Functional study of acetylcholinesterase genes in Bombyx mori ovary cells using RNA interference
Functional study of acetylcholinesterase genes in Bombyx mori ovary cells using RNA interference
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利用RNA干扰技术研究家蚕卵巢细胞乙酰胆碱酯酶基因的功能
DOI：
10.1111/j.1570-7458.2011.01207.x
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发表时间：
2012-02-01
影响因子：
1.9
通讯作者：
Li, B.
中科院分区：
文献类型：
--
作者：
Cao, Y. Q.;Liu, L. H.;Li, B.
乙酰胆碱酯酶（Acetylcholinesterase，AChE）在神经传递中起着关键作用，它通过催化神经元突触中的乙酰胆碱来维持神经冲动的传递。鳞翅目昆虫乙酰胆碱酯酶由两个ace基因ace 1和ace 2编码。在这项研究中，我们使用RNA干扰分别敲低ace 1或ace 2在家蚕（L.）（鳞翅目：鳞翅目）细胞系（BmN），探讨其功能及相互关系。使用定量实时PCR确认敲低效率。我们还监测了B后不同时间点ace 1和ace 2的转录水平。家蚕核型多角体病毒（BmNPV）感染。我们发现当ace 1的表达被敲低到37.5%时，ace 2的表达增加了66%;当ace 2的表达被敲低到33.4%时，ace 1的表达同样被敲低到26.7%。当ace 2表达被抑制时，乙酰胆碱酯酶活性下降到86.2%。然而，没有观察到显着差异时，乙酰胆碱酯酶活性ace 1表达被敲低。感染BmNPV后，ace 1表达逐渐增强，ace 2表达逐渐减弱。我们的研究表明，ace 2起着重要的补偿作用的ace 1的损失，在维持AChE活性，通过增加其表达水平。此外，ace 2还可能调控ace 1的转录。总的来说，我们的研究结果有助于阐明鳞翅目中两个ace基因之间的关系。
Acetylcholinesterase (AChE) plays a key role in neurotransmission, in which it maintains the transmission of nerve impulses by catalyzing acetylcholine in the neuronal synapse. Acetylcholinesterase in Lepidoptera is encoded by two ace genes, ace1 and ace2. In this study, we used RNA interference to individually knock down the expression of ace1 or ace2 in a Bombyx mori (L.) (Lepidoptera: Bombycidae) cell line (BmN) to explore their functions and relationships. Knockdown efficiency was confirmed using quantitative real-time PCR. We also monitored transcription levels of ace1 and ace2 at different time points after B. mori nucleopolyhedrovirus (BmNPV) infection. We found that when the expression of ace1 was knocked down to 37.5%, the expression of ace2 increased by 66%; when ace2 expression was reduced to 33.4%, the ace1 expression was similarly reduced to 26.7%. Acetylcholinesterase activity was decreased to 86.2% when ace2 expression was inhibited. However, no significant difference was observed in AChE activity when ace1 expression was knocked down. After BmNPV infection, ace1 expression was gradually increasing, whereas ace2 expression was gradually decreasing. Our study indicates that ace2 plays an important compensatory role for the loss of ace1 in maintaining AChE activity by increasing its expression level. In addition, ace2 may also regulate transcription of ace1. Overall, our findings help elucidate the relationship between the two ace genes in Lepidoptera.