Investigation Of Odorant Eugenol Improving Olfactory Perception And Its Mechanism

ObjectiveOdorant or odor could have an effect on the brain through olfactory pathway.This study aims to confirm eugenol(EG) experience could improve olfactory perception in C57BL/6 mice,then to detect the change of olfactory receptor-mOR-EG gene expression in olfactory epithelium of mice which have been exposed to eugenol,and to investigate that odorant improves olfactory perception and its mechanism by detecting the changes of mOR-EG gene expression in olfactory sensory neurons which were cultured in EG enrichment in vitro.MethodsMice were divided into four groups from the PND0,including EG experience 6w group,6w blank control group,EG experience 10w group and 10w blank control group.Until the sixth week,10 mice were randomly chosen from EG 6w and 6w control group respectively and tested their behavior of odorant preference and odorant discrimination.So did EG 10w and 10w control in the tenth week.By observing the related quantiation of mOR-EG gene expression in olfactory epithelium of each group by Real-time PCR,we researched the possible relationship between the change of mOR-EG gene expression and the behavior of olfactory perception with EG experience.Then identified OSNs in vitro cell culture by characting OMP and TUJ-1 which were used to immunocytochemistry and RT-PCR technologies.Fianlly we processed with 30μm,100μm and 300μm EG enrichment in OSNs culture respectively to investigate the mechanism of EG experience improving olfactory perception furtherly by detecting the relative quantitation of mOR-EG gene expression of OSNs in each concentration of EG enrichment group and blank control group in vitro. Results1.Compared with 6w blank control group,the behavior data of EG 6w group demonstrated that EG experience improved olfactory discrimination in mice,and compared with 10w blank control group,EG 10w group also demonstrated that EG experience improve olfactory discrimination.2.Compared with 6w blank control group,the Real-time PCR data of EG 6w group demonstrate that EG experience elevated mOR-EG gene expression in olfactory epithelium,and compared with 10w blank control group,EG 10w group also demonstrated that EG experience elevated mOR-EG gene expression in olfactory epithelium.The mOR-EG expression of EG 6w was almost 9 times as much as EG 6w control,and EG 10w had the similar times than EG 10w control.3.To identify OSNs in vitro cell culture by characting OMP and TUJ-1 which were used to immunocytochemistry and RT-PCR technologies,OSNs manifested possitive expression of OMP and TUJ-1 proteins and genes.The EG enrichment experiment demonstrated that 30μM and 100μM EG was almost 2 and 4 times as much as control gene expression respectively,however,300μM EG had no significant difference from control.Conclusion1.EG experience could improve olfactory perception of EG in mice.2.EG experience could elevate the mOR-EG gene expression in olfactory epithelium in vivo.3.EG enrichment could elevate the mOR-EG gene expression in OSNs in vitro.