Actin and Type I Collagen Propeptide Distribution in the Developing Chick Cornea
Actin and Type I Collagen Propeptide Distribution in the Developing Chick Cornea
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DOI：
10.1167/iovs.08-2554
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发表时间：
2009-04-01
影响因子：
4.4
通讯作者：
Ralphs, James R.
中科院分区：
文献类型：
--
作者：
Gealy, Claire;Hayes, Anthony J.;Ralphs, James R.
目的。测定鸡角膜基质发育过程中肌动蛋白丝的组织和I型前胶原的分布。对6 ~ 18日龄和产后18日龄的鸡胚角膜进行冷冻切片，用phalloidin荧光标记丝状肌动蛋白，用特异性单克隆抗体荧光标记I型前胶原的N端和c端前肽。用荧光显微镜和共聚焦显微镜观察组织切片。在胚胎各阶段均有明显的肌动蛋白丝束，呈正交排列。I型胶原前肽也存在，可见c -前肽为小灶，通常与肌动蛋白标签相关。间质基质中也检测到N-前肽，尤其是在鲍曼层中。直到胚胎第14天，角膜上皮中也有明显的肌动蛋白丝，并伴有胶原前肽标记。肌动蛋白丝束在基质中丰富，可能在发育中的鸡角膜的角化细胞中，并以正交方式排列，表明在该组织中细胞和基质组织中可能起作用。纤维束似乎与I型前胶原标记的病灶密切相关，提示肌动蛋白细胞骨架与胶原运输之间可能存在关联。I型胶原的N-前肽在细胞外基质中的存在和c -前肽的有限分布表明这些分子在分泌后的不同加工。n -前肽的持续存在意味着在发育中起作用，可能与胶原纤维直径和间距的控制有关。（中国眼科杂志，2009;50:1653-1658）DOI:10.1167/iovs.08-2554
PURPOSE. To determine the organization of actin filaments and distribution of type I procollagen during the development of the chick corneal stroma.METHODS. Embryonic chicken corneas of ages 6 to 18 days and 18 days posthatch were cryosectioned and fluorescently labeled for filamentous actin with phalloidin and for the N- and C-terminal propeptides of type I procollagen with specific monoclonal antibodies. Tissue sections were examined by fluorescence and confocal microscopy.RESULTS. Prominent actin filament bundles were present at all embryonic stages, arranged in orthogonal arrays. Type I collagen propeptides were also present, with the C-propeptide visible as small foci, often associated with the actin label. The N- propeptide was also detected in the stromal matrix, especially in Bowman's layer. Actin filaments were also prominent in the corneal epithelium, along with collagen propeptide labeling, up to embryonic day14.CONCLUSIONS. Actin filament bundles are abundant in the stroma, presumably in the keratocytes of the developing chick cornea, and are arranged in an orthogonal manner suggesting a possible role in cell and matrix organization in this tissue. Filament bundles appear to be closely associated with the foci of type I procollagen label, suggesting a possible association between the actin cytoskeleton and the trafficking of collagen. The presence of the N- propeptide of type I collagen in the extracellular matrix and the restricted distribution of the C-propeptide suggest differential processing of these molecules after secretion. The persistence of the N-propeptide implies a role in development, possibly in association with control of collagen fibril diameter and spacing. (Invest Ophthalmol Vis Sci. 2009; 50: 1653-1658) DOI:10.1167/iovs.08-2554