Institute of Reconstructive Neurobiology, Medical Faculty & University Hospital Bonn, University of Bonn, Bonn, Germany.
Experimental Immunology of the Eye, Department of Ophthalmology, University Hospital Cologne, Cologne, Germany.
Glia. 2024 Dec;72(12):2295-2312. doi: 10.1002/glia.24613. Epub 2024 Sep 3.
Sialylation plays an important role in self-recognition, as well as keeping the complement and innate immune systems in check. It is unclear whether the reduced sialylation seen during aging and in mice heterozygous for the null mutant of UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (Gne+/-), an essential enzyme for sialic acid biosynthesis, contributes to retinal inflammation and degeneration. We found a reduction of polysialic acid and trisialic acid expression in several retinal layers in Gne+/- mice at 9 months of age compared to Gne+/+ wildtype (WT) mice, which was associated with a higher microglial expression of the lysosomal marker CD68. Furthermore, the total number of rod bipolar cells was reduced in 12 months old Gne+/- mice in comparison to WT mice, demonstrating loss of these retinal interneurons. Transcriptome analysis showed up-regulation of complement, inflammation, and apoptosis-related pathways in the retinas of Gne+/- mice. Particularly, increased gene transcript levels of the complement factors C3 and C4 and the pro-inflammatory cytokine Il-1β were observed by semi-quantitative real-time polymerase chain reaction (sqRT-PCR) in 9 months old Gne+/- mice compared to WT mice. The increased expression of CD68, loss of rod bipolar cells, and increased gene transcription of complement factor C4, were all prevented after crossing Gne+/- mice with complement factor C3-deficient animals. In conclusion, our data show that retinal hyposialylation in 9 and 12 months old Gne+/- mice was associated with complement-related inflammation and lysosomal microglia response, as well as rod bipolar cells loss, which was absent after genetic deletion of complement factor C3.
唾液酸化在自我识别中起着重要作用,同时还能控制补体和先天免疫系统。目前尚不清楚衰老过程中以及在 UDP-N-乙酰氨基葡萄糖 2-差向异构酶/ N-乙酰甘露糖胺激酶(Gne+/-)杂合子缺失突变的小鼠中观察到的唾液酸化减少是否导致视网膜炎症和变性。我们发现与 Gne+/+野生型(WT)相比,9 月龄 Gne+/-小鼠的几个视网膜层中的多唾液酸和三唾液酸表达减少,这与溶酶体标记物 CD68 的小胶质细胞表达增加有关。此外,与 WT 相比,12 月龄 Gne+/-小鼠的杆状双极细胞总数减少,表明这些视网膜中间神经元丢失。转录组分析显示 Gne+/-小鼠的视网膜中补体、炎症和细胞凋亡相关途径上调。特别是,通过半定量实时聚合酶链反应(sqRT-PCR)观察到 Gne+/-小鼠中补体因子 C3 和 C4 以及促炎细胞因子 Il-1β的基因转录水平升高。与 WT 相比,9 月龄 Gne+/-小鼠中 CD68 的表达增加、杆状双极细胞丢失和补体因子 C4 的基因转录增加,在用缺乏补体因子 C3 的动物与 Gne+/- 杂交后均得到预防。总之,我们的数据表明,9 和 12 月龄 Gne+/- 小鼠的视网膜低唾液酸化与补体相关炎症和溶酶体小胶质细胞反应以及杆状双极细胞丢失有关,而在用补体因子 C3 基因缺失消除后则不存在。