Glaucoma Research Laboratory, Wilmer Eye Institute, 600 North Wolfe Street, Johns Hopkins University School of Medicine, Baltimore, MD 21287-9205, USA.
Exp Eye Res. 2011 Apr;92(4):299-305. doi: 10.1016/j.exer.2011.01.006. Epub 2011 Jan 25.
To determine if the absence of c-Jun N-terminal kinase 3 (JNK3) in the mouse retina would reduce retinal ganglion cell (RGC) loss in mice with experimental glaucoma. C57BL/6 mice underwent experimental intraocular pressure (IOP) elevation with a bead/viscoelastic injection into one eye. One-half of the mice were Jnk3 homozygous knockouts (KO) and were compared to wild type (WT) mice. IOP was measured under anesthesia with the TonoLab, axial length was measured post-mortem with calipers after inflation to 15mmHg, and RGC layer counts were performed on retinal whole mount images stained with DAPI, imaged by confocal microscopy, and counted by masked observers in an image analysis system. Axon counts were performed in optic nerve cross-sections by semi-automated image analysis. Both WT and Jnk3(-/-) mice had mean elevations of IOP of more than 50% after bead injection. Both groups underwent the expected axial globe elongation due to chronic IOP elevation. The absence of JNK3 in KO retina was demonstrated by Western blots. RGC layer neuron counts showed modest loss in both WT and Jnk3(-/-) animals; local differences by retinal eccentricity were detected, in each case indicating greater loss in KO animals than in WT. The baseline number of RGC layer cells in KO animals was 10% higher than in WT, but the number of optic nerve axons was identical in KO and WT controls. A slightly greater loss of RGC in Jnk3(-/-) mice compared to controls was detected in experimental mouse glaucoma by RGC layer counting and there was no protective effect shown in axon counts. Counts of RGC layer cells and optic nerve axons indicate that Jnk3(-/-) mice have an increased number of amacrine cells compared to WT controls.
为了确定在实验性青光眼小鼠中缺失 c-Jun N-末端激酶 3（JNK3）是否会减少视网膜神经节细胞（RGC）的损失。C57BL/6 小鼠通过在一只眼内注射珠/粘弹性物质来进行实验性眼压（IOP）升高。一半的小鼠是 Jnk3 纯合敲除（KO）小鼠，并与野生型（WT）小鼠进行比较。在麻醉下使用 TonoLab 测量 IOP，在充气至 15mmHg 后用卡尺测量死后的眼轴长度，并用 DAPI 染色的视网膜全层图像进行 RGC 层计数，通过共聚焦显微镜成像，并在图像分析系统中由蒙面观察者进行计数。视神经横切片中的轴突计数通过半自动图像分析进行。珠注射后，WT 和 Jnk3（-/-）小鼠的平均 IOP 升高均超过 50%。两组均因慢性 IOP 升高而发生预期的眼球伸长。通过 Western blot 证明 KO 视网膜中缺乏 JNK3。RGC 层神经元计数显示 WT 和 Jnk3（-/-）动物均有适度损失；在每个情况下，通过视网膜偏心度检测到局部差异，表明 KO 动物的损失大于 WT。KO 动物的 RGC 层细胞的基线数量比 WT 高 10%，但 KO 和 WT 对照组的视神经轴突数量相同。与对照组相比，实验性小鼠青光眼的 RGC 层计数检测到 Jnk3（-/-）小鼠的 RGC 丢失略多，但在轴突计数中未显示出保护作用。RGC 层细胞和视神经轴突的计数表明，与 WT 对照相比，Jnk3（-/-）小鼠的无长突细胞数量增加。
