嗅神经切断对小鼠嗅感觉神经元的影响  

Impact of olfactory nerve transection on the apoptosis of mice olfactory receptor neurons

作　　者：羡慕[1] 魏永祥[1] 韩德民[1] 范尔钟[2] 刘仲燕[2] 苗旭涛[1] 张聪[1] 张欣[1] 

机构地区：[1]首都医科大学附属北京同仁医院耳鼻咽喉头颈外科,100730 [2]北京市耳鼻咽喉科研究所

出　　处：《中华耳鼻咽喉头颈外科杂志》2005年第9期671-674,共4页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

基　　金：国家自然科学基金资助资助项目(30171004);北京市自然科学基金资助项目(7042012)

摘　　要：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 impact of olfactory nerve transection on the apoptosis of mice olfactory receptor neurons （ ORN）, and discuss the reliability of this experimental model. Methods After olfactory nerve transection of mice, anterograde horseradish peroxidase （ HRP ） tracing was performed to confirm the completion of nerve transection. On 8 h, 2 d, 3 d and 5 d after surgery, TdT mediated deoxyuridine triphosphate-biotin nick end labelling （ TUNEL ） was used to observe the apoptosis of ORN, while relative semi-quantitative RT-PCR and immunohistochemistry were used to reflect the expression of olfactory marker protein （ OMP, special marker of mature ORN ） in olfactory epithelium. Results No HRP label was observed in olfactory bulb after olfactory nerve transaction. Both TUNEL-possitive and OMP- positive cells were ORN. After the surgery, TUNEL-possitive cells increased remarkably and peaked on 2 d after the surgery. Meanwhile OMP mRNA in olfactory epithelium began to decrease markedly till 5 d after the surgery, and the olfactory epithelium got thinner accordingly. Conclusions This experimental model can be used reliably to sever mice olfactory nerve and manipulate simultaneous apoptosis of mice ORN.

关 键 词：嗅觉受体神经元 模型 动物 脱噬作用 C57小鼠 神经切断术 嗅感觉神经元 TUNEL阳性细胞 receptor mRNA水平 嗅标记蛋白 

分 类 号：R765[医药卫生—耳鼻咽喉科]