目的探讨小鼠晶状体细胞增殖、细胞周期蛋白(cyclin)D1和突触素(SYN)的表达情况。方法各日龄小鼠共150只,HE染色和4’6-二脒基-2-苯基吲哚(DAPI)染色,在光镜下观察小鼠晶状体的一般结构;用抗细胞增殖核抗原(PCNA)免疫荧光法以及5’-溴脱氧尿嘧啶核苷(BrdU)技术标记晶状体增殖细胞;用免疫荧标记标价法检测细胞周期蛋白cyclinD1和SYN在晶状体的表达。结果孕8d(E8)左右,晶状体由最初的晶状体板分化而来,是由单层柱状上皮细胞围成的空泡,空泡再进一步分化,被原始晶状体纤维所替代,逐渐形成晶状体;BrdU和PCNA的阳性细胞主要分布在晶状体上皮细胞的前囊中央区,但出生10d(P10)以后BrdU阳性细胞已不在晶状体上皮细胞分布,而PCNA持续表达。Cyclin D1阳性细胞在胚胎期主要在晶状体上皮细胞的赤道部表达,出生早期在晶状体上皮细胞的前囊中央区,出生5d(P5)以后在赤道前区分布,赤道部有少量阳性细胞;在晶状体发育过程中,SYN可以在晶状体基质中和赤道部胞体中发现。结论细胞增殖对晶状体的发育和修复起着重要的作用;cyclin D1可能参与调控赤道部细胞的消失,因此保持晶状体的透明度;SYN在晶状体的表达说明了它对晶状体的透明有着重要的作用,通过调控钙离子内流人胞体和基质中。
Objective To study cell proliferation and the expression of cell cycle protein (eyclin D1 ) and synaptophysin (SYN) in mouse lens. Methods A total 150 mice were used in the study. HE staining and DAPI staining were performed out to observe the general structure of the mouse lens. Proliferating cell nuclar antigen (PCNA) immunofluorescent staining and 5-bromodeoxyuridine (BrdU) assay were used to mark the proliferative cells in lens. The expression of cyclin D1 protein and synaptophysin in the lens were investigated with cyclin D1 and synaptophysin immunofluorescent staining. Results At about E8, the lens were evolved from the lens placode with single columnar epithelium. Then, the hollow lens bulb developed further and was filled with numerous fibroblasts, which gradually formed lens. BrdU and PCNA-positive ceils were mainly distributed in the epithelium of anterior capsule. However, PIO afterward, BrdU positive cells disappeared, while PCNA continued to express in the anterior epithelial ceils. Cyclin D1- positive cells were mainly distributed in the epithelium of anterior capsule and the equator of the lens in the embryonic days and early postnatal days. After P5, cyclinDl-positive cells were mainly located in the pre-equatorial area, and only few cyclin D1 positive cells were found in the equator. Synaptophysin protein expressed in the matrix and some cell bodies of equator during the development. Conclusion The cell proliferation plays an important role during the lens development and lens repair. Cell cycle proteins, cyclin D1 is probably involved in regulating the cells' disappear in the equator, therefore the lens ' s transparency can be kept. Synaptophysin ' s expression in lens has important function in lens ' s transparency, through its regulation to calcium flow in cell bodies and matrix of lens.