Centripetal movement of corneal epithelial cells in the normal adult mouse
Centripetal movement of corneal epithelial cells in the normal adult mouse
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DOI：
10.1167/iovs.02-0705
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发表时间：
2003-02-01
影响因子：
4.4
通讯作者：
Zhao, J
中科院分区：
文献类型：
--
作者：
Nagasaki, T;Zhao, J
目的。目的通过组织学和活体显微镜观察正常成年小鼠角膜上皮细胞的自然运动。方法利用绿色荧光蛋白(GFP)基因工程构建的转基因小鼠株系，对角膜上皮细胞进行显影。在组织学方面，在荧光显微镜下，对面部完整的角膜上皮GFP进行成像，并在冰冻切片上进行成像。F，或体内研究，麻醉的小鼠被放在一个定制的观察平台上，在荧光显微镜下。每周数字化记录角膜上皮细胞荧光2~3次，并根据时间推移序列测定细胞运动速度。结果：出生后1周左右，角膜上皮细胞中GFP的表达几乎无处不在，此后逐渐变得零星，导致GFP阳性细胞的镶嵌模式，最亮的细胞出现在上皮的基底层和基底层。高绿色荧光蛋白细胞和低绿色荧光蛋白细胞都向中央角膜形成放射状条纹，经常在中央显示漩涡图案。在存活的小鼠体内跟踪了长达7周的几组高GFP CFF，时间推移序列分析表明它们向心移动的平均速度为26um/d。结论成年GFP小鼠的角膜上皮细胞呈现出一种适合于研究正常角膜细胞运动的GFP表达模式，这些小鼠角膜基底层或上基底层的上皮细胞向心移动的平均速度为26um/d。
PURPOSE. To study the natural movement of corneal epithelial cells in the normal adult mouse with histology and in vivo microscopy.METHODS. A transgenic mouse line that was engineered to ubiquitously, express green fluorescent protein (GFP) was used to visualize corneal epithelial cells. For histology, epithelial GFP was imaged in a wholemounted cornea en face, and also in frozen cross-sections, under a fluorescence microscope. F, or in vivo studies, the anesthetized mouse was placed on a custom-made observation platform under a fluorescence microscope. Epithelial fluorescence was digitally recorded two to three times a week, and a rate of cell movement was determined from the time-lapse sequences.RESULTS. The GFP expression in the corneal epithelium was nearly ubiquitous up to about I week after birth, and thereafter it gradually became sporadic, resulting in a mosaic pattern of GFP positive cells, with the brightest cells present in the basal and suprabasal layer of the epithelium. Both high- and low GFP-cells formed radial streaks toward the central cornea, frequently displaying vortex patterns at the center. Clusters of several high-GFP ceffs were tracked in living mice for up to 7 weeks, and an analysis of time-lapse sequences revealed that they moved centripetally at an average rate of 26 mum/d.CONCLUSIONS. Corneal epithelium of adult GFP mice exhibits a pattern of GFP expression that is suitable for studying cell movement in the normal cornea, Epithelial cells at the basal or suprabasal layers move centripetally in these mice at an average rate of 26 mum/d.