Gene discovery and differential expression analysis of humoral immune response elements in female Culicoides sonorensis (Diptera: Ceratopogonidae)

Particle and Fibre Toxicology

Christopher A Saski¹  Matthew B Lee²  Dana Nayduch² 

[1] Clemson University Genomics and Computational Biology Laboratory, Institute for Translational Genomics, BRC #310, 105 Collins St, Clemson, SC 29634, USA;USDA-ARS, Center for Grain and Animal Health Research, Arthropod Borne Animal Diseases Research Unit, 1515 College Avenue, Manhattan, KS 66502, USA

关键词: Defensin;    Cecropin;    Attacin;    RNAseq;    JAK/STAT;    Imd;    Toll;    Biting midge;    Innate immunity;   

Others  :  1181848
DOI  :  10.1186/1756-3305-7-388

 received in 2014-06-09, accepted in 2014-08-01,  发布年份 2014

【 摘 要 】

Background

Female Culicoides sonorensis midges (Diptera: Ceratopogonidae) are vectors of pathogens that impact livestock and wildlife in the United States. Little is known about their biology on a molecular-genetic level, including components of their immune system. Because the insect immune response is involved with important processes such as gut microbial homeostasis and vector competence, our aims were to identify components of the midge innate immune system and examine their expression profiles in response to diet across time.

Methods

In our previous work, we de novo sequenced and analyzed the transcriptional landscape of female midges under several feeding states including teneral (unfed) and early and late time points after blood and sucrose. Here, those transcriptomes were further analyzed to identify insect innate immune orthologs, particularly humoral immune response elements. Additionally, we examined immune gene expression profiles in response to diet over time, on both a transcriptome-wide, whole-midge level and more specifically via qRTPCR analysis of antimicrobial peptide (AMP) expression in the alimentary canal.

Results

We identified functional units comprising the immune deficiency (Imd), Toll and JAK/STAT pathways, including humoral factors, transmembrane receptors, signaling components, transcription factors/regulators and effectors such as AMPs. Feeding altered the expression of receptors, regulators, AMPs, prophenoloxidase and thioester-containing proteins, where blood had a greater effect than sucrose on the expression profiles of most innate immune components. qRTPCR of AMP genes showed that all five were significantly upregulated in the alimentary canal after blood feeding, possibly in response to proliferating populations of gut bacteria.

Conclusions

Identification and functional insight of humoral/innate immune components in female C. sonorensis updates our knowledge of the molecular biology of this important vector. Because diet alone influenced the expression of immune pathway components, including their effectors, subsequent study of the role of innate immunity in biological processes such as gut homeostasis and life history are being pursued. Furthermore, since the humoral response is a key contributor in gut immunity, manipulating immune gene expression will help in uncovering genetic components of vector competence, including midgut barriers to infection. The results of such studies will serve as a platform for designing novel transmission-blocking strategies.

【 授权许可】

   
2014 Nayduch et al.; licensee BioMed Central Ltd.

【 图 表 】

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