CPG15在发育期正常和单眼形觉剥夺大鼠视皮层表达的研究

Study of CPG15 expression in the visual cortex of normal and monocular form deprived development rats

目的 研究CPG15 mRNA和蛋白在发育期正常和单眼形觉剥夺（MD）大鼠视皮层中的表达,探讨其与视觉发育的相关性.方法 实验研究.80只健康Wistar大鼠分为正常组和MD组,根据发育的不同时限,正常组观察时间点分别为生后7、14、21、28、45 d,MD组分别为生后21、28和45 d,每个时间点10只大鼠.MD组大鼠于出生后14 d行右侧眼睑缝合制作单眼形觉剥夺模型.所有大鼠在同等环境下饲养.采用SYBRgreen Ⅰ荧光定量RT-PCR和免疫组织化学方法检测CPG15mRNA和CPG15阳性细胞数及蛋白表达的IOD值在发育期正常和MD大鼠视皮层中的表达变化.正常组和MD组间数据行单因素方差分析,同一时间点的两组数据采用成组设计的t检验进行统计学分析.结果 生后7、14、21、28、45 d时,正常组大鼠视皮层中CPG15 mRNA相对表达量分别为0.152±0.011、0.234±0.052、0.527±0.047、0.846±0.067、0.445±0.220,MD组大鼠则在生后21、28、45 d时相对表达量分别为0.412±0.015、0.501±0.019、0.381±0.030,组间比较差异均有统计学意义（F=177.52,36.93;P＜0.01）.两组大鼠视皮层中CPG15 阳性细胞数及蛋白表达的IOD值组间比较也均有统计学意义（F=26.001,12.207,76.914,58.397;P＜0.01）.CPG15 mRNA和蛋白表达的IOD值均在视觉发育关键期的中期P28和MD28达高峰.MD组与正常组大鼠视皮层中CPG15mRNA和CPG15 阳性细胞数及蛋白表达的IOD值在生后21、28、45 d比较差异均有统计学意义,MD组其表达量均明显减少.结论 发育期正常和MD大鼠视皮层中CPG15 mRNA和CPG15阳性细胞数及蛋白表达的IOD值随视觉发育呈阶段性变化,其表达与视觉经验和年龄因素相关,在视觉发育关键期呈高水平表达,单眼形觉剥夺明显影响视皮层中CPG15 mRNA和蛋白的表达,推测CPG15参与视皮层神经元的发育和成熟,它可能是参与视觉发育可塑性变化的分子基础之一.

Objective To explore the expression of CPG15 mRNA and protein the visual cortex of normal and monocular form deprived development rats, To explore the associatiion CPG15 with visual development. Methods It was an experimental research. One hundred healthy Wistar rats were divided into 10 groups： the time-point for normal group is postnatal 7,14,21,28,45 days. The time-point for MD group is postnatal 21,28,45 days, 10 in every group. MD groups were sutured right eyelid at postnatal 14 days,and to make the model of monocular form deprived. Rats were housed in alike environment. To detect the expression of Neuritin mRNA and immuno-positive neurons protein in the visual cortex of normal and monocular form deprived rats by SYBRgreen Ⅰ real-time fluorescent polymerase chain reaction and visual cortex CPG15 mRNA and immuno-positive neurons and the IOD of protein, significant deviation existed among different ages with one-factor analysis of variance（ P 〈0.05 ）, and reached its climax in P28group obviously lower than the normal group in postnatal 21,28,45 days ,there was statistically different in that ages（P 〈0.05）. Conclusion The expression of CPG15 mRNA and immuno-positive neurons and the IOD of protein in the development normal and MD rats show that the stage change by development vision. Its expression correlated with visual experience and age factor, and there was a high expression in critical period,and obviously influence by monocular form deprived. So we supposed that CPG15 may affect development and maturation in the visual cortical neuron. It may be the one of molecular basis to participated in visual developmental plasticity.