22):8754-8. doi: 10.1073/pnas.83.22.8754.
Ciliary epithelial cells derived from human eye were successfully propagated through many generations after transformation with simian virus 40. The cell clone 8-SVHCE was isolated and characterized by immunoprecipitation and pharmacological studies that demonstrated the presence of several functional properties observed in the parent cells of this tissue. Immunoprecipitation revealed the presence of large tumor (T) antigen, and Southern blot analysis showed the incorporation of viral DNA into high molecular weight ciliary epithelial cell DNA. The presence of beta-adrenergic receptors was demonstrated by direct binding of a radiolabeled antagonist, [125I]iodopindolol, to membrane preparations of 8-SVHCE cells (Kd = 41.8 pM and Bmax = 67.1 fmol/mg of protein). Competition experiments with [125I]iodopindolol and selective drugs suggested that the receptors are of the beta 2-adrenergic subtype. Studies of catecholamine-stimulated cellular cAMP production and of isoproterenol-dependent protein phosphorylation of vimentin in 8-SVHCE indicated the functional conservation of beta-adrenergic receptor-mediated processes that are thought to be important in the regulation of aqueous humor production by the ciliary epithelium in vivo.
源自人眼的睫状上皮细胞在用猿猴病毒40转化后成功传代了许多代。分离出细胞克隆8-SVHCE，并通过免疫沉淀和药理学研究对其进行了表征，这些研究证明了在该组织的亲代细胞中观察到的几种功能特性的存在。免疫沉淀显示存在大T抗原，Southern印迹分析表明病毒DNA已整合到高分子量的睫状上皮细胞DNA中。通过放射性标记的拮抗剂[125I]碘吲哚洛尔与8-SVHCE细胞的膜制剂直接结合，证明了β-肾上腺素能受体的存在（Kd = 41.8 pM，Bmax = 67.1 fmol/mg蛋白质）。用[125I]碘吲哚洛尔和选择性药物进行的竞争实验表明，这些受体属于β2-肾上腺素能亚型。对8-SVHCE中儿茶酚胺刺激的细胞cAMP产生以及异异丙肾上腺素依赖性波形蛋白磷酸化的研究表明，β-肾上腺素能受体介导的过程在功能上具有保守性，这些过程被认为在体内睫状上皮调节房水生成中很重要。
