Lavail Matthew M, Nishikawa Shimpei, Duncan Jacque L, Yang Haidong, Matthes Michael T, Yasumura Douglas, Vollrath Douglas, Overbeek Paul A, Ash John D, Robinson Michael L
Beckman Vision Center, University of California, San Francisco, San Francisco, California 94143-0730, USA.
J Comp Neurol. 2008 Dec 20;511(6):724-35. doi: 10.1002/cne.21858.
Several neurotrophic factors (NTFs) are effective in protecting retinal photoreceptor cells from the damaging effects of constant light and slowing the rate of inherited photoreceptor degenerations. It is currently unclear whether, if continuously available, all NTFs can be protective for many or most retinal degenerations (RDs). We used transgenic mice that continuously overexpress the neurotrophin NT-3 from lens fibers under the control of the alphaA-crystallin promoter to test for neuroprotection in light-damage experiments and in four naturally occurring or transgenically induced RDs in mice. Lens-specific expression of NT-3 mRNA was demonstrated both by in situ hybridization in embryos and by reverse-transcriptase polymerase chain reaction (RT-PCR) in adult mice. Furthermore, NT-3 protein was found in abundance in the lens, ocular fluids, and retina by enzyme-linked immunosorbent assay (ELISA) and immunocytochemistry. Overexpression of NT-3 had no adverse effects on the structure or function of the retina for up to at least 14 months of age. Mice expressing the NT-3 transgene were protected from the damaging effects of constant light to a much greater degree than those receiving bolus injections of NT-3. When the NT-3 transgene was transferred into rd/rd, Rds/+, Q344ter mutant rhodopsin or Mertk knockout mice, overexpression of NT-3 had no protective effect on the RDs in these mice. Thus, specificity of the neuroprotective effect of NT-3 is clearly demonstrated, and different molecular mechanisms are inferred to mediate the protective effect in light-induced and inherited RDs.
几种神经营养因子(NTFs)可有效保护视网膜光感受器细胞免受持续光照的损伤,并减缓遗传性光感受器退化的速度。目前尚不清楚,如果持续存在,所有的神经营养因子是否都能对许多或大多数视网膜退化(RDs)起到保护作用。我们使用了在αA-晶状体蛋白启动子控制下从晶状体纤维持续过度表达神经营养蛋白NT-3的转基因小鼠,在光损伤实验以及小鼠的四种自然发生或转基因诱导的视网膜退化模型中测试神经保护作用。通过胚胎原位杂交和成年小鼠的逆转录聚合酶链反应(RT-PCR)均证实了NT-3 mRNA在晶状体中的特异性表达。此外,通过酶联免疫吸附测定(ELISA)和免疫细胞化学发现,NT-3蛋白在晶状体、眼液和视网膜中大量存在。NT-3的过度表达在至少14个月龄之前对视网膜的结构或功能没有不良影响。与接受单次注射NT-3的小鼠相比,表达NT-3转基因的小鼠在更大程度上免受持续光照的损伤。当将NT-3转基因转入rd/rd、Rds/+、Q344ter突变型视紫红质或Mertk基因敲除小鼠时,NT-3的过度表达对这些小鼠的视网膜退化没有保护作用。因此,NT-3神经保护作用的特异性得到了明确证明,并且推测不同的分子机制介导了光诱导和遗传性视网膜退化中的保护作用。