Fibronectin extra domain A (FN-EDA) causes glaucomatous trabecular meshwork, retina, and optic nerve damage in mice.
Fibronectin extra domain A (FN-EDA) causes glaucomatous trabecular meshwork, retina, and optic nerve damage in mice.
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DOI：
10.1186/s13578-022-00800-y
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发表时间：
2022-05-26
影响因子：
7.5
通讯作者：
McDowell, Colleen M.
中科院分区：
文献类型：
--
作者：
Mavlyutov, Timur A.;Myrah, Justin J.;Chauhan, Anil K.;Liu, Yang;McDowell, Colleen M.
眼内压（IOP）升高是原发性开角型青光眼发生和发展的主要危险因素，并且是由于小梁网（TM）损伤所致。在这里，我们调查的作用，内源性Toll样受体4（TLR 4）配体，FN-EDA，在青光眼的发展利用转基因小鼠品系（B6.EDA+/+），组成型表达只有FN含有EDA亚型。对来自C57 BL 6/J（野生型）、B6.EDA+/+（组成型活性EDA）、B6.EDA-/-（EDA无效）小鼠的眼睛进行电子显微镜处理，并收集TM和施累姆氏管（SC）从前部到后部的整个长度的连续图像，并将其拼接成单个图像。通过ImageJ分析定量ECM积累、基底膜长度以及巨空泡的大小和数量。采用RNAscope原位杂交和免疫组织化学方法检测TM和ONH细胞中Tlr 4和Iba 1的表达。使用回弹眼压计测量IOP，通过PPD染色评估ON损伤，并在RBPMS标记的视网膜平坦封片中定量RGC损失。 超微结构分析显示，与C57 BL/6 J小鼠相比，B6.EDA+/+小鼠的TM在TM梁之间具有显著增加的ECM积累，具有很少的空隙（p < 0.05）。与C57 BL/6 J相比，B6.EDA+/+小鼠的SC基底膜更厚且更连续。在EDA敲除小鼠的TM中未检测到显著的结构差异。与C57 BL/6 J眼相比，B6.EDA+/+小鼠的TM中Tlr 4和Iba 1表达增加（p < 0.05）。与C57 BL/6 J眼睛相比，B6.EDA+/+小鼠的IOP显著更高（p < 0.001），并且在1岁时检测到显著的ON损伤（p < 0.001）和RGC损失（p < 0.05）。Tlr 4 mRNA在小鼠ONH细胞中表达，并且存在于神经节细胞轴突、小胶质细胞和星形胶质细胞中。与C57 BL/6 J对照眼相比，在B6.EDA+/+小鼠的ONH中Iba-1阳性小胶质细胞占据的面积显著增加（p < 0.01）。B6.EDA+/+小鼠具有TM中ECM积累增加、IOP升高、ONH中促炎性变化增强、RGC损失和ONH损伤。这些数据表明，B6.EDA+/+小鼠重演了脑昏迷损伤的许多方面。在线版本包含补充材料，可通过10.1186/s13578-022-00800-y获得。
Elevated intraocular pressure (IOP) is a major risk factor for the development and progression of primary open angle glaucoma and is due to trabecular meshwork (TM) damage. Here, we investigate the role of an endogenous Toll-like receptor 4 (TLR4) ligand, FN-EDA, in the development of glaucoma utilizing a transgenic mouse strain (B6.EDA+/+) that constitutively expresses only FN containing the EDA isoform. Eyes from C57BL6/J (wild-type), B6.EDA+/+ (constitutively active EDA), B6.EDA-/- (EDA null) mice were processed for electron microscopy and consecutive images of the entire length of the TM and Schlemm’s canal (SC) from anterior to posterior were collected and montaged into a single image. ECM accumulation, basement membrane length, and size and number of giant vacuoles were quantified by ImageJ analysis. Tlr4 and Iba1 expression in the TM and ONH cells was conducted using RNAscope in situ hybridization and immunohistochemistry protocols. IOP was measured using a rebound tonometer, ON damage assessed by PPD stain, and RGC loss quantified in RBPMS labeled retina flat mounts. Ultrastructure analyses show the TM of B6.EDA+/+ mice have significantly increased accumulation of ECM between TM beams with few empty spaces compared to C57BL/6 J mice (p < 0.05). SC basement membrane is thicker and more continuous in B6.EDA+/+ mice compared to C57BL/6 J. No significant structural differences are detected in the TM of EDA null mice. Tlr4 and Iba1 expression is increased in the TM of B6.EDA+/+ mice compared to C57BL/6 J eyes (p < 0.05). IOP is significantly higher in B6.EDA+/+ mice compared to C57BL/6 J eyes (p < 0.001), and significant ON damage (p < 0.001) and RGC loss (p < 0.05) detected at 1 year of age. Tlr4 mRNA is expressed in mouse ONH cells, and is present in ganglion cell axons, microglia, and astrocytes. There is a significant increase in the area occupied by Iba-1 positive microglia cells in the ONH of B6.EDA+/+ mice compared to C57BL/6 J control eyes (p < 0.01). B6.EDA+/+ mice have increased ECM accumulation in the TM, elevated IOP, enhanced proinflammatory changes in the ONH, loss of RGCs, and ONH damage. These data suggest B6.EDA+/+ mice recapitulate many aspects of glaucomatous damage. The online version contains supplementary material available at 10.1186/s13578-022-00800-y.
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影响因子：
15.9
作者：
Howell, Gareth R.;Soto, Ileana;John, Simon W. M.
通讯作者：
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影响因子：
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作者：
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作者：
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