Investigation of the regulation of transcriptional changes in Ancylostoma caninum larvae following serum activation, with a focus on the insulin-like signalling pathway
Investigation of the regulation of transcriptional changes in Ancylostoma caninum larvae following serum activation, with a focus on the insulin-like signalling pathway
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DOI：
10.1016/j.vetpar.2008.10.026
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发表时间：
2009-02-05
影响因子：
2.6
通讯作者：
Gasser, Robin B.
中科院分区：
文献类型：
--
作者：
Datu, Bennett J. D.;Loukas, Alex;Gasser, Robin B.
在自由生活的线虫秀丽隐杆线虫中，从Dauer的退出是在胰岛素样信号通路的单个双端神经元（ASJ）的控制下。该途径的突变具有抑制进入Dauer阶段的能力。据推测，胰岛素样信号在犬钩虫第三期幼虫（L3 s）体外血清刺激反应中起着重要作用。为了测试胰岛素样信号级联可能参与对血清刺激的反应，探索了两种信号兴奋剂（8-溴环鸟苷和槟榔碱）和四种抑制剂（即4，7-菲咯啉、磷酸肌醇-3激酶（PI 3 K））、Akt抑制剂IV和雷帕霉素对血清刺激的L3中选定激活相关mRNA的进食和油水平的影响。A的L3。犬用或不用适当的抑制剂/激动剂预孵育。在血清刺激后，评估摄食活性。通过逆转录实时荧光定量PCR（RTPCR）技术，研究了与特定表达序列标签（ESTs）相连的一些激活相关mRNA的转录水平。用4，7-菲咯啉治疗蠕虫完全抑制进食，并显著降低了大多数激活相关mRNA的差异水平，而用cGMP治疗导致近85%的美国人恢复进食，并产生了与血清刺激后一致的特异性转录谱。用槟榔碱处理L3导致与85%的L3相似的进食恢复，但没有导致与激活一致的转录组学特征。在PI 3 K抑制剂LY 294002（1 mM）存在下记录到进食的完全减少，并导致对所检查分子的血清刺激应答的差异转录的显著抑制。Akt抑制剂IV导致摄食减少了70%，但对所研究的任何激活相关mRNA的水平几乎没有影响。雷帕霉素显示出对进食的微弱影响，并且所研究的几种mRNA在处理后表现出大于预期的转录。激活相关转录的复杂性无法使用当前的方法来解决。为了预测或鉴定在A.犬的(C)2008由爱思唯尔公司出版
The exit from dauer in the free-living nematode Caenorhabditis elegans is under the control of a single amphidial neuron (ASJ) of the insulin-like signalling pathway. Mutations of this pathway have the ability to Suppress entry into the dauer stage. It has been Postulated that insulin-like signalling plays a significant role in the response to serum stimulation in vitro of the third-stage larvae (L3s) of the canine hookworm Ancylostoma caninum. To test for the possible involvement of the insulin-like signalling cascade in the response to serum stimulation, the effects of two signalling stimulants (8-bromo cGMP and arecoline) and four inhibitors, namely 4,7-phenanthroline, phosphoinositide-3 kinase (PI3K), Akt inhibitor IV and rapamycin on feeding and oil levels of selected activation-associated mRNAs in serum-stimulated L3s were explored. L3s of A. caninum Were pre-incubated with or Without the appropriate inhibitor/agonist. Following serum-stimulation, the feeding activity was assessed. The transcription levels of a number of activation-associated mRNAs linked to particular expressed sequences tags (ESTs) were investigated by reverse transcription, real-time PCR (rtPCR). The treatment of worms with 4,7-phenanthroline completely suppressed feeding and significantly reduced the differential levels of most activation-associated mRNAs, whereas the treatment with cGMP resulted in the resumption of feeding in almost 85% of the Us and yielded a specific transcriptional profile consistent with that following serum stimulation. The treatment of L3s with arecoline resulted in the resumption of feeding in similar to 85% of L3s, but did not result in a transcriptomic profile consistent with activation. A complete reduction in feeding was recorded in the presence of the PI3K inhibitor LY294002 (1 mM) and resulted in a pronounced dampening of differential transcription in response to serum stimulation for the molecules examined. Akt inhibitor IV resulted in a similar to 70% reduction in feeding but had almost no effect on the level of any of the activation-associated mRNAs Studied. Rapamycin was shown to have a weak effect on feeding, and several of the mRNAs Studied exhibited greater than expected transcription following treatment. The complexities of activation-associated transcription could not be addressed using the current approach. A larger number of mRNAs needs to be investigated in order to predict or identify regulatory mechanisms proposed to function in the insulin-like signalling pathway in A. caninum. (C) 2008 Published by Elsevier B.V.