Geller S F, Lewis G P, Anderson D H, Fisher S K
Neuroscience Research Institute, University of California, Santa Barbara 93106-5060.
Invest Ophthalmol Vis Sci. 1995 Mar;36(3):737-44.
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.
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.
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.
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.
使用一种能识别增殖细胞中核特异性抗原Ki-67的抗体,研究视网膜内增殖作为对实验性视网膜脱离的一种反应。
在猫(1、3、7和28天)和兔(1、3和7天)中制造实验性视网膜脱离。处死动物,将眼睛固定并包埋在石蜡中。使用MIB-1抗体进行免疫组织化学处理组织学切片,以检测Ki-67蛋白。识别标记的细胞,并对增殖反应进行定量。
在正常猫视网膜中,每毫米视网膜约有0.05个细胞被标记。在视网膜脱离1、3、7或28天的猫视网膜中,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。与使用3H-胸腺嘧啶放射自显影术在视网膜切片中检测到的增殖细胞数量相比,MIB-1标记产生了约五倍的增加。兔视网膜脱离通过MIB-1免疫组织化学测量引发类似反应。
与仅标记S期细胞的3H-胸腺嘧啶不同,MIB-1抗体标记增殖细胞,无论其在细胞周期中的位置如何。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方法,并且是评估可能影响这种反应的药物作用的相对简单方法。