Institute of Clinical Sciences, Department of Ophthalmology, Lund University, Lund, Sweden.
Invest Ophthalmol Vis Sci. 2013 May 7;54(5):3272-80. doi: 10.1167/iovs.12-11363.
Progressive dynamic, relative quantitative changes were compared in glycans associated with retinal proteins of wild type (wt) and retinal degeneration 1 (rd1) mice during neonatal development and degeneration of retinae.
Proteins extracted from retinae of postnatal days 2 (PN2), PN7, and PN14 wt and rd1 mice were labeled with Cy3-fluorescent dye. Glycome of these proteins was quantified relatively by lectin microarray technique. Net fluorescence emitted by individual complexes formed between 45 lectins and Cy3-labeled proteins was measured by evanescent-field fluorescence-assisted microarray reader.
GlcNAcβ1-oligomer and high-mannose/Manα1-6Man were major glycans associated with the proteins of PN2, PN7, and PN14 wt and rd1 mice retinae. Gal/GalNAc/Man3-core-bi-/tri-antennary-complex, Sia2-3Galβ1-4GlcNAc, and high-mannose glycans were conjugated mainly to proteins from PN7 rd1 and PN14 wt retinae, respectively. With increasing neonatal age, mannosylated, GlcNAcβ, and sialylated (minor component) glycans were increased, and fucosylated GlcNAc/Galβ glycans were decreased significantly in wt retinal proteins. This trend was less evident in PN14 rd1 retinal proteins. Mouse retina was almost devoid of Siaα2-6 (except WGA bound Sia), Fucα1-2, and Gal/GalNAc-containing glycans. STL reacting GlcNAc oligomers were high in PN2 rd1 retinae.
Quantitative dynamic, relative variation in high-mannose and GlcNAc glycans, Siaα2-3Galβ1-4GlcNAc associated with proteins from PN2, PN7, and PN14 wt and rd1 mice retinae suggested that these glycans participate in retinal development and degeneration, and may be used as markers for retinal electrophysiologic integrity during transplantation/therapy studies; Siaα2-3Galβ1-4GlcNAc-specific Agrocybe cylindracea lectin and other lectins may be used to enrich/purify retinal ribbon synapse glycoproteins and other glycoproteins including rhodopsin. Further investigations are required.
比较野生型（wt）和视网膜变性 1 型（rd1）小鼠视网膜蛋白相关糖链在新生儿发育和视网膜变性过程中的渐进动态、相对定量变化。
用 Cy3 荧光染料标记出生后第 2 天（PN2）、PN7 和 PN14wt 和 rd1 小鼠视网膜中的蛋白质。通过凝集素微阵列技术相对定量糖组。通过瞬逝场荧光辅助微阵列读取器测量由 45 种凝集素与 Cy3 标记的蛋白质形成的单个复合物发出的净荧光。
GlcNAcβ1-寡聚物和高甘露糖/Manα1-6Man 是与 wt 和 rd1 小鼠视网膜蛋白相关的主要糖链。Gal/GalNAc/Man3 核心双/三触角复合、Sia2-3Galβ1-4GlcNAc 和高甘露糖糖链主要与 rd1 视网膜和 wtPN14 视网膜的蛋白质结合。随着新生儿年龄的增加，wt 视网膜蛋白中甘露糖化、GlcNAcβ 和唾液酸化（次要成分）糖增加，而 fucosylatedGlcNAc/Galβ 糖明显减少。这种趋势在 rd1 视网膜蛋白中不太明显。小鼠视网膜几乎不含 Siaα2-6（除 WGA 结合的 Sia）、Fucα1-2 和含有 Gal/GalNAc 的糖。STL 反应的 GlcNAc 寡聚物在 rd1 视网膜中含量较高。
wt 和 rd1 小鼠视网膜蛋白相关高甘露糖和 GlcNAc 糖、Siaα2-3Galβ1-4GlcNAc 的定量动态、相对变化表明，这些糖参与了视网膜的发育和变性，并且可能被用作视网膜电生理完整性的标志物在移植/治疗研究期间；Siaα2-3Galβ1-4GlcNAc 特异性金顶侧耳凝集素和其他凝集素可用于富集/纯化视网膜 ribbon 突触糖蛋白和其他糖蛋白，包括视紫红质。需要进一步研究。
