Abstract
Anopheline mosquitoes are major vectors of malaria parasites. When the gametocytes of the malaria parasite are transferred from a vertebrate to mosquitoes, they differentiate into gametes, and are fertilized in the midguts of mosquitoes. Xanthurenic acid (XA), a waste product of the ommochrome synthesis pathway, has been shown to induce exflagellation during microgametogenesis in vitro; however, it currently remains unclear whether endogenous XA affects the infectivity of anopheline mosquitoes to malaria parasites in vivo due to the lack of appropriate experimental systems such as a XA-deficient line. In the present study, we produced a XA-deficient line in Anopheles stephensi using transcription activator-like effector nuclease (TALEN)-mediated gene targeting (knockout) of the kynurenine 3-monooxygenase (kmo) gene, which encodes an enzyme that participates in the ommochrome synthesis pathway. The knockout of kmo resulted in the absence of XA, and oocyst formation was inhibited in the midguts of these XA-deficient mosquitoes, which, in turn, reduced sporozoite numbers in their salivary glands. These results suggest that endogenous XA stimulates exflagellation, and enhances the infectivity of anopheline mosquitoes to malaria parasites in vivo. The XA-deficient line of the anopheline mosquito provides a useful system for analyzing and understanding the associated factors of malaria gametogenesis in the mosquito midgut.
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Acknowledgements
We are grateful to C. Seki (Jichi Medical University) for the excellent assistance with the dissection of mosquitoes. We thank K. Watano, J. Sato, and Y. Hata (Jichi Medical University) for rearing the mosquitoes and mice. We also thank Y. Takasu (National Agriculture and Food Research Organization) for providing the pBlue-TAL plasmids. This work was funded by grants from The Science Research Promotion Fund from the Promotion and Mutual Aid Corporation for Private School of Japan to DSY, Takeda Science Foundation to DSY, JSPS KAKENHI Grant Number 16K18824 to DSY, and JSPS KAKENHI Grant Number 17K08161 to MS.
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Yamamoto, D.S., Sumitani, M., Hatakeyama, M. et al. Malaria infectivity of xanthurenic acid-deficient anopheline mosquitoes produced by TALEN-mediated targeted mutagenesis. Transgenic Res 27, 51–60 (2018). https://doi.org/10.1007/s11248-018-0057-2
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DOI: https://doi.org/10.1007/s11248-018-0057-2