Baghdassarian D, Toru-Delbauffe D, Gavaret J M, Pierre M
U.96 INSERM, Hôpital de Bicétre, Le Kremlin-Bicêtre, France.
Glia. 1993 Mar;7(3):193-202. doi: 10.1002/glia.440070302.
The present study was performed on primary cultures and subcultures of cerebellar astrocytes in order to investigate the effects of transforming growth factor-beta 1 (TGF beta 1) on proliferation, extracellular matrix (ECM) components, and cytoskeletal structures in relation to morphological changes. The expression and cellular distribution of the ECM components laminin and fibronectin and the cytoskeletal proteins glial fibrillary acidic protein (GFAP) and actin were investigated by immunoblotting, immunocytochemistry, and phalloidin staining. The proliferation of primary cultures was strongly inhibited by TGF beta 1. Treated cells became enlarged and spread onto the substratum. TGF beta 1 promoted the appearance of actin stress fibers and increased the cell actin content. It elicited a slight increase in GFAP expression and induced dispersion of thin filaments of GFAP. TGF beta 1 also stimulated the production of laminin and fibronectin and their incorporation into the ECM of primary cultures grown in medium with or without serum. Astrocytes grown in serum-containing medium for 1 day after subculturing responded strongly to TGF beta 1. Changes promoted by TGF beta 1 in cell shape, cytoskeleton, and ECM production of cultured astrocytes may have relevance for understanding the mechanisms of action of TGF beta 1 during brain development.
本研究在小脑星形胶质细胞的原代培养物和传代培养物上进行，以研究转化生长因子-β1（TGF-β1）对增殖、细胞外基质（ECM）成分和细胞骨架结构的影响及其与形态学变化的关系。通过免疫印迹、免疫细胞化学和鬼笔环肽染色研究了ECM成分层粘连蛋白和纤连蛋白以及细胞骨架蛋白胶质纤维酸性蛋白（GFAP）和肌动蛋白的表达及细胞分布。TGF-β1强烈抑制原代培养物的增殖。处理后的细胞变大并铺展在基质上。TGF-β1促进肌动蛋白应力纤维的出现并增加细胞肌动蛋白含量。它使GFAP表达略有增加，并诱导GFAP细丝分散。TGF-β1还刺激层粘连蛋白和纤连蛋白的产生，并将它们整合到在含血清或不含血清培养基中生长的原代培养物的ECM中。传代培养后在含血清培养基中培养1天的星形胶质细胞对TGF-β1反应强烈。TGF-β1促进培养的星形胶质细胞的细胞形状、细胞骨架和ECM产生的变化可能与理解TGF-β1在脑发育过程中的作用机制有关。
