Distribution of amyloid precursor protein and amyloid-beta in ocular hypertensive C57BL/6 mouse eyes.
Distribution of amyloid precursor protein and amyloid-beta in ocular hypertensive C57BL/6 mouse eyes.
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DOI：
10.3109/02713683.2010.494240
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发表时间：
2010-09
影响因子：
2
通讯作者：
Goldblum D
中科院分区：
文献类型：
--
作者：
Kipfer-Kauer A;McKinnon SJ;Frueh BE;Goldblum D
淀粉样前体蛋白（APP）和淀粉样β（Aβ）似乎参与青光眼视网膜神经节细胞（RGC）死亡的病理生理学。因此，我们测定了诱导慢性高眼压后C57 BL/6小鼠视网膜中APP和Aβ的分布。通过将高渗盐水注射到巩膜外静脉中，在三个月大的C57 BL/6小鼠的一只眼睛中诱导高眼压。记录到眼内压（IOP）升高6周后，用4%多聚甲醛固定视网膜，并用抗体进行免疫组织化学处理，包括Aβ 40 C末端的多克隆抗体（Novartis 17-40/23）和APP胞外域的多克隆抗体（Novartis 474）。以盲法对高眼压和对照视网膜中APP和Aβ免疫标记的分布和半定量表达进行分级并进行比较。APP和Aβ免疫反应在高眼压视网膜的软脑膜/硬脑膜、视神经（ON）和RGC层中发现，而对侧对照眼的APP和Aβ免疫反应仅在软脑膜/硬脑膜中检测到。高眼压眼和对照眼Aβ免疫标记的比较在ON和RGC层中是显著的（p < 0.05），而APP染色的比较没有发现显著差异。在高眼压视网膜中观察到高Aβ和APP水平，可能是由于IOP升高时APP剪接异常所致。
Amyloid precursor protein (APP) and amyloid-beta (Aβ) appear to participate in the pathophysiology of retinal ganglion cell (RGC) death in glaucoma. We, therefore, determined the distribution of APP and Aβ in the retinas of C57BL/6 mice after induction of chronic ocular hypertension. Ocular hypertension was induced in one eye of three-month-old C57BL/6 mice by injection of hypertonic saline into episcleral veins. After 6 weeks of documented elevated intraocular pressure (IOP), retinas were fixed with 4% paraformaldehyde and processed for immunohistochemistry with antibodies including a polyclonal antibody to the C-terminus of Aβ 40 (Novartis 17-40/23) and a polyclonal antibody to the APP ectodomain (Novartis 474). Distribution and semiquantitative expression of APP and Aβ immunolabeling in ocular hypertensive and control retinas were graded in a masked fashion and compared. APP and Aβ immunoreactivity was found in the pia/dura, optic nerve (ON), and RGC layer of ocular hypertensive retinas, whereas APP and Aβ immunoreactivity in the contralateral control eyes was detected only in the pia/dura. Comparison of ocular hypertensive and control eyes for Aβ immunolabeling was significant in the ON and RGC layer (p < 0.05) whereas no significant difference was found when compared for APP staining. High Aβ and APP levels were seen in ocular hypertensive retinas, probably due to abnormal APP-splicing in the presence of elevated IOP.