Methods to study Drosophila immunity

Neyen, Claudine; Bretscher, Andrew J.; Binggeli, Olivier; Lemaître, Bruno

doi:10.1016/j.ymeth.2014.02.023

Cited by 127 publications

(133 citation statements)

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“…While a recent review of infection methods suggested that abdomen inoculation might be riskier due to imagined potential puncturing of the intestines, our data demonstrate that thorax inoculation is more detrimental (35). Drosophila immunity studies are somewhat divided between the two inoculation sites.…”

Section: Discussionmentioning

confidence: 57%

Thorax Injury Lowers Resistance to Infection in Drosophila melanogaster

et al. 2014

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The route of infection can profoundly affect both the progression and outcome of disease. We investigated differences in Drosophila melanogaster defense against infection after bacterial inoculation into two sites-the abdomen and the thorax. Thorax inoculation results in increased bacterial proliferation and causes high mortality within the first few days of infection. In contrast, abdomen inoculation results in minimal mortality and lower bacterial loads than thorax inoculation. Inoculation into either site causes systemic infection. Differences in mortality and bacterial load are due to injury of the thorax and can be recapitulated by abdominal inoculation coupled with aseptic wounding of the thorax. This altered resistance appears to be independent of classical immune pathways and opens new avenues of research on the role of injury during defense against infection.

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Section: Discussionmentioning

confidence: 57%

Thorax Injury Lowers Resistance to Infection in Drosophila melanogaster

et al. 2014

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“…Brown, and YW) to the bacterial pathogens P. luminescens and E. faecalis and the nonpathogens E. coli and M. luteus. These bacteria are frequently used for understanding the insect immune system (23). Although the susceptibility of the five strains varied with the different types of infection, we consistently found that YW flies, and on one occasion w1118 flies, were particularly sensitive to all bacteria.…”

Section: Discussionmentioning

confidence: 65%

Nitric Oxide Levels Regulate the Immune Response of Drosophila melanogaster Reference Laboratory Strains to Bacterial Infections

Eleftherianos

Spivack

et al. 2014

Infect Immun

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Studies on the innate immune response against microbial infections in Drosophila melanogaster involve mutant strains and their reference strains that act as experimental controls. We used five standard D. melanogaster laboratory reference strains (Oregon R, w1118, Canton-S, Cinnabar Brown, and Yellow White [YW]) and investigated their response against two pathogenic bacteria (Photorhabdus luminescens and Enterococcus faecalis) and two nonpathogenic bacteria (Escherichia coli and Micrococcus luteus). We detected high sensitivity among YW flies to bacterial infections and increased bacterial growth compared to the other strains. We also found variation in the transcription of certain antimicrobial peptide genes among strains, with Oregon and YW infected flies showing the highest and lowest gene transcription levels in most cases. We show that Oregon and w1118 flies possess more circulating hemocytes and higher levels of phenoloxidase activity than the other strains upon infection with the nonpathogenic bacteria. We further observed reduced fat accumulation in YW flies infected with the pathogenic bacteria, which suggests a possible decline in physiological condition. Finally, we found that nitrite levels are significantly lower in infected and uninfected YW flies compared to w1118 flies and that nitric oxide synthase mutant flies in YW background are more susceptible to bacterial infection compared to mutants in w1118 background. Therefore, increased sensitivity of YW flies to bacterial infections can be partly attributed to lower levels of nitric oxide. Such studies will significantly contribute toward a better understanding of the genetic variation between D. melanogaster reference strains.

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“…The voucher specimen was deposited at Biofarmaka Laboratory, Faculty of Pharmacy, Hasanuddin University. Fly infection and assays for survival and bacterial growth The introduction of bacteria into the thorax of male adult flies, known as pricking, was carried out according to the established procedures 33 with modifications. Briefly, at 4-7 days after eclosion, the flies (10 flies per vial, 3 vials in each experiment) were pricked with bacterial suspension containing given numbers of bacteria (1 × 10 5 cfu/ml) per fly.…”

Section: Sample Preparationmentioning

confidence: 99%

“…This indicates that Toll immunodeficient flies were more sensitive to bacterial infection, supporting the reports of previous investigators. 33,35 Furthermore, treatment of the infected-immunodeficient flies with food containing ethanolic extract of Ulva reticulata at concentration of 25 mg/ml increased the survivorship of infected Toll mutant flies and reduced the bacterial load recovered from the corresponding mutant flies ( Figure 4A). Similar results were also observed in Draper mutant flies lacking for cellular innate immunity ( Figure 4B).…”

mentioning

confidence: 96%