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目的分析嗅神经切断对小鼠嗅感觉神经元(olfactory receptor neurons,ORN)凋亡情况的影响,并探讨这一造模方法的可靠性。方法取2月龄C57小鼠33只,随机分组后,实验组小鼠行嗅神经切断术,通过辣根过氧化物酶(horseradis hperoxidase,HRP)顺行神经示踪验证造模成功与否。以脱氧核苷酸转移酶介导的生物素dUTP缺口末端标记技术(TdTmediat edde xyurid inetriphos phate biotinnic kendlabelling,TUNEL)观察术后8h、2d、3d和5d嗅上皮中ORN的凋亡情况,同时在蛋白和mRNA水平观察成熟ORN的特异性标记蛋白———嗅标记蛋白(olfactorymarkerprotein,OMP)在嗅上皮中的表达情况。结果嗅神经切断术后嗅球中无HRP标记。TUNEL和OMP阳性反应发生于ORN,嗅神经切断术后TUNEL阳性细胞数显著增多,并于术后第2天达到高峰,与此同时OMPmRNA的表达水平开始显著下降,并在术后第5天降至更低,嗅上皮的厚度也相应变薄。结论本实验所采取的造模方法可以较可靠地切断小鼠的嗅神经,并造成小鼠嗅上皮中ORN的同步凋亡。
Objective To analyze the effect of olfactory nerve transection on the apoptosis of olfactory receptor neurons (ORN) in mice and to explore the reliability of this method. Methods Thirty-two C57 mice of 2 months old were randomly divided into three groups. The mice in the experimental group were subjected to olfactory nerve transection, and the success of modeling was confirmed by horseradish hperoxidase (HRP). The apoptosis of ORN in the olfactory epithelium was observed at 8h, 2d, 3d and 5d after TdT mediated by deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) And mRNA levels were observed in mature ORN-specific marker protein --- olfactory marker protein (olfactorymarkerprotein, OMP) in the olfactory epithelium in the expression. Results There were no HRP markers in the olfactory bulb after olfactory nerve transection. TUNEL and OMP positive reaction occurred in ORN. The number of TUNEL-positive cells increased significantly after olfactory nerve transection, and reached its peak on the second day after operation. At the same time, the expression of OMP mRNA began to decline significantly, and decreased on the 5th day after operation To lower, the thickness of the sniffing epithelium also thinner accordingly. Conclusion The modeling method adopted in this study can cut the olfactory nerve of mice more reliably and cause synchronous apoptosis of ORN in the olfactory epithelium of mice.