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刺猬嗅球冬眠期与非冬眠期c-Fos 表达的差异

龙娣,郭炳冉,高玲,江乐,高燕,卢少俊   

  1. 曲阜师范大学生命科学学院
  • 出版日期:2011-08-31 发布日期:2011-03-02

The differences of c-Fos expression in Hedgehog (Erinaceus europaeus)olfactory bulb during hibernation and non-hibernation

  • Online:2011-08-31 Published:2011-03-02

摘要: 利用免疫组化法检测c-Fos 蛋白在不同季节刺猬嗅球各层次的表达差异,探讨c-Fos、嗅觉、冬眠三者的关系。分别选取春、夏、秋、冬四个季节各6 只野生健康刺猬,固定剥离嗅球,石蜡切片,免疫组化显色,拍片,载入Motic Images Advanced 3.2 软件,测量四个季节刺猬嗅球各层次c-Fos 的表达率,将结果载入GraphPadPrism4 软件分析,Microsoft Excel 作图。结果表明:c-Fos 蛋白在成年刺猬嗅球各层均有不同程度的表达,阴性对照不着色,且表现出明显的季节性差异。1)与秋季相比,冬眠期c-Fos 蛋白在刺猬嗅球各层次的表达均有极显著的降低(P <0.01);2)与夏季相比,冬眠期c-Fos 蛋白在外网丛层、僧帽细胞层、颗粒细胞层的表达有极显著降低(P < 0. 01),在嗅神经层、嗅小球层、室管膜层的表达也有显著降低(P < 0. 05);3)与冬眠期相比,春季c-Fos 蛋白在嗅小球层、僧帽细胞层、颗粒细胞层的表达有极显著的升高(P <0. 01),在嗅神经层、外网丛层、室管膜层的表达却没有显著变化(P ﹥ 0.05);4)嗅神经层c-Fos 的表达在春季显著低于秋季,夏季与秋季没有显著差异。颗粒细胞层夏季显著低于秋季(P < 0.05)。秋季c-Fos 在其余各层次的表达与春季、夏季相比都有极显著的提高(P <0.01)。结论:秋季刺猬嗅球神经元最活跃,嗅觉最灵敏,冬眠期刺猬嗅球活跃性大大降低,嗅觉系统最迟钝。c-Fos 在刺猬嗅球中的强表达表明其在嗅觉信息的传递中可能发挥一定作用,c-Fos 表达率的显著季节性差异揭示了刺猬嗅球的活跃性与其冬眠具有一定的相关性。

关键词: 刺猬, 嗅球, c-Fos, 免疫组化, 冬眠

Abstract: To investigate the relationships of c-Fos,olfaction and hibernation,immunohistochemistry is used to test the seasonal differences in c-Fos expression in each level of hedgehog olfactory bulb. The experimental process was as follows: six wild hedgehogs of each season in good shape were selected,and then anaesthetized and perfused,the olfactory bulbs were fixed and peeled,and finally make paraffin-embedded sections were cut. The cut sections were imaged and measured by immunohistochemistry and photos were taken. The photos were loaded into Motic Images Advanced 3. 2,to measure the expression ratio of c-Fos in each level of hedgehog olfactory bulb of each season. Statistics of expression ratio were analyzed in GraphPad Prism4. Microsoft Excel software was used to plot the data. Strong expression of c-Fos protein was found in each level among adult hedgehog olfactory bulb,while staining is not found in negative controls. The results showed that seasonal differences were obviously displayed. The general trend was:expression ratio of c-Fos protein was lowest in winter while highest in fall. 1)The expression of c-Fos in each level of hedgehog olfactory bulb in winter were significantly lower than that of in summer and fall (P <0.01);2)The expression of c-Fos in the external plexiform layer,mitral cell layer
and granule cell layer in winter were significantly lower than that in summer (P <0.01). In the olfactory nerve layer,the olfactory glomerular layer and the ependymal layer,the expression of c-Fos also was lower than that in summer (P <0.05);3)Compared with winter,c-fos was significantly up-regulated at first in spring in the olfactory glomerular layer, the mitral cell layer and the granule cell layer (P <0. 01),while there were no differences among the olfactory nerve layer, the external plexiform layer,or the ependymal layer (P ﹥ 0.05);4)Compared with fall,the expression of c-Fos in the olfactory nerve layer in spring increased significantly while no differences when compared expression in summer. In the rest level of hedgehog’s olfactory bulb,the expression of c-Fos in fall is significantly higher than that in spring and summer. Strong expression of c-Fos protein in hedgehog’s olfactory bulb indicates that c-Fos protein may play an significant
role in transferring hedgehog’s olfactory messages. Significant seasonal differences of c-Fos expression show that there is correlation between hedgehog olfactory bulb activity and its hibernation.

Key words: c-Fos, Hedgehog, Hibernation, Immunohistochemistry, Olfactory bulb