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腺苷酸环化酶3(Adenylate cyclase 3,AC3)是膜整合蛋白,可将ATP变成cAMP,使cAMP信号通路受损,进而导致嗅觉丧失。主要嗅觉表皮(main olfactory epithelium,MOE)组织是感知和传导嗅觉信号的重要器官,主要通过环磷酸腺苷cAMP、肌醇1,4,5-三磷酸(Inositol 1,4,5-trisphosphate,IP3)、刺猬(Hedgehog,Hh)等3个信号通路进行嗅觉信号转导。MOE组织中有AC2、AC3、AC4等表达,为探讨当AC3缺失时,AC2和AC4是否可以发挥AC3作用介导嗅觉信号转导;AC3缺失是否通过引起IP3和Hh信号通路的改变而介导cAMP信号通路,本研究拟选取AC3缺失的小鼠和野生型小鼠各4只,以小鼠的MOE组织为实验材料,利用实时荧光定量PCR(q PCR)技术检测小鼠MOE组织内的AC2、AC4;检测IP3信号通路中的ip3r1、calm1、calm2;检测Hh信号通路中的patched、smoothened、gli1和gli2等基因的表达情况。结果显示,AC3缺失的小鼠中AC3几乎无表达;和野生型小鼠相比,AC3缺失的小鼠MOE组织中AC2、AC4表达量均显著下降(p<0.05),分别下调1倍和4倍;AC3缺失的小鼠IP3信号通路中的三磷酸肌醇Ⅰ型受体(type 1 inositol1,4,5-trisphosphate receptor,ip3r1)、calm1、calm2表达量均显著下降(p<0.05),分别下调0.3倍、10倍及4倍;AC3缺失的小鼠Hh信号通路中patched、smoothened、gli1和gli2的表达量均显著下降(p<0.05),分别下调0.4倍、10倍、10倍、4倍。因此,AC3缺失会导致cAMP信号通路的调节受损,进而影响AC3相关因子AC2和AC4的表达,同时抑制IP3和Hh信号通路的传导,在探究AC3基因敲出小鼠嗅觉丧失方面具有一定的指导意义。
Adenylate cyclase 3 (AC3) is a membrane-bound protein that turns ATP into cAMP, impairs the cAMP signaling pathway and leads to loss of smell. The main olfactory epithelium (MOE) tissue is an important organ that senses and conducts olfactory signals mainly through cyclic adenosine monophosphate (cAMP), inositol 1,4,5-trisphosphate (IP3) ), Hedgehog (Hedgehog, Hh) and other three signal pathways for olfactory signal transduction. AC2, AC3, AC4 and so on in MOE tissues. To investigate whether AC2 and AC4 can play the role of AC3 in olfactory signal transduction when AC3 is deleted, whether AC3 mediates cAMP through the change of IP3 and Hh signaling pathways In this study, four mice with AC3 deletion and four wild-type mice each were selected. MOE tissues of mice were used as experimental materials, and AC2 in mouse MOE tissues was detected by real-time fluorescence quantitative PCR (qPCR) AC4; detected ip3r1, calm1, calm2 in IP3 signal pathway; detected the expression of patched, smoothened, gli1 and gli2 genes in Hh signaling pathway. AC3-deficient mice showed almost no expression of AC3. Compared with wild-type mice, AC2 and AC4 expression in ACE-deficient mice decreased significantly (p <0.05) and decreased by 1 and 4, respectively (P <0.05). The expression of type I inositol 1,4,5-trisphosphate receptor (ip3r1), calm1 and calm2 in IP3 signaling pathway of AC3-deficient mice were significantly decreased Down-regulated 0.3, 10 and 4 times respectively. The expression of patched, smoothened, gli1 and gli2 in the Hh signaling pathway of AC3-deficient mice were significantly decreased (p <0.05) Times Therefore, deletion of AC3 leads to the impaired regulation of cAMP signaling pathway, which in turn affects the expression of AC3-related factors AC2 and AC4, as well as the conduction of IP3 and Hh signaling pathways, and has some guidance in exploring the loss of olfactory function in AC3 knockout mice significance.