Abstract
Blood feeding is an integral process required for physiological functions and propagation of the malaria vector Anopheles. During blood feeding, presence of the malaria parasite, Plasmodium in the blood induces several host effector molecules including microRNAs which play important roles in the development and maturation of the parasite within the mosquito. The present study was undertaken to elucidate the dynamic expression of miRNAs during gonotrophic cycle and parasite development in Anopheles stephensi. Using next generation sequencing technology, we identified 126 miRNAs of which 17 were novel miRNAs. The miRNAs were further validated by northern hybridization and cloning. Blood feeding and parasitized blood feeding in the mosquitoes revealed regulation of 13 and 16 miRNAs respectively. Expression profiling of these miRNAs revealed that significant miRNAs were down-regulated upon parasitized blood feeding with a repertoire of miRNAs showing stage specific up-regulation. Expression profiles of significantly modulated miRNAs were further validated by real time PCR. Target prediction of regulated miRNAs revealed overlapping targeting by different miRNAs. These targets included several metabolic pathways including metabolic, redox homeostasis and protein processing machinery components. Our analysis revealed tight regulation of specific miRNAs post blood feeding and parasite infection in An. stephensi. Such regulated expression suggests possible role of these miRNAs during gonotrophic cycle in mosquito. Another set of miRNAs were also significantly regulated at 42 h and 5 days post infection indicating parasite stage-specific role of host miRNAs. This study will result in better understanding of the role of miRNAs during gonotrophic cycle and parasite development in mosquito and can probably facilitate in devising novel malaria control strategies at vector level.
Highlights
Vector transmitted diseases are widespread in tropics and subtropics
Anopheles stephensi is considered as an important vector with its distribution throughout the Middle East and South Asia region
Small RNA sequencing using Illumina Truseq chemistry was performed on An. stephensi mosquito samples under five different conditions
Summary
Vector transmitted diseases are widespread in tropics and subtropics. Of several vector transmitted diseases, malaria accounts for more than 200 million cases with India contributing to almost 70% of malarial cases in South East Asia region [1]. Malaria was brought on the verge of eradication by the success of malaria eradication program by World Health Organisation in 1950s. 1990s saw resurgence in malarial incidence due to insecticide resistance in vectors, drug resistance in parasites, changes in vector behaviour and lack of infrastructure to fight the disease. India has an intricate vector dynamics with six primary vectors of the 58 Anopheles species reported in the country [2]. Anopheles stephensi is considered as an important vector with its distribution throughout the Middle East and South Asia region
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