Phenotypic changes and loss of N-CAM-mediated adhesion in transformed embryonic chicken retinal cells.
Phenotypic changes and loss of N-CAM-mediated adhesion in transformed embryonic chicken retinal cells.
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DOI：
10.1083/jcb.99.6.1944
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发表时间：
1984-12
期刊：
影响因子：
--
通讯作者：
Edelman GM
中科院分区：
文献类型：
--
作者：
Brackenbury R;Greenberg ME;Edelman GM
Rous肉瘤病毒（RSV）转化6日龄鸡胚视网膜细胞，发现引起显着的变化，在几个细胞特性，包括存活，运动，和分化状态。细胞粘附性的改变进行了分析，通过特异性抗体的钙非依赖性神经细胞粘附分子，N-CAM。在RSV转化的细胞中，存在于细胞表面的N-CAM的量相对于正常细胞显著降低，如通过免疫荧光染色、特异性免疫沉淀和免疫印迹实验所评估的。这种减少反映在体外测量的N-CAM介导的炎症的显著减少中。一个不同的，钙依赖性，粘附系统也存在于神经元上的RSV转化没有检测到改变，与以前的研究对正常神经元相比，这种粘附系统被检测到没有蛋白酶治疗。在培养中，转化细胞形成的集落比正常视网膜细胞更少，更不紧凑。通过延时电影摄影术观察RSV转化的视网膜细胞证实了细胞增殖的减少，并且还揭示了转化的细胞比它们的正常对应物具有更高的运动性。此外，RSV转化似乎改变了培养的视网膜细胞的分化。免疫荧光染色研究表明，与成熟神经元相反，转化的神经视网膜细胞表达34，000-mol-wt酪氨酸激酶底物，并减少单克隆抗体A2 B5识别的神经元特异性神经节苷脂的量。这些特征是未转化的神经胶质细胞所共有的。在双重免疫荧光染色实验中，许多细胞在病毒感染后不久同时表达N-CAM和pp 60 src，这意味着N-CAM阳性的神经上皮细胞被RSV转化。此外，使用荧光激活细胞分选仪选择的高度纯化的N-CAM阳性神经视网膜细胞群体被RSV以与未分级群体相当的速率快速且广泛地转化。这些结果表明，转化细胞主要来源于RSV感染的神经上皮细胞，而不是来自原代培养物中存在的视网膜神经胶质细胞的小群体。研究结果表明，重新考虑肿瘤的可能起源分类的形态学标准，来自神经胶质细胞和提高的可能性，pp 60 src的正常同系物可能发挥作用的承诺，神经上皮细胞的神经元或神经胶质细胞分化途径。
Transformation of 6-d-old embryonic chicken retinal cells by Rous sarcoma virus (RSV) was found to cause significant changes in several cellular properties including adhesiveness, motility, and state of differentiation. The alterations in cell adhesivity were analyzed by means of specific antibodies to the calcium-independent neural cell adhesion molecule, N-CAM. In the RSV-transformed cells the amount of N- CAM present at the cell surface was significantly decreased relative to normal cells, as assessed by immunofluorescent staining, specific immunoprecipitation, and immunoblotting experiments. This decrease was reflected in a marked reduction in N-CAM-mediated adhesiveness measured in vitro. A different, calcium-dependent, adhesive system also present on neurons was not detectably altered by RSV transformation and, in contrast with previous studies on normal neurons, this adhesive system was detected without treatment by proteases. In culture, the transformed cells formed fewer and less compact colonies than the normal retinal cells. Observation of the RSV-transformed retinal cells by time-lapse cinematography confirmed the reduction in adhesiveness and also revealed that the transformed cells were more highly motile than their normal counterparts. In addition, RSV transformation appeared to alter the differentiation of the cultured retinal cells. Immunofluorescent staining studies indicated that in contrast to mature neurons, transformed neural retinal cells expressed the 34,000-mol-wt tyrosine kinase substrate and reduced amounts of a neuron-specific ganglioside recognized by monoclonal antibody A2B5. These characteristics are shared by untransformed glial cells. In double immunofluorescent staining experiments, many cells expressed both N-CAM and pp60src shortly after viral infection, which implies that the N-CAM- positive neuroepithelial cells were transformed by RSV. In addition, a highly purified population of N-CAM-positive neural retinal cells, selected using a fluorescence-activated cell sorter, was rapidly and extensively transformed by RSV at rates comparable to those of the unfractionated population. These results established that the transformed cells were largely derived from RSV-infected neuroepithelial cells rather than from a small population of retinal glial cells present in the primary culture. The findings suggest reconsideration of the possible origin of tumors classified by morphological criteria as derived from glia and raise the possibility that the normal homologue of pp60src may play a role in the commitment of neuroepithelial cells to neuronal or glial differentiation pathways.
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DOI：
10.1073/pnas.79.22.7036
发表时间：
1982-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
EDELMAN, GM;CHUONG, CM
通讯作者：
CHUONG, CM
影响因子：
2.9
作者：
CHAMBERLAIN, JP
通讯作者：
CHAMBERLAIN, JP
影响因子：
7.8
作者：
GRUMET, M;EDELMAN, GM
通讯作者：
EDELMAN, GM
DOI：
10.1038/newbio231147a0
发表时间：
1971-01-01
期刊：
NATURE-NEW BIOLOGY
影响因子：
--
作者：
EDWARDS, JG;CAMPBELL, JA;WILLIAMS, JF
通讯作者：
WILLIAMS, JF
DOI：
10.1073/pnas.81.3.969
发表时间：
1984-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
GREENBERG, ME;BRACKENBURY, R;EDELMAN, GM
通讯作者：
EDELMAN, GM