Use of the MIB-1 antibody for detecting proliferating cells in the retina.
Use of the MIB-1 antibody for detecting proliferating cells in the retina.
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DOI：
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发表时间：
1995-03
影响因子：
4.4
通讯作者：
S. Geller;G. Lewis;Don H. Anderson;S. Fisher
中科院分区：
文献类型：
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作者：
S. Geller;G. Lewis;Don H. Anderson;S. Fisher
目的利用一种能识别增殖细胞核特异性抗原Ki-67的抗体研究视网膜内增殖对实验性视网膜脱离的反应。方法用猫（1、3、7、28 d）和兔（1、3、7 d）制作实验性视网膜脱离模型。处死动物，将眼睛固定并包埋在石蜡中。使用MIB-1抗体对组织切片进行免疫组织化学处理，以检测Ki-67蛋白。鉴定标记的细胞，并定量增殖反应。结果在正常猫视网膜中，每毫米视网膜约有0.05个细胞被标记。在猫视网膜脱离1、3、7或28天，由MIB-1标记的细胞数分别为每毫米视网膜0.06、5.03、1.38和0.23个细胞。MIB-1标记产生的增殖细胞的数量增加了约5倍，在视网膜切片中检测到使用3 H-胸苷放射自显影。兔视网膜的脱离也引起类似的反应，如通过MIB-1免疫组织化学所测量的。结论与仅标记S期细胞的3 H-胸苷相反，MIB-1抗体标记增殖细胞，而不管它们在细胞周期中的位置。因此，MIB-1标记是评估视网膜和中枢神经系统其他部位细胞增殖的更准确的方法，并且是评估可能影响这种反应的药物的作用的相对简单的方法。
PURPOSE To study intraretinal proliferation as a response to experimental retinal detachment using an antibody that recognizes the nuclear specific antigen Ki-67 in proliferating cells. METHODS Experimental retinal detachments were produced in cats (1, 3, 7, and 28 days) and rabbits (1, 3, and 7 days). The animals were killed and the eyes were fixed and embedded in paraffin. Histologic sections were processed for immunohistochemistry using the MIB-1 antibody to detect the Ki-67 protein. Labeled cells were identified, and the proliferative response was quantified. RESULTS In normal cat retina, approximately 0.05 cells per millimeter of retina are labeled. In cat retina detached for 1, 3, 7, or 28 days, the number of cells labeled by MIB-1 is 0.06, 5.03, 1.38, and 0.23 cells per millimeter of retina, respectively. MIB-1 labeling yields an approximate fivefold increase over the number of proliferating cells detected in retinal sections using 3H-thymidine autoradiography. Detachment of the rabbit retina elicits a similar response as measured by MIB-1 immunohistochemistry. CONCLUSIONS In contrast to 3H-thymidine, which labels cells in S-phase only, the MIB-1 antibody labels proliferating cells regardless of their location within the cell cycle. MIB-1 labeling, therefore, is a more accurate means of evaluating cellular proliferation in the retina and elsewhere in the central nervous system, and it is a relatively simple way of evaluating the effects of agents that may affect this response.