摘要
主要嗅觉表皮(main olfactory epithelium,MOE)是哺乳动物感知气味分子的主要嗅觉器官。在MOE组织内,大多数嗅觉神经元通过c AMP信号传导通路感知气味信息。作为嗅觉c AMP信号通路的主要成员之一,腺苷酸环化酶3(adenylyl cyclase 3,ac3)基因敲除小鼠嗅觉探测功能丧失。除c AMP信号传导通路外,MOE内AC3相关因子AC2和AC4,以及肌醇1,4,5-三磷酸(inositol 1,4,5-trisphosphate,IP3)信号通路和Sonic Hedgehog(Shh)信号通路均有表达。然而,敲除ac3是否会对ac2和ac4以及IP3和Shh信号通路成员产生影响,尚不清楚。本文以AC3缺失(AC3-/-)及其野生型小鼠(AC3+/+)MOE为材料,采用实时荧光定量PCR(qRT-PCR)和免疫荧光组织化学方法,发现AC3缺失后,MOE内的ac2和ac4,以及IP3信号通路中的IP3受体ip3r1及钙调蛋白calm1和calm2表达水平均明显降低。Shh信号通路中的受体patched(ptch)与smoothened(smo)、以及核转录因子gli1与gli2的表达也受到了影响。总之,AC3基因缺失不但导致小鼠MOE组织中c AMP信号通路受损,同时AC3相关因子,IP3信号通路和Shh信号通路的传导也受到抑制。本文对于阐明AC3基因敲除小鼠嗅觉丧失的原因及其嗅觉探测机制具有重要启示作用。
The main olfactory epithelium(MOE) is the primary olfactory organ for odorant perception inmammals.Within the MOE,c AMP signal transduction pathway is the main pathway for the detection of odor molecules.As one of the key components of c AMP signaling,adenylyl cyclase 3(AC3) deletion leads to the defects in olfactory detection in mice.In addition to c AMP signaling,the AC3 isoforms,adenylyl cyclase 2(AC2) and adenylyl cyclase 4(AC4) associated with the c AMP signaling pathway,the inositol 1,4,5,-trisphosphate(IP3) and the sonic hedgehog(Shh) signaling pathway components are also expressed in the MOE.However,whether the expression level of AC2,AC4,and the cascade members of IP3 and Shh signaling,will be altered in the MOE after AC3 deletion in mice remains to be determined.In the present study,by real-time fluorescent quantitative PCR(qRT-PCR) and immunofluorescence staining approaches,with MOE from AC3-/-and AC3+/+mice,we demonstrated that the relative expression of ac2 and ac4 was down-regulated in the MOE of AC3-/-mice,compared with AC3+/+controls.In addition,expression level of inositol 1,4,5-trisphosphate receptor 1(ip3r1),calmodulin(CaM) genes calm1,calm2,patched(ptch),smoothened(smo),the nuclear transcription factors gli1 and gli2,components associated with IP3 and Shh signaling pathways,respectively,was also altered in AC3-/-MOE.Therefore,not only the c AMP signaling,but also the IP3 and the Shh signaling pathways were impacted in the MOE of mice with AC3 deletion.This result may help us to understand the underlying molecular mechanisms of olfactory defections in mice with AC3 deletion.
引文
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