Effect of FGFs on adult bovine Muller cells: proliferation, binding and internalization.
Effect of FGFs on adult bovine Muller cells: proliferation, binding and internalization.
复制标题
FGF 对成年牛 Muller 细胞的影响：增殖、结合和内化。
DOI：
10.3109/08977199109000260
复制
发表时间：
1991
期刊：
影响因子：
1.8
通讯作者：
Y. Courtois
中科院分区：
文献类型：
--
作者：
F. Mascarelli;J. Tassin;Y. Courtois
本文报道了一种从成年牛组织中培养视网膜Muller细胞的新方法。这些神经胶质细胞的鉴定是基于特定的Muller细胞标志物的免疫细胞化学分析。第4 ~ 9代培养细胞的S-100蛋白、碳酸酐酶（CAA）、谷氨酰胺合成酶（GS）、α-cristallin（α-C）和多克隆胶质细胞酸性蛋白（GFAP）抗体均呈阳性，而单克隆GFAP抗体和视网膜Muller细胞均呈阴性。研究酸性成纤维细胞生长因子（aFGF）、碱性成纤维细胞生长因子（bFGF）和上皮生长因子（EGF）对Muller细胞增殖的影响，发现bFGF是最有效的有丝分裂原（EC 50 = 14 pM）。结合数据揭示了aFGF和bFGF的两类结合位点的存在：（1）高亲和力结合位点（aFGF和bFGF的Kd分别为14 pM和27 pM）;（2）低亲和力结合位点（aFGF和bFGF的Kd分别为3.2nM和0.6nM，结合位点的数目具有很大的可变性）。此外，交联实验揭示了高分子量FGF受体（110-140 kDa）的存在。aFGF或bFGF与Muller细胞结合后，aFGF和bFGF-细胞表面受体迅速下调，半衰期为35-50 min。4 ℃时，与Muller细胞受体结合的125 I-bFGF未发生内化和降解。然而，在37 ℃时，由于bFGF受体的下调，受体结合的125 I-bFGF迅速减少。125 I-bFGF在Muller细胞内出现。2 h后，125 I-bFGF开始降解，6 h后可分辨出16 kDa、8 kDa和5.5kDa的三个片段。碱性成纤维细胞生长因子的降解似乎发生在溶酶体室，因为它被抑制氯喹，溶酶体蛋白酶的抑制剂; aFGF的内化和降解遵循与碱性成纤维细胞生长因子相同的动力学与7 kDa和5 kDa的片段的外观。这些结果表明，Muller细胞可能是视网膜其他细胞中所含aFGF和bFGF的靶点。aFGF可通过磷酸化依赖性机制从视杆外节释放，并且Muller细胞的顶端延伸与感光细胞相关，这一事实表明这些因子可能是参与胶质细胞和神经元之间通讯的介质。
A new method for culturing retinal Muller cells from adult bovine tissue is described. The identification of these glial cells was based on immunocytochemical analysis of specific Muller cell markers. Cultured cells from fourth to ninth passage showed positive labelling for S 100 protein, carbonic anydrase (CAA), glutamine synthetase (GS), alpha cristallin (alpha C) and polyclonal glial fibrillary acidic protein (GFAP) antibody, but were negative for both monoclonal GFAP antibody and also for Muller cells in the retina. Investigation of the effect of acidic fibroblast growth factor (aFGF), basic fibroblast growth factor (bFGF), and epithelial growth factor (EGF) on the proliferation of the Muller cells revealed that bFGF was the most potent mitogen (EC50 = 14 pM). Binding data revealed the presence of two classes of binding sites for aFGF and bFGF: (1) a high affinity binding site (Kd of 14 pM and 27 pM for aFGF and bFGF respectively); (2) a low affinity binding site (Kd of 3.2 nM and 0.6 nM for aFGF and bFGF respectively with great variability in the number of binding sites). In addition, the cross-linking experiments revealed the presence of high molecular weight FGF receptors (110-140 kDa). After aFGF or bFGF binding to Muller cells, aFGF and bFGF-cell surface receptors were rapidly downregulated with a half-life for disappearance of 35-50 min. Internalization and degradation of 125I-bFGF bound to the Muller cell receptors did not occur at 4 degrees C. At 37 degrees C, however, there was a rapid decrease in receptor-bound 125I-bFGF due to the downregulation of bFGF receptors. Concomitantly 125I-bFGF appeared inside the Muller cells. After 2 h, 125I-bFGF began to be degraded and after 6 h three fragments of 16 kDa, 8 kDa and 5.5 kDa were discernible. Degradation of bFGF appeared to occur in the lysosomal compartment since it was inhibited by chloroquine, an inhibitor of lysosomal proteases; aFGF internalization and degradation followed the same kinetics as bFGF with the appearance of 7 kDa and 5 kDa fragments. These results suggest that Muller cells may be the target for aFGF and bFGF contained in other cells of the retina. The fact that aFGF could be released from rod outer segment by a phosphorylation-dependent mechanism, and that apical prolongation of the Muller cells is connected with the photoreceptor cells suggest that these factors may be the mediators involved in the communication between glial cells and neurons.
登录
查看更多内容
视网膜母细胞瘤中的神经胶质细胞成分。
影响因子：
3.4
作者：
Craft,JL;Sang,DN;Dryja,TP;Brockhurst,RJ;Robinson,NL;Albert,DM
通讯作者：
Albert,DM
DOI：
10.1016/s0021-9258(18)68179-7
发表时间：
1988-10
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
J. Feige;A. Baird
通讯作者：
J. Feige;A. Baird
小仓鼠肾细胞成纤维细胞生长因子受体的鉴定和部分表征。
DOI：
--
发表时间：
1985
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Neufeld,G;Gospodarowicz,D
通讯作者：
Gospodarowicz,D
内皮细胞对肝素结合生长因子-I 的内化和降解。
DOI：
10.1016/s0006-291x(88)80459-5
发表时间：
1988
影响因子：
3.1
作者：
Friesel,R;Maciag,T
通讯作者：
Maciag,T
DOI：
--
发表时间：
1986
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Huang,JS;Huang,SS;Kuo,MD
通讯作者：
Kuo,MD