Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2009 Aug;81(2):258-63.

West Nile virus infection alters midgut gene expression in Culex pipiens quinquefasciatus Say (Diptera: Culicidae)

Chelsea T Smartt  1 , Stephanie L Richards, Sheri L Anderson, Jennifer S Erickson
Affiliations

West Nile virus infection alters midgut gene expression in Culex pipiens quinquefasciatus Say (Diptera: Culicidae)

Chelsea T Smartt et al. Am J Trop Med Hyg. 2009 Aug.

Abstract

Alterations in gene expression in the midgut of female Culex pipiens quinquefasciatus exposed to blood meals containing 6.8 logs plaque-forming units/mL of West Nile virus (WNV) were studied by fluorescent differential display. Twenty-six different cDNAs exhibited reproducible differences after feeding on infected blood. Of these, 21 cDNAs showed an increase in expression, and 5 showed a decrease in expression as a result of WNV presence in the blood meal. GenBank database searches showed that one clone with increased expression, CQ G12A2, shares 94% identity with a leucine-rich repeat-containing protein from Cx. p. quinquefasciatus and 32% identity to Toll-like receptors from Aedes aegypti. We present the first cDNA clone isolated from female Cx. p. quinquefasciatus midgut tissue whose expression changes on exposure to WNV. This cDNA represents a mosquito gene that is an excellent candidate for interacting with WNV in Cx. p. quinquefasciatus and may play a role in disease transmission.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Results from the fluorescent differential display analysis of the gene expression differences between midgut tissue isolated from Cx. p. quinquefasciatus female mosquitoes 4 days after ingestion of uninfected (Sample 1) and WNV-infected (Sample 2) blood. Numbers 1–14 at the top of the figure represent 14 of the 48 primers used in the DD reaction. Each primer was used in a PCR reaction with RNA from uninfected midguts (Sample 1, first two lanes) and infected midguts (Sample 2, next two lanes). Each reaction was duplicated, as shown on the figure as Samples 1′ and 2′. The red arrows indicate those products with differential gene expression. The black circle points out the expression of the product CQ G12A resulting from fluorescent differential display.
Figure 2
Figure 2
Multiple protein sequence alignment of a portion of the Cx. p. quinquefasciatus CQ G12A2 putative translation product (94 amino acids; CQ G12A2 protein; accession no. GO254244), with partial sequences representing the Toll protein (AE Toll protein; accession no. XP_001650338.1) and leucine-rich repeat protein (CQ leucine-rich repeat protein; accession no. XP_001846467.1) from Ae. aegypti and Cx. p. quinquefasciatus, respectively. The numbering represents the amino acid number. The amino acids that are underlined represent the putative conserved region containing leucine repeats in the CQ G12A2 translation product. The boxed residues represent the hydrophobic amino acids contained in the underlined region. The stars indicate leucine residues conserved in all three aligned amino acid sequences. LRR, leucine-rich repeat region.
Figure 3
Figure 3
Semi-quantitative RT-PCR analyses of DD clone CQ G12A2 in Cx. p. quinquefasciatus mosquito midgut RNA isolated from females fed an uninfected blood meal (A). B, Integrity of the RNA. Bp, base pairs; h, hours; d, days.
Figure 4
Figure 4
Semi-quantitative RT-PCR analyses of DD clone CQ G12A2 in Cx. p. quinquefasciatus mosquito midgut RNA isolated from females fed a WNV-infected blood meal (A). B, Agarose gel of the qRT-PCR analysis of WNV titer in the same RNA used in A. C, Integrity of the RNA. Bp, base pairs; h, hours; d, days.
Figure 5
Figure 5
Culex p. quinquefasciatus midgut WNV titer at different time points after infection. When analyzed using analysis of variance, there is a significant difference in WNV titer in the mosquito midgut at different times after infection (F = 70.54, df = 12.38, P ≤ 0.001). Means with the same letter are not significantly different using the Duncan multiple range test.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Murgue B, Murri S, Zientara S, Durand B, Durand JP, Zeller H. West Nile outbreak in horses in southern France, 2000: the return after 35 years. Emerg Infect Dis. 2001;7:692–696. - PMC - PubMed
    1. Autorino GL, Battisti A, Deubel V, Ferrari G, Forletta R, Giovannini A, Lelli R, Murri S, Scicluna MT. West Nile virus epidemic in horses, Tuscany region, Italy. Emerg Infect Dis. 2002;8:1372–1378. - PMC - PubMed
    1. Peyrefitte CN, Pastorino B, Grau GE, Lou J, Tolou H, Couissinier-Paris P. Dengue virus infection of human microvascular endothelial cells from different vascular beds promotes both common and specific functional changes. J Med Virol. 2006;78:229–242. - PubMed
    1. Kramer LD, Styer LM, Ebel GD. A global perspective on the epidemiology of West Nile virus. Annu Rev Entomol. 2008;53:61–81. - PubMed
    1. Sardelis MR, Turell MJ, Dohm DJ, O'guinn ML. Vector competence of selected North American Culex and Coquillettidia mosquitoes for West Nile virus. Emerg Infect Dis. 2001;7:1018–1022. - PMC - PubMed

Publication types

LinkOut - more resources