Hedgehog Olfactory Bulb C-fos, Nos Expression And No Content In Seasonal Differences

Hibernation is an adaptive dormancy to resist negative living conditions such as low temperatures in winter. Nowadays heterothermic animals's hibernation and hibernation mechanism have been paid more and more attentions, but neural physiological regulatory mechanism during hibernation is still not very clear. This thesis adopts hedgehog (Erinaceus europaeus) as experimental materials, aims at providing the theory base for neural physiological regulatory mechanism of hibernation.The histological structure was studied by means of HE stain in hedgehog's olfactory bulb, with a view of enriching the comparative histology data to provide certain basis for anatomical atla's establishment of hedgehog's olfactory bulb histological structure. To investigate the relativity of c-Fos, olfaction and hibernation, immunohistochemistry is adopted to test the seasonal differences of c-Fos expression in each level of hedgehog's olfactory bulb. In order to investigate the relation between c-Fos, olfaction and hibernation, NADPH-diaphorase histochemistry was used to study the distribution and quantity of NOS-positive neurons in hedgehog's olfactory bulb in different seasons. Nitrate reductase enzymatic is used to detect the seasonal differences of NO purity in hedgehog's olfactory. The photos were loaded into Motic Images Advanced 3.2 for measure and statistics of expression ratio were analyzed in GraphPad Prism4 and Microsoft Excel software was used to plot. The results were showed as follows:1. Hedgehog's olfactory bulb looks like a concentric circle on a transverse section and exhibits a typical laminar organization. From the outside to inside, the levels in sequence of hedgehog's olfactory bulb substance were olfactory nerve layer, olfactory glomerular layer, external plexiform layer, mitral cell layer, internal plexiform layer, granule cell layer and ependymal layer. The thickness of each level have a great difference on different transverse section and cell density varied greatly in different level.2. Strong expression of c-Fos protein is tested in each level among adult hedgehog's olfactory bulb, while active shade is not noticed when performing negative control. The result showss that seasonal differences are obviously displayed. The general trend is:expression ratio of c-Fos protein is lowest in winter while highest in fall. It indicates that fall is the most active period of hedgehog's olfactory neurons while hedgehog's olfactory neurons activity significantly reduced in winter. Strong expression of c-Fos protein in hedgehog's olfactory indicates that c-Fos protein may play a significant role in transferring the hedgehog's olfactory message. Significant seasonal differences of c-Fos expression show that there is correlation between hedgehog's olfactory bulb activity and its hibernation. 3. Seasonal differences of nitric oxide synthase (NOS) expression in Hedgehog's olfactory bulb:NADPH-diaphorase histochemistry was well demonstrated the expression of NOS-positive neurons in hedgehog's olfactory bulb. The result shows that NOS positive neurons were widely distributed in hedgehog's olfactory bulb. Compared with non-hibernation, NOS-positive neurons is deeper-colored and high expression in hibernation:As we know, hibernation is the extension of sleep and olfactory bulb plays a role in controlling animal cycle rhythm. The results may indicate that:in order to maintain hibernating state, olfactory bulb may need produce much more NO.NOS is an essential enzyme in the process of NO synthesis, so it need more NOS to synthesize more NO to keep in hibernating state.4. Seasonal differences of amount of nitric oxide in hedgehog's olfactory bulb:compared with pre-hibernation, hibernation and period after hibernating, we found that the amount of nitric oxide in hedgehog's olfactory is lowest in winter and highest in fall, but statistic data shows that there is no significant difference in these three periods. We guess:mammalian anabolic reaction decrease during hibernation, amount of L-Arg becomes the constraint in the process of NO synthesis. Meanwhile, mammalian respiration slows down during hibernation, amount of molecular oxygen should be another limiting factor. But there is no significant difference in these three periods indicates that NO shows great activity in hibernation.