Hostname: page-component-586b7cd67f-gb8f7 Total loading time: 0 Render date: 2024-11-22T23:33:08.155Z Has data issue: false hasContentIssue false
  • English
  • Français

Confocal Analysis of the Distribution and Persistence of Sindbis Virus (TaV-GFP) Infection in Midguts of Aedes aegypti Mosquitoes

Published online by Cambridge University Press:  19 March 2020

Jason J. Saredy ,
Florence Y. Chim ,
Zoë L. Lyski ,
Yani P. Ahearn  and
Doria F. Bowers
Jason J. Saredy
Affiliation:
Department of Biology, Temple University, Philadelphia, PA19122, USA
Florence Y. Chim
Affiliation:
Saft America Inc., 13575 Waterworks St., Jacksonville, FL32221, USA
Zoë L. Lyski
Affiliation:
Department of Molecular Microbiology and Immunology, Oregon Health and Science University, Portland, OR97239, USA
Yani P. Ahearn
Affiliation:
Department of Biology, University of North Florida, 1 UNF Drive, Jacksonville, FL32224, USA
Doria F. Bowers*
Affiliation:
Department of Biology, University of North Florida, 1 UNF Drive, Jacksonville, FL32224, USA
*
*Author for correspondence: D. F. Bowers, E-mail: [email protected]
Get access

Abstract

Biological transmission of arthropod-borne viruses (arboviruses) to vertebrate hosts by hematophagous insects poses a global threat because such arboviruses can result in a range of serious public health infectious diseases. Sindbis virus (SINV), the prototype Alphavirus, was used to track infections in the posterior midgut (PMG) of Aedes aegypti adult mosquitoes. Females were fed viremic blood containing a virus reporter, SINV [Thosea asigna virus-green fluorescent protein (TaV-GFP)], that leaves a fluorescent signal in infected cells. We assessed whole-mount PMGs to identify primary foci, secondary target tissues, distribution, and virus persistence. Following a viremic blood meal, PMGs were dissected and analyzed at various days of post blood-feeding. We report that virus foci indicated by GFP in midgut epithelial cells resulted in a 9.8% PMG infection and a 10.8% dissemination from these infected guts. The number of virus foci ranged from 1 to 3 per individual PMG and was more prevalent in the PMG-middle > PMG-frontal > PMG-caudal regions. SINV TaV-GFP was first observed in the PMG (primary target tissue) at 3 days post blood-feeding, was sequestered in circumscribed foci, replicated in PMG peristaltic muscles (secondary target tissue) following dissemination, and GFP was observed to persist in PMGs for 30 days postinfection.

Keywords

Alphaviruslaser confocalmosquito midgutSINV TaV-GFPvirus persistence
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 26 , Issue 2 , April 2020 , pp. 267 - 274
Copyright
Copyright © Microscopy Society of America 2020

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Bowers, DF, Abell, BA & Brown, DT (1995). Replication and tissue tropism of the alphavirus Sindbis in the mosquito Aedes albopictus. Virology 212, 1–12.CrossRefGoogle ScholarPubMed
Brown, MR, Crim, JW & Lea, AO (1986). FMRFamide- and pancreatic polypeptide-like immunoreactivity of endocrine cells in the midgut of a mosquito. Tissue & Cell 18, 419–428.CrossRefGoogle ScholarPubMed
Brown, MR, Raikhel, AS & Lea, AO (1985). Ultrastructure of midgut endocrine cells in the adult mosquito, Aedes aegypti. Tissue & Cell 17, 709–721.CrossRefGoogle ScholarPubMed
Campbell, CL, Keene, KM, Brackney, DE, Olson, KE, Blair, CD, Wilusz, J & Foy, BD (2008). Aedes aegypti uses RNA interference in defense against Sindbis virus infection. MBC Microbiol 8(47), 1–12.Google ScholarPubMed
Chalfie, M, Tu, Y, Euskirchen, G, Ward, WW & Prashert, DD (1994). Green fluorescent protein as a marker for gene expression. Science 263, 802–805.CrossRefGoogle ScholarPubMed
Chamberlain, RW & Sudia, WD (1961). Mechanism of transmission of viruses by mosquitoes. Ann Rev Entomol 6, 371–390.CrossRefGoogle ScholarPubMed
Chamberlin, RW (1980). Comparative and historical aspects of the togaviridae and flaviviridae. In Togaviridae and Flaviridae, Schleisinger, S & Schleisinger, JJ (Eds.), pp. 1–16. New York, NY: Plenum Press.Google Scholar
Ciano, KA, Saredy, JJ & Bowers, DF (2014). Heparan sulfate proteoglycan: An arbovirus attachment factor integral to mosquito salivary gland ducts. Viruses 6, 5182–5197.CrossRefGoogle ScholarPubMed
Clements, AN (1996). The Biology of Mosquitoes: Chapter 13 Structure of the Adult Alimentary Canal. London: Chapman and Hall, pp. 263–271.Google Scholar
Foy, BD, Myles, KM, Pierro, DL, Sanchez-Vargas, I, Uhilřová, M, Jindra, M, Beaty, BJ & Olson, KE (2004). Development of a new Sindbis virus transducing system and its characterization in three Culicine mosquitoes and two Lepidopteran species. Insect Mol Biol 13(1), 89–100.CrossRefGoogle ScholarPubMed
Franz, AWE, Kantor, AM, Passarelli, AL & Clem, RJ (2015). Tissue barriers to arbovirus infection in mosquitoes. Viruses 7, 3741–3767.CrossRefGoogle ScholarPubMed
Hardy, JL, Reeves, WC & Sjoren, RD (1976). Variations in the susceptibility of field and laboratory populations of Culex tarsalis to experimental infection with western equine encephalomyelitis virus. Am J Epidemol 103, 498–505.CrossRefGoogle ScholarPubMed
Khoo, CCH, Piper, J, Sanchez-Vargas, I, Olsen, KE & Franz, AWE (2010). The RNA interference pathway affects midgut infection- and escape barriers for Sindbis virus in Aedes aegypti. BMC Microbiol 10, 130.CrossRefGoogle ScholarPubMed
Klimstra, WB, Ryman, KD & Johnston, RE (1998). Adaption of Sindbis virus to BHK cells selects for use of heparan sulfate as an attachment receptor. J Virol 72(9), 7357–7366.CrossRefGoogle Scholar
Kramer, LD, Hardy, JL, Presser, SB & Houk, EJ (1981). Dissemination barriers for western equine encephalomyelitis virus in Culex tarsalis infected after ingestion of low virus doses. Am J Trop Med Hyg 30, 190–197.CrossRefGoogle Scholar
Labuda, M & Kozuch, O (1989). Amplification of arbovirus transmission by mosquito intradermal probing and interrupted feeding. Acta Virol 33(1), 63–67.Google ScholarPubMed
Li, X, Zhao, X, Fang, Y, Jiang, X, Duong, T, Fan, C, Huang, C-C & Kain, R (1998). Generation of destabilized green fluorescent protein as a transcription reporter. J Biol Chem 273(52), 34970–34975.CrossRefGoogle ScholarPubMed
Lyski, ZL (2013). Arbovirus persistence and selection of persistent variants following chronic infection in Aedine mosquitoes: A comparative study between Ae. aegypti and Ae. albopictus 30 days post infection with Sindbis virus. UNF Thesis and Dissertations. p. 430. Available at http://digitalcommons.unf.edu/etd/430/Google Scholar
Lyski, ZL, Saredy, JJ, Ciano, KA, Stem, J & Bowers, DF (2011). Blood feeding position increases success of recalcitrant mosquitoes. Vector Borne Zoonotic Dis 11(8), 1165–1171.CrossRefGoogle ScholarPubMed
McKnight, KL, Simpson, DA, Lin, S, Knott, TA, Polo, JM, Pence, DF, Johannsen, DB & Heidner, HW (1996). Deduced consensus sequence of Sindbis virus strain AR339: Mutations contained in laboratory strains which affect cell culture and in vivo phenotypes. J Virol 70(3), 1981–1989.CrossRefGoogle ScholarPubMed
Moriarty, GC, Moriarty, CM & Sternberger, LA (1973). Ultrastructural immunocytochemistry with unlabeled antibodies and the peroxidase-antiperoxidase complex. J Histochem Cytochem 21(9), 825–833.CrossRefGoogle ScholarPubMed
Okuda, K, de Almeida, F, Mortara, RA, Kriegar, H, Marinotti, O & Bijovsky, AT (2007). Cell death and regeneration in the midgut of the mosquito, Culex quinquefasciatus. J Insect Physiol 53, 1307–1315.CrossRefGoogle ScholarPubMed
Perrone, JB & Spielman, A (1985). Time and site of assembly of the peritrophic membrane of the mosquito Aedes aegypti. Cell Tissue Res 252, 473–478.Google Scholar
Pierro, DJ, Powers, EL & Olson, KE (2007). Genetic determinants of Sindbis virus strain TR339 affecting midgut infection in the mosquito Aedes aegypti. J Gen Virol 88, 1545–1554.CrossRefGoogle ScholarPubMed
Romoser, WS, Wasieloski, LP Jr, Pushko, P, Kondig, JP, Lerdthusnee, K, Neira, M & Ludwig, GV (2004). Evidence for arbovirus dissemination conduits from the mosquito (Diptera: Culicidae) midgut. J Med Entomol 41(3), 467–475.CrossRefGoogle ScholarPubMed
Sanchez-Vargas, I, Scott, JC, Poole-Smith, BK, Franz, AWE, Barbos-Solomieupe, V, Wilusz, J, Olson, KE & Blair, CD (2009). Dengue virus type 2 infections in Aedes aegypti are modulated by the mosquitoe's RNA interference pathway. PLoS Pathog 5(e1000299-10), 1371.CrossRefGoogle ScholarPubMed
Saredy, JJ & Bowers, DF (2014). Expression of TaV tagged Sindbis virus (TR339) in Aedes albopictus cell lines and adult mosquitoes. Proceed Microsc Microanal 20(S3), 1364–1365.CrossRefGoogle Scholar
Scott, TW, Chow, E, Strickman, D, Kittayapong, P, Wirth, RA, Lorenz, LH & Edman, JD (1993). Blood-feeding patterns of Aedes aegypti (Diptera: Culicidae) collected in a rural Thai village. J Med Entomol 30(5), 922–927.CrossRefGoogle Scholar
Seabaugh, RC, Olson, KE, Higgs, S, Carlson, JO & Beaty, BJ (1998). Development of a chimeric Sindbis Virus with enhanced per os infection of Aedes aegypti. Virology 243(1), 99–112.CrossRefGoogle ScholarPubMed
Sick, F, Beer, M, Kampen, H & Wernike, K (2019). Culicoides biting midges—underestimated vectors for arboviruses of public health and veterinary importance. Viruses 11, 376–395.CrossRefGoogle ScholarPubMed
Strauss, EG, Rice, CM & Strauss, JH (1984). Complete nucleotide sequence of the genomic RNA of Sindbis virus. Virology 133(1), 92–110.CrossRefGoogle ScholarPubMed
Sun, C, Gardner, CL, Watson, AM, Ryman, KD & Klimstra, WB (2014). Stable, high-level expression of reporter proteins from improved Alphavirus expression vectors to track replication and dissemination during encephalitic and arthritogenic disease. J Virol 88(23), 2035–2046.CrossRefGoogle ScholarPubMed
Taylor, RM, Hurlbut, TH, Work, JR, Kingston, JR & Frothingham, TE (1955). Sindbis virus: Newly recognized arthropod transmitted virus. Am J Trop Med Hyg 4, 844–862.CrossRefGoogle Scholar
Turrell, MJ, Gargan, TP II & Bailey, CL (1984). Replication and dissemination of Rift Valley Fever virus in Culex pipiens. Am J Trop Med Hyg 33, 176–181.CrossRefGoogle Scholar
Vo, M, Linser, PJ & Bowers, DF (2010). Organ-associated muscles in Aedes albopictus (Diptera:Culicidae) respond differentially to Sindbis virus. J Med Entomol 47(2), 215–225.CrossRefGoogle ScholarPubMed
Weaver, SC & Scott, TW (1990). Peritrophic membrane formation and cellular turnover in the midgut of Culiseta melanura (Diptera: Culicidae). J Med Entomol 27(5), 864–873.CrossRefGoogle Scholar
Zang, W, Mukhopadhyay, S, Pletnev, SV, Baker, TS, Kuhn, RJ & Rossmann, MG (2002). Placement of the structural proteins in Sindbis virus. J Virol 76(22), 11645–11658.CrossRefGoogle Scholar
Zieler, H, Garon, CF, Fischer, ER & Shahabuddin, M (2000). A tubular network associated with the brush-border surface of the Aedes aegypti midgut: Implications for pathogen transmission by mosquitoes. J Exp Biol 203, 1599–1611.Google ScholarPubMed