Assessment of rapid morphological changes associated with elevated cAMP levels in human orbital fibroblasts

被引:34
作者
Reddy, L
Wang, HS
Keese, CR
Giaever, I
Smith, TJ
机构
[1] Albany Med Coll, Div Mol & Cellular Med A175, Dept Med, Albany, NY 12208 USA
[2] Albany Med Coll, Dept Biochem & Mol Biol, Albany, NY 12208 USA
[3] Rensselaer Polytech Inst, Sch Sci, Troy, NY 12180 USA
[4] Appl BioPhys Inc, Troy, NY 12180 USA
[5] Samuel S Stratton Vet Affairs Med Ctr, Albany, NY 12208 USA
关键词
Graves' disease; thyroid associated ophthalmopathy; electric cell-substrate impedance sensing; cytokine action;
D O I
10.1006/excr.1998.4273
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Orbital fibroblasts exhibit a phenotype distinct from that of other types of fibroblasts. Addition of prostaglandin E-2 (PGE(2)) to culture medium elicits a dramatic change in orbital fibroblast morphology, That response is mediated through the generation of cAMP, Orbital fibroblasts can generate high levels of PGE(2) through induction by proinflammatory cytokines of prostaglandin endoperoxide H synthase-2 (PGHS-2). Here we compare the influence on fibroblast morphology of exogenous PGE(2), forskolin, and 8-br-cAMP to that mediated through PGHS-2 induction by a lymphocyte-derived cytokine, Within a few hours, orbital fibroblasts treated with any of these test compounds appear under phase-contrast microscopy to exhibit a stellate morphology, When these changes were assessed quantitatively by electric cell-substrate impedance sensing (ECIS), it became evident that 8-br-cAMP, forskolin, and PGE(2) initiated shape changes within 30 min of addition to the culture medium, while effects of the cytokine were first evident after approximately 3.5 h, Dermal fibroblasts failed to respond to any of these compounds with regard to changes in cellular morphology. Analysis of micromotion, manifested as small impedance fluctuations, revealed that orbital fibroblasts treated with 8-br-cAMP exhibit less motion than did untreated cells, These results suggest that orbital fibroblast shape can be altered by several compounds known to alter intracellular cAMP levels. They demonstrate the utility of ECIS in the assessment of very rapid and dynamic cellular events associated with changes in cell morphology, (C) 1998 Academic Press.
引用
收藏
页码:360 / 367
页数:8
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