Comparative expression profile of microRNAs in Anopheles anthropophagus midgut after blood-feeding and Plasmodium infection

Wenquan Liu,Liya Cai,Qimei Wei,Lingzi Chen,Liyang Huang,Zhenhua Hao,Shaohui Liang

doi:10.1186/s13071-017-2027-6

Abstract

BackgroundAnopheles anthropophagus is one of the major vectors of malaria in Asia. MicroRNAs (miRNAs) play important roles in cell development and differentiation as well as in the cellular response to stress and infection. In a former study, we have investigated the global miRNA profiles in relation to sex in An. anthropophagus. However, the miRNAs contributing to the blood-feeding and infection with Plasmodium are still unknown.MethodsHigh-throughput sequencing was performed to identify miRNA profiles of An. anthropophagus midguts after blood-feeding and Plasmodium infection. The expression patterns of miRNA in different midgut libraries were compared based on transcripts per million reads (TPM), and further confirmed by Northern blots. Target prediction and pathway analysis were carried out to investigate the role of regulated miRNAs in blood-feeding and Plasmodium infection.ResultsWe identified 67 known and 21 novel miRNAs in all three libraries (sugar-feeding, blood-feeding and Plasmodium infection) in An. anthropophagus midguts. Comparing with the sugar-feeding, the experssion of nine (6 known and 3 novel) and ten (9 known and 1 novel) miRNAs were significantly upregulated and downregulated respectively after blood-feeding (P < 0.05, fold change ≥ 2 and TPM ≥ 10). Plasmodium infection induced the expression of thirteen (9 known and 4 novel) and eleven (9 known and 2 novel) miRNAs significantly upregulated and downregulated, respectively, compared with blood-feeding. The representative upregulated miR-92a in blood-feeding and downregulated miR-275 in Plasmodium infection were further confirmed by Northern Blot. Putative targets of these regulated miRNAs were further investigated and classified into their pathways.ConclusionsThis study suggests that miRNAs are involved in the blood-feeding and Plasmodium infection in An. anthropophagus midgut. Further studies of the function of these differential expressed miRNAs will facilitate in better understanding of mosquito biology and anti-parasite immunity.

Highlights

Anopheles anthropophagus is one of the major vectors of malaria in Asia
After aligned to the known miRNA and pre-miRNA in miRBase, the miRNA reads from SF, BF and Plasmodium infection (PI) midguts which can match to the miRBase are 4.03 million, 1.88 million and 0.21million separately (Table 1)
Identification of miRNAs in midguts of An. anthropophagus A total of 67 known and 21 novel miRNAs were identified in the SF, BF and PI midgut libraries (Tables 2 and 3)

Summary

Introduction

Anopheles anthropophagus is one of the major vectors of malaria in Asia. The miRNAs contributing to the blood-feeding and infection with Plasmodium are still unknown. Malaria, caused by Plasmodium parasites, is a major threat to human health worldwide. Plasmodium spp. are transmitted to humans by the blood-feeding of Anopheles mosquitoes. Mosquitoes have developed various mechanisms to confront Plasmodium infection [3, 4]. The midgut represents the first barrier for the survival and development of Plasmodium [4,5,6]. A key concept that has emerged from recent studies is the molecular mechanisms of mosquito midgut that negatively and positively modulate the invasion of the parasite [5, 6]

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Discussion

Conclusion