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.
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