Abstract

BackgroundThe fat body is the main organ of intermediary metabolism in insects and the principal source of hemolymph proteins. As part of our ongoing efforts to understand mosquito fat body physiology and to identify novel targets for insect control, we have conducted a transcriptome analysis of the fat body of Aedes aegypti before and in response to blood feeding.ResultsWe created two fat body non-normalized EST libraries, one from mosquito fat bodies non-blood fed (NBF) and another from mosquitoes 24 hrs post-blood meal (PBM). 454 pyrosequencing of the non-normalized libraries resulted in 204,578 useable reads from the NBF sample and 323,474 useable reads from the PBM sample. Alignment of reads to the existing reference Ae. aegypti transcript libraries for analysis of differential expression between NBF and PBM samples revealed 116,912 and 115,051 matches, respectively. De novo assembly of the reads from the NBF sample resulted in 15,456 contigs, and assembly of the reads from the PBM sample resulted in 15,010 contigs. Collectively, 123 novel transcripts were identified within these contigs. Prominently expressed transcripts in the NBF fat body library were represented by transcripts encoding ribosomal proteins. Thirty-five point four percent of all reads in the PBM library were represented by transcripts that encode yolk proteins. The most highly expressed were transcripts encoding members of the cathepsin b, vitellogenin, vitellogenic carboxypeptidase, and vitelline membrane protein families.ConclusionThe two fat body transcriptomes were considerably different from each other in terms of transcript expression in terms of abundances of transcripts and genes expressed. They reflect the physiological shift of the pre-feeding fat body from a resting state to vitellogenic gene expression after feeding.

Highlights

  • The yellow fever mosquito, Aedes aegypti, is the primary vector for dengue fever, several encephalitis viruses, yellow fever, and several types of filariasis [1]

  • The fat body is the principal organ of intermediary metabolism, functioning as a storage unit for lipids, carbohydrates, and proteins in both mosquitoes and insects in general

  • For practical reasons we isolated abdominal body walls with attached abdominal fat body tissue and processed them for library construction. This is a standard preparation for studying mosquito fat body physiology [14,18,23]

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Summary

Introduction

The yellow fever mosquito, Aedes aegypti, is the primary vector for dengue fever, several encephalitis viruses, yellow fever, and several types of filariasis [1]. The fat body is the principal organ of intermediary metabolism, functioning as a storage unit for lipids, carbohydrates, and proteins in both mosquitoes and insects in general. The principal cell type of the fat body is a large polyploid cell referred to as the trophocyte, which is capable of synthesizing large amounts of protein and contains many ribosomes and oil droplets [3]. The fat body acts as the main source of hemolymph proteins in all developmental stages of holo-, hemi-, and a-metabolic insects [4]. The fat body is the main organ of intermediary metabolism in insects and the principal source of hemolymph proteins. As part of our ongoing efforts to understand mosquito fat body physiology and to identify novel targets for insect control, we have conducted a transcriptome analysis of the fat body of Aedes aegypti before and in response to blood feeding

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