Deficiency in integrin-mediated transmembrane signaling and microfilament stress fiber formation by aging dermal fibroblasts from normal and Down's syndrome patients.
Deficiency in integrin-mediated transmembrane signaling and microfilament stress fiber formation by aging dermal fibroblasts from normal and Down's syndrome patients.
复制标题
正常人和唐氏综合症患者的老化真皮成纤维细胞缺乏整合素介导的跨膜信号传导和微丝应力纤维形成。
DOI：
10.1016/0014-4827(92)90022-z
复制
发表时间：
1992
影响因子：
3.7
通讯作者：
Culp,LA
中科院分区：
文献类型：
--
作者：
Flickinger,KS;Carter,WG;Culp,LA
以前的证据表明，在培养的人真皮成纤维细胞的短期放线菌酮治疗后，微丝应力纤维形成的缺陷，而细胞质扩散似乎完全正常，其他细胞骨架网络组织正常。这种缺陷适用于胶原基质（而不是纤连蛋白基质），并且对于体外老化的正常成纤维细胞和来自三种不同唐氏综合征患者的成纤维细胞在任何传代水平都是特异性的。为了鉴定衰老细胞中基质受体缺陷的机制，使用特异性单克隆抗体和两种互补的实验方法评价细胞中几种整联蛋白亚基的量和分布。流式细胞术分析表明，所有传代水平的所有这些细胞都具有大量的α3和β1整联蛋白亚基和少量的α5亚基，定向于纤连蛋白，其细胞表面可用性受放线菌酮处理的影响最小。相比之下，放线菌酮处理导致晚期传代乳头状和网状正常成纤维细胞以及所有三种唐氏病患者细胞中大部分α2亚基的表面可用性丧失，这些α 2亚基定向为胶原蛋白。细胞在补充抗坏血酸的培养基中的预先生长克服了应力纤维形成的缺陷，保留了大量的细胞表面可利用的α2亚基，可通过流式细胞术检测。当通过[35 S]甲硫氨酸放射性标记细胞的免疫沉淀评价整联蛋白亚基的量时，放线菌酮处理后任何细胞的α2亚基或任何其他亚基均未减少;然而，在衰老正常细胞和唐氏细胞中与β1复合的α2亚基少得多。因此，放线菌酮处理不会导致α2亚基数量的损失，而是导致其在细胞表面的掩蔽，并且不能通过质膜传递信号以影响应力纤维的形成。这种与衰老相关的整合素介导的信号传导缺陷现在可以用各种方法进行机械研究，以确定与整合素（顺式和/或反式作用分子）相互作用的细胞表面分子的性质，从而区分功能性受体和非功能性受体。
Previous evidence has shown a deficiency in microfilament stress fiber formation upon short-term cycloheximide treatment of cultured human dermal fibroblasts while cytoplasmic spreading appeared completely normal and other cytoskeletal networks organized normally. This deficiency applied to collagen substrata (not fibronectin substrata) and was specific forin vitro-aged normal fibroblasts and for fibroblasts from three different Down's syndrome patients at any passage level. To identify the mechanism(s) for matrix receptor deficiency in aging cells, cells were evaluated for amounts and distributions of several integrin subunits using specific monoclonal antibodies and two complementary experimental approaches. Flow cytometric analyses have shown that all these cells at all passage levels have large amounts of α3 and β1 integrin subunits and smaller amounts of the α5 subunit, directed to fibronectin, which are minimally affected in their cell surface availability by cycloheximide treatment. In contrast, cycloheximide treatment leads to the loss from surface availability of most of the α2 subunit, directed to collagen, in late-passage papillary and reticular normal fibroblasts and in all three Down's patient cells at all passages. Prior growth of cells in ascorbate-supplemented medium, which overcomes the deficiency in stress fiber formation, conserves the large amounts of cell surface-available α2 subunit detectable by flow cytometry. When amounts of integrin subunits were evaluated by immunoprecipitation of [35S]methionine-radiolabeled cells, there was no diminution of the α2 subunit or any other subunit for any cells upon cycloheximide treatment; however, there was much less α2 subunit complexed with β1 in aging normal and Down's cells. Therefore, cycloheximide treatment does not lead to loss in the amounts of the α2 subunit but rather to its masking at the cell surface and inability to transmit signals across the plasma membrane to effect stress fiber formation. This aging-related deficiency in integrin-mediated signaling can now be studied mechanistically with a variety of approaches to determine the nature of cell-surface molecules interacting with integrins (cis-and/ortrans-acting molecules) that discriminate functional from nonfunctional receptors.
登录
查看更多内容
细胞外基质组织调节成纤维细胞生长和生长因子反应性。
影响因子：
3.7
作者：
S. Nakagawa;Pamela Pawelek;F. Grinnell
通讯作者：
F. Grinnell
由神经节苷脂 GM1 和解整合素调节的背根神经元杂交克隆中的神经突生长。
影响因子：
3.7
作者：
Barletta,E;Bremer,EG;Culp,LA
通讯作者：
Culp,LA
巨噬细胞对层粘连蛋白的激活依赖性粘附涉及α6β1整合素的细胞骨架锚定和磷酸化。
DOI：
10.1083/jcb.110.6.2167
发表时间：
1990-06
期刊：
The Journal of cell biology
影响因子：
--
作者：
Shaw LM;Messier JM;Mercurio AM
通讯作者：
Mercurio AM
体外传代时纤连蛋白 mRNA 剪接的变化。
DOI：
--
发表时间：
1991
期刊：
Biochemical and Biophysical Research Communications - BBRC
影响因子：
--
作者：
E. M. Burke;D. Danner
通讯作者：
D. Danner
唐氏综合症患者的真皮成纤维细胞与正常衰老细胞一样，都存在放线菌酮诱导的胶原粘附反应缺陷。
影响因子：
3.7
作者：
Flickinger,KS;Culp,LA
通讯作者：
Culp,LA