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Nix is a male-determining factor in the Asian tiger mosquito Aedes albopictus.

ABSTRACT: The initial signal that governs sex determination is highly variable among insects. A homolog of Nix, the male-determining factor in Aedes aegypti, was previously found in the Asian tiger mosquito Ae. albopictus. Here we show that the Ae. albopictus Nix (AalNix) is more complex in gene structure and splice isoforms than its Ae. aegypti homolog (AaeNix). AalNix shows a similar transcription profile compared to AaeNix. CRISPR/Cas9-mediated knockouts of AalNix in vivo and in the Ae. albopictus C6/36 cells lead to a shift of dsx and fru splicing towards the female isoforms. G₀ knockout males showed feminization and deformities including feminized antennae, absence or partial absence of gonocoxites, gonostyli, testes and accessory glands, and the formation of ovaries. Despite ~70 MY of divergence, Nix functions as a conserved male-determining factor in the two most important arboviral vectors, namely Ae. aegypti and Ae. albopictus.

SUBMITTER: Liu P

PROVIDER: S-EPMC10211468 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

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Nix is a male-determining factor in the Asian tiger mosquito Aedes albopictus.

Liu Peiwen P Jin Binbin B Li Xiaocong X Zhao Yijie Y Gu Jinbao J Biedler James K JK Tu Zhijian Jake ZJ Chen Xiao-Guang XG

Insect biochemistry and molecular biology 20191231

The initial signal that governs sex determination is highly variable among insects. A homolog of Nix, the male-determining factor in Aedes aegypti, was previously found in the Asian tiger mosquito Ae. albopictus. Here we show that the Ae. albopictus Nix (AalNix) is more complex in gene structure and splice isoforms than its Ae. aegypti homolog (AaeNix). AalNix shows a similar transcription profile compared to AaeNix. CRISPR/Cas9-mediated knockouts of AalNix in vivo and in the Ae. albopictus C6/3 ...[more]

PMID: 31901476

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Project description:BackgroundTemporal and stage specific expression of microRNAs (miRNAs) in embryos, larvae, pupae and adults of Aedes albopictus showed differential expression levels across the four developmental stages, indicating their potential regulatory roles in mosquito development. The functional characterization of these miRNAs was not known. Accordingly our study evaluated the functional characterization of three miRNAs, which are temporally up-regulated in the various developmental stages of Ae. albopictus mosquitoes.MethodsmiRNA mimics, inhibitors and negative controls were designed and their knock-in and knock-down efficiency were analyzed by qRT-PCR after transfecting the mosquito cell lines C6/36, and also by injecting in their specific developmental stages. The functional role of each individual miRNA was analyzed with various parameters of development such as, hatching rate and hatching time in embryos, eclosion rate in larvae, longevity and fecundity in the adult mosquitoes.ResultsThe knock-in with the specifically designed miRNA mimics showed increased levels of expression of miRNA compared with their normal controls. We confirmed these findings using qRT-PCR, both by in vitro expression in C6/36 mosquito cell lines after transfection as well as in in vivo expression in developmental stages of mosquitoes by microinjection. The knock-down of expression with the corresponding inhibitors showed a considerable decrease in the expression levels of these miRNAs and obvious functional effects in Ae. albopictus development, detected by a decrease in the hatching rate of embryos and eclosion rate in larvae and a marked reduction in longevity and fecundity in adults.ConclusionThis study carried out by knock-in and knock-down of specifically and temporally expressed miRNAs in Ae. albopictus by microinjection is a novel study to delineate the importance of the miRNA expression in regulating mosquito development. The knock-down and loss of function of endogenously expressed miRNAs by the miRNA inhibitors in specific developmental stages had considerable effects on development, but enhancement of their gain of function was not observed on knock-in of these specific miRNAs. Hence, our study indicates that an optimal level of endogenous expression of miRNA is indispensable for the normal development and maintenance of the vectorial population density and pathogen transmissibility of this mosquito vector.

Dataset Information

Nix is a male-determining factor in the Asian tiger mosquito Aedes albopictus.

Publications

Nix is a male-determining factor in the Asian tiger mosquito Aedes albopictus.

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