Hatta Takeshi, Umemiya Rika, Liao Min, Gong Haiyan, Harnnoi Thasaneeya, Tanaka Miho, Miyoshi Takeharu, Boldbaatar Damdinsuren, Battsetseg Badgar, Zhou Jinlin, Xuan Xuenan, Tsuji Naotoshi, Taylor Demar, Fujisaki Kozo
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido, 080-8555, Japan.
Parasitol Res. 2007 Mar;100(4):847-54. doi: 10.1007/s00436-006-0336-3. Epub 2006 Nov 29.
Ticks are effective vectors of pathogens because of their blood feeding and high fecundity. This high fecundity is related to the size of the blood meal. Therefore, knowledge of how blood proteins are degraded and converted to proteins, including yolk protein, is important for the development of ways to inhibit the utilization of blood proteins by ticks. RNA interference (RNAi) is becoming a powerful post-transcriptional gene silencing technique that provides insight into gene function. We constructed a double-stranded RNA (dsRNA) based on a previously cloned Haemaphysalis longicornis leucine aminopeptidase (HlLAP) gene to reevaluate the biological role in tick blood digestion. Gene specific transcriptional, translational, and functional disruptions were achieved by the introduction of dsRNA into the ticks. Significantly delayed onset of egg-laying and reduced egg oviposition resulted from the RNAi for the HlLAP gene. These results suggest that HlLAP actually works as a blood digestive enzyme and affects tick fecundity via unknown mechanisms. The reduction of egg oviposition may be caused by a decrease in nutrients, especially free amino acids generated by HlLAP, from the blood meal. This is the first report of an impact on tick reproduction caused by gene silencing of a blood digestion-related molecule.
蜱虫因其吸食血液和高繁殖力而成为病原体的有效传播媒介。这种高繁殖力与血餐的大小有关。因此,了解血液蛋白质如何被降解并转化为包括卵黄蛋白在内的蛋白质,对于开发抑制蜱虫利用血液蛋白质的方法至关重要。RNA干扰(RNAi)正成为一种强大的转录后基因沉默技术,有助于深入了解基因功能。我们基于先前克隆的长角血蜱亮氨酸氨肽酶(HlLAP)基因构建了双链RNA(dsRNA),以重新评估其在蜱虫血液消化中的生物学作用。通过将dsRNA引入蜱虫,实现了基因特异性的转录、翻译和功能破坏。HlLAP基因的RNA干扰导致产卵开始显著延迟和产卵量减少。这些结果表明,HlLAP实际上作为一种血液消化酶发挥作用,并通过未知机制影响蜱虫的繁殖力。产卵量的减少可能是由于血餐中营养物质的减少,特别是HlLAP产生的游离氨基酸的减少。这是关于血液消化相关分子的基因沉默对蜱虫繁殖产生影响的首次报道。