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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]
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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

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