Matsuoka Hiroyuki, Tomita Hiroyuki, Hattori Ryuta, Arai Meiji, Hirai Makoto
Division of Medical Zoology, Jichi Medical University , 3311-1 Yakushiji, Shimotsuke-shi 329-0498, Japan.
Division of Medical Zoology, Jichi Medical University , 3311-1 Yakushiji, Shimotsuke-shi 329-0498, Japan ; Department of International Medical Zoology, Graduate School of Medicine, Kagawa University , Miki-cho 761-0793, Japan.
Trop Med Health. 2015 Mar;43(1):53-61. doi: 10.2149/tmh.2014-18. Epub 2014 Nov 20.
We produced a transgenic rodent malaria parasite (Plasmodium berghei) that contained the luciferase gene under a promoter region of elongation factor-1α. These transgenic (TG) parasites expressed luciferase in all stages of their life cycle, as previously reported. However, we were the first to succeed in observing sporozoites as a mass in mouse skin following their deposition by the probing of infective mosquitoes. Our transgenic parasites may have emitted stronger bioluminescence than previous TG parasites. The estimated number of injected sporozoites by mosquitoes was between 34 and 775 (median 80). Since luciferase activity diminished immediately after the death of the parasites, luciferase activity could be an indicator of the existence of live parasites. Our results indicated that sporozoites survived at the probed site for more than 42 hours. We also detected sporozoites in the liver within 15 min of the intravenous injection. Bioluminescence was not observed in the lung, kidney or spleen. We confirmed the observation that the liver was the first organ in which malaria parasites entered and increased in number.
我们构建了一种转基因啮齿动物疟原虫(伯氏疟原虫),其在延伸因子-1α的启动子区域下含有荧光素酶基因。如先前报道的那样,这些转基因(TG)寄生虫在其生命周期的所有阶段均表达荧光素酶。然而,我们是首个成功观察到感染性蚊子叮咬后,子孢子在小鼠皮肤中聚集的情况。我们的转基因寄生虫可能比先前的TG寄生虫发出更强的生物发光。蚊子注射的子孢子估计数量在34至775之间(中位数为80)。由于寄生虫死亡后荧光素酶活性立即减弱,荧光素酶活性可能是活寄生虫存在的指标。我们的结果表明,子孢子在叮咬部位存活超过42小时。我们还在静脉注射后15分钟内在肝脏中检测到子孢子。在肺、肾或脾中未观察到生物发光。我们证实了肝脏是疟原虫进入并数量增加的首个器官这一观察结果。