Midgut and salivary gland transcriptomes of the arbovirus vector Culicoides sonorensis (Diptera: Ceratopogonidae)

• Published in: Insect molecular biology Vol. 14; no. 2; pp. 121 - 136
• Main Authors: Campbell, C.L, Vandyke, K.A, Letchworth, G.J, Drolet, B.S, Hanekamp, T, Wilson, W.C
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Science Ltd 01-04-2005
• Subjects: Allergens - genetics; Amino Acid Sequence; Animals; Arbovirus; Arboviruses; Base Sequence; Ceratopogonidae; Ceratopogonidae - genetics; Ceratopogonidae - metabolism; Ceratopogonidae - virology; complementary DNA; Culicoides; Culicoides sonorensis; DNA Primers; DNA, Complementary - genetics; EST; expressed sequence tag; Expressed Sequence Tags; Female; Gastrointestinal Tract - metabolism; Gene Expression; genomics; gut; haematophagous; hematophagous insects; hematophagy; In Situ Hybridization; Insect Proteins - genetics; insect vectors; Insect Vectors - genetics; Insect Vectors - metabolism; Insect Vectors - virology; Male; messenger RNA; midgut; Molecular Sequence Data; Nerve Tissue Proteins - genetics; Polymerase Chain Reaction - methods; RNA, Messenger - genetics; RNA, Messenger - metabolism; salivary glands; Salivary Glands - metabolism; Sequence Alignment; Sequence Analysis, DNA; Sex Factors; transcription (genetics); vector biology
• ISSN: 0962-1075; 1365-2583
• DOI: 10.1111/j.1365-2583.2004.00537.x

Numerous Culicoides spp. are important vectors of livestock or human disease pathogens. Transcriptome information from midguts and salivary glands of adult female Culicoides sonorensis provides new insight into vector biology. Of 1719 expressed sequence tags (ESTs) from adult serum-fed female midguts harvested within 5 h of feeding, twenty-eight clusters of serine proteases were derived. Four clusters encode putative iron binding proteins (FER1, FERL, PXDL1, PXDL2), and two clusters encode metalloendopeptidases (MDP6C, MDP6D) that probably function in bloodmeal catabolism. In addition, a diverse variety of housekeeping cDNAs were identified. Selected midgut protease transcripts were analysed by quantitative real-time PCR (q-PCR): TRY1_115 and MDP6C mRNAs were induced in adult female midguts upon feeding, whereas TRY1_156 and CHYM1 were abundant in midguts both before and immediately after feeding. Of 708 salivary gland ESTs analysed, clusters representing two new classes of protein families were identified: a new class of D7 proteins and a new class of Kunitz-type protease inhibitors. Additional cDNAs representing putative immunomodulatory proteins were also identified: 5' nucleotidases, antigen 5-related proteins, a hyaluronidase, a platelet-activating factor acetylhydrolase, mucins and several immune response cDNAs. Analysis by q-PCR showed that all D7 and Kunitz domain transcripts tested were highly enriched in female heads compared with other tissues and were generally absent from males. The mRNAs of two additional protease inhibitors, TFPI1 and TFPI2, were detected in salivary glands of paraffin-embedded females by in situ hybridization.