Culturing and Egg Collection of <i>Aedes aegypti</i>

Clemons, Anthony; Mori, Akio; Haugen, Morgan; Severson, David W.; Duman-Scheel, Molly

doi:10.1101/pdb.prot5507

Cited by 67 publications

(57 citation statements)

References 6 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Mosquitoes were reared and maintained in a 26°C environmental chamber kept at 85% relative humidity, with a 16 hour light:8 hour dark (L:D) cycle and a 30 minute crepuscular period following our standard lab protocol (Clemons et al, 2010). First instar larvae were subjected to either optimal or stressed rearing conditions as described by Ponlawat and Harrington (2007).…”

Section: Methodsmentioning

confidence: 99%

Larval stress alters dengue virus susceptibility in Aedes aegypti (L.) adult females

et al. 2017

Self Cite

View full text Add to dashboard Cite

In addition to genetic history, environmental conditions during larval stages are critical to the development, success and phenotypic fate of the Aedes aegypti mosquito. In particular, previous studies have shown a strong genotype-by-environment component to adult mosquito body size in response to optimal vs stressed larval conditions. Here, we expand upon those results by investigating the effects of larval-stage crowding and nutritional limitation on the susceptibility of a recent field isolate of Aedes aegypti to dengue virus serotype-2. Interestingly, female mosquitoes from larvae subjected to a stressed regime exhibited significantly reduced susceptibility to disseminated dengue infection 14 days post infection compared to those subjected to optimal regimes. Short term survivorship post-infected blood feeding was not significantly different. As with body size, dengue virus susceptibility of a mosquito population is determined by a combination of genetic and environmental factors and is likely maintained by balancing selection. Here, we provide evidence that under different environmental conditions, the innate immune response of field-reared mosquitoes exhibits a large range of phenotypic variability with regard to dengue virus susceptibility. Further, as with body size, our results suggest that mosquitoes reared under optimal laboratory conditions, as employed in all mosquito-pathogen studies to date, may not always be realistic proxies for natural populations.

show abstract

Section: Methodsmentioning

confidence: 99%

Larval stress alters dengue virus susceptibility in Aedes aegypti (L.) adult females

et al. 2017

Self Cite

View full text Add to dashboard Cite

show abstract

“…Maintain A. gambiae G3 and A. aegypti Liverpool IB12 strains (used in the representative studies below) or other strains of interest according to standard lab practice or as described previously 23,24 .…”

Section: Protocolmentioning

confidence: 99%

Chitosan/Interfering RNA Nanoparticle Mediated Gene Silencing in Disease Vector Mosquito Larvae

Zhang

Mysore

Flannery

et al. 2015

JoVE

Self Cite

View full text Add to dashboard Cite

SHORT ABSTRACT Here we describe a procedure for inhibiting gene function in disease vector mosquitoes through the use of chitosan/interfering RNA nanoparticles that are ingested by larvae. LONG ABSTRACT Vector mosquitoes inflict more human suffering than any other organism—and kill more than one million people each year. The mosquito genome projects facilitated research in new facets of mosquito biology, including functional genetic studies in the primary African malaria vector Anopheles gambiae and the dengue and yellow fever vector Aedes aegypti. RNA interference- (RNAi-) mediated gene silencing has been used to target genes of interest in both of these disease vector mosquito species. Here, we describe a procedure for preparation of chitosan/interfering RNA nanoparticles that are combined with food and ingested by larvae. This technically straightforward, high-throughput, and relatively inexpensive methodology, which is compatible with long double stranded RNA (dsRNA) or small interfering RNA (siRNA) molecules, has been used for the successful knockdown of a number of different genes in A. gambiae and A. aegypti larvae. Following larval feedings, knockdown, which is verified through qRT-PCR or in situ hybridization, can persist at least through the late pupal stage. This methodology may be applicable to a wide variety of mosquito and other insect species, including agricultural pests, as well as other non-model organisms. In addition to its utility in the research laboratory, in the future, chitosan, an inexpensive, non-toxic and biodegradable polymer, could potentially be utilized in the field.

show abstract

“…A. aegypti mosquitoes used in this study originated from the Second Military Medical University. Mosquitoes were maintained on a sugar solution and kept at 28°C, at 70% to 80% relative humidity, with a 12-h-light-12-h-dark photoperiod (14). For virus infection and gene knockdown, the mosquitoes were inoculated in the thorax by using an Eppendorf CellTram oil microinjector (Eppendorf, Hamburg, Germany) with 250 nl of diluted virus or dsRNA.…”

Section: Methodsmentioning

confidence: 99%

“…As with C. tritaeniorhynchus, Aedes aegypti is a potential threat for the transmission of JEV in Europe and the Americas due to globalization and climate change (1,10,11). A. aegypti is easy to cultivate and infect, and its genome has been characterized; thus, these mosquitoes are ideal for studying flavivirus pathogenesis (12)(13)(14)(15).…”

mentioning

confidence: 99%

mosGCTL-7, a C-Type Lectin Protein, Mediates Japanese Encephalitis Virus Infection in Mosquitoes

Liu

Qian

Jung

et al. 2017

J Virol

View full text Add to dashboard Cite

Japanese encephalitis virus (JEV) is an arthropod-borne flavivirus prevalent in Asia and the Western Pacific and is the leading cause of viral encephalitis. JEV is maintained in a transmission cycle between mosquitoes and vertebrate hosts, but the molecular mechanisms by which the mosquito vector participates in transmission are unclear. We investigated the expression of all C-type lectins during JEV infection in Aedes aegypti. The C-type lectin mosquito galactose-specific C-type lectin 7 (mosGCTL-7) (VectorBase accession no. AAEL002524) was significantly upregulated by JEV infection and facilitated infection in vivo and in vitro. mosGCTL-7 bound to the N-glycan at N154 on the JEV envelope protein. This recognition of viral N-glycan by mosGCTL-7 is required for JEV infection, and we found that this interaction was Ca 2ϩ dependent. After mosGCTL-7 bound to the glycan, mosPTP-1 bound to mosGCTL-7, promoting JEV entry. The viral burden in vivo and in vitro was significantly decreased by mosPTP-1 double-stranded RNA (dsRNA) treatment, and infection was abolished by anti-mosGCTL-7 antibodies. Our results indicate that the mosGCTL-7/mosPTP-1 pathway plays a key role in JEV infection in mosquitoes. An improved understanding of the mechanisms underlying flavivirus infection in mosquitoes will provide further opportunities for developing new strategies to control viral dissemination in nature.IMPORTANCE Japanese encephalitis virus is a mosquito-borne flavivirus and is the primary cause of viral encephalitis in the Asia-Pacific region. Twenty-four countries in the WHO Southeast Asia and Western Pacific regions have endemic JEV transmission, which exposes Ͼ3 billion people to the risks of infection, although JEV primarily affects children. C-type lectins are host factors that play a role in flavivirus infection in humans, swine, and other mammals. In this study, we investigated C-type lectin functions in JEV-infected Aedes aegypti and Culex pipiens pallens mosquitoes and cultured cells. JEV infection changed the expression of almost all C-type lectins in vivo and in vitro, and mosGCTL-7 bound to the JEV envelope protein via an N-glycan at N154. Cell surface mosPTP-1 interacted with the mosGCTL-7-JEV complex to facilitate virus infection in vivo and in vitro. Our findings provide further opportunities for developing new strategies to control arbovirus dissemination in nature.

show abstract

Culturing and Egg Collection of Aedes aegypti

Cited by 67 publications

References 6 publications

Larval stress alters dengue virus susceptibility in Aedes aegypti (L.) adult females

Larval stress alters dengue virus susceptibility in Aedes aegypti (L.) adult females

Chitosan/Interfering RNA Nanoparticle Mediated Gene Silencing in Disease Vector Mosquito Larvae

mosGCTL-7, a C-Type Lectin Protein, Mediates Japanese Encephalitis Virus Infection in Mosquitoes

Contact Info

Product

Resources

About