An improved method for the in vitro differentiation of Plasmodium falciparum gametocytes into ookinetes.
An improved method for the in vitro differentiation of Plasmodium falciparum gametocytes into ookinetes.
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DOI：
10.1186/1475-2875-9-194
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发表时间：
2010-07-08
期刊：
影响因子：
3
通讯作者：
Jacobs-Lorena M
中科院分区：
文献类型：
--
作者：
Ghosh AK;Dinglasan RR;Ikadai H;Jacobs-Lorena M
Ookinete 是疟疾寄生虫的一种形式，它侵入蚊子中肠上皮以启动孢子生殖。摄入的配子细胞在蚊子中肠腔中分化为动动子，并随后与中肠上皮的腔表面相互作用，为入侵做准备，这是一个复杂且多步骤的过程。为了促进对这些事件的详细研究，有必要产生足够数量的纯的、完全成熟的和有功能的动动子。然而，即使是少量的恶性疟原虫动合子的体外生产也已被证明是一项艰巨的任务。因此，在过去的四十年里，我们对这种寄生虫的生物学的集体理解仍然严重不足。本文报道了对五种不同动动子培养基的研究，旨在提高恶性疟原虫配子细胞体外转化为成熟动动子的效率及其对蚊子中肠的感染性。评估了五种不同的动动体培养基支持配子细胞分化为配子并进一步分化为成熟的第五阶段动动体的能力。此外，通过将体外转化的动动子喂给媒介蚊子并测量它们穿过中肠并形成卵囊的能力来评估体外转化的动动子的感染性。五种培养基中的一种（E培养基）显然更优越，因为培养的动动物在喂给蚊子时产生最多数量的卵囊。关键成分是添加了人血清、人红细胞裂解物和蚊蛹提取物，从而产生大量动合子，当喂给蚊子时能够发育成卵囊。这种对现行方法的简单而实用的改进将有助于研究这种重要的人类疟疾寄生虫如何在蚊子中开始发育，并将有助于了解其传播生物学。
Ookinete is the form of the malaria parasite that invades the mosquito midgut epithelium to initiate sporogony. Differentiation of ingested gametocytes into ookinetes in the mosquito midgut lumen and subsequent interaction with the lumenal surface of the midgut epithelium in preparation for invasion is a complex and multi-stepped process. To facilitate the study of these events in detail it is necessary to produce sufficient numbers of pure, fully mature and functional ookinetes. However, production of even a small number of Plasmodium falciparum ookinetes in vitro has proven to be a daunting task. Consequently, over the past four decades our collective understanding of the biology of this parasite form remains sorely deficient. This article reports on investigations of five different ookinete media, in an effort to improve the in vitro transformation efficiency of P. falciparum gametocytes into mature ookinetes and their infectivity of the mosquito midgut. Five different ookinete media were evaluated for their ability to support the differentiation of gametocytes into gametes and further into mature stage V ookinetes. Moreover, infectivity of the in vitro-transformed ookinetes was evaluated by feeding them to vector mosquitoes and measuring their ability to traverse the midgut and form oocysts. One of the five media (medium E) was clearly superior in that the cultured ookinetes produced the largest number of oocysts when fed to mosquitoes. Key components were additions of human serum, human red blood cell lysate and mosquito pupal extract, resulting in the production of larger numbers of ookinetes able to develop into oocysts when fed to mosquitoes. This simple and practical improvement over the prevailing methodology will facilitate the investigation of how this important human malaria parasite initiates its development in the mosquito and will contribute to the understanding of its transmission biology.
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