Express Method for Determination of Low Value of Trans-membrane Potential of Living Cells with Fluorescence Probe: Application on Haemocytes at Immune Responses

Glazachev, Yuriy I.; Semenova, Alexandra; Kryukova, Natalia A.; Slepneva, Irina A.; Глупов, В. В.

doi:10.1007/s10895-012-1062-0

Cited by 6 publications

(3 citation statements)

References 13 publications

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“…The method to measure the transmembrane potential of cell membrane was evaluated for the case of low potential value using the fluorescence probe 4‐(4‐dimethylaminostyryl)‐1‐methylpyridinium, DSM (Glazachev et al., ), kindly provided by Dr. N. Kolosova (Institute of Cytology & Genetics of RAS SB). The DSM stock solution in water was about 1 mM, which is its ultimate solubility.…”

Section: Methodsmentioning

confidence: 99%

VENOM FROM THE ECTOPARASITIC WASP Habrobracon hebetor ACTIVATES CALCIUM‐DEPENDENT DEGRADATION OF Galleria mellonella LARVAL HEMOCYTES

Kryukova

Chertkova

Semenova

et al. 2015

Arch Insect Biochem Physiol

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Ectoparasitoids inject venom into hemolymph during oviposition. We determined the influence of envenomation by the parasitoid, Habrobracon hebetor, on the hemocytes of its larval host, Galleria mellonella. An increase in both intracellular Са(2+) content and phospholipase C activity of the host hemocytes was recorded during 2 days following envenomation by the parasitoid. The decreased hemocyte viability was detected 1, 2, and 24 h after the envenomation. Injecting of the crude venom (final protein concentration 3 μg/ml) into the G. mellonella larvae led to the reduced hemocyte adhesion. The larval envenomation caused a decrease in transmembrane potential of the hemocytes. These findings document the suppression of hemocytic immune effectors in the parasitized host larvae.

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Section: Methodsmentioning

confidence: 99%

VENOM FROM THE ECTOPARASITIC WASP Habrobracon hebetor ACTIVATES CALCIUM‐DEPENDENT DEGRADATION OF Galleria mellonella LARVAL HEMOCYTES

Kryukova

Chertkova

Semenova

et al. 2015

Arch Insect Biochem Physiol

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“…Intriguingly, little examination of the relationship between hemocyte function and membrane potential, as well as with endogenous and exogenous electrical fields, has been performed in insects. Caterpillar hemocytes undergo depolarization during immune stimulation [ 42 ], although the mechanism(s) and significance of this phenomenon are unknown. Drosophila melanogaster hemocyte calcium transients are required for orientation and migration, and disruption by a parasitoid venom component reduces cellular immunity [ 43 ].…”

Section: Discussionmentioning

confidence: 99%

Polydnavirus Innexins Disrupt Host Cellular Encapsulation and Larval Maturation

Zhang

Turnbull

2021

Viruses

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Polydnaviruses are dsDNA viruses associated with endoparasitoid wasps. Delivery of the virus during parasitization of a caterpillar and subsequent virus gene expression is required for production of an amenable environment for parasitoid offspring development. Consequently, understanding of Polydnavirus gene function provides insight into mechanisms of host susceptibility and parasitoid wasp host range. Polydnavirus genes predominantly are arranged in multimember gene families, one of which is the vinnexins, which are virus homologues of insect gap junction genes, the innexins. Previous studies of Campoletis sonorensis Ichnovirus Vinnexins using various heterologous systems have suggested the four encoded members may provide different functionality in the infected caterpillar host. Here, we expressed two of the members, vnxG and vnxQ2, using recombinant baculoviruses in susceptible host, the caterpillar Heliothis virescens. Following intrahemocoelic injections, we observed that >90% of hemocytes (blood cells) were infected, producing recombinant protein. Larvae infected with a vinnexin-recombinant baculovirus exhibited significantly reduced molting rates relative to larvae infected with a control recombinant baculovirus and mock-infected larvae. Similarly, larvae infected with vinnexin-recombinant baculoviruses were less likely to survive relative to controls and showed reduced ability to encapsulate chromatography beads in an immune assay. In most assays, the VnxG protein was associated with more severe pathology than VnxQ2. Our findings support a role for Vinnexins in CsIV and more broadly Ichnovirus pathology in infected lepidopteran hosts, particularly in disrupting multicellular developmental and immune physiology.

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“…Intriguingly, little examination of the relationship between hemocyte function and membrane potential, as well as with endogenous and exogenous electrical fields, has been performed in insects. Caterpillar hemocytes undergo depolarization during immune stimulation (43), although the mechanism(s) and consequences of this are unknown. Additionally, disruption of the SERCA pump by venom components in hemocytes of Leptoplina -parasitized D. melanogaster larvae inhibits host immunity (44).…”

Section: Discussionmentioning

confidence: 99%

Ectopic Expression of Virus Innexins inHeliothis virescensDisrupts Hemocyte-Mediated Encapsulation and Host Viability

Zhang

Turnbull

2019

Preprint

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1.AbstractPolydnaviruses are dsDNA viruses associated with endoparasitoid wasps. Delivery of the virus during parasitization of a caterpillar and subsequent virus gene expression is required for production of an amenable environment for parasitoid offspring development. Consequently, understanding of Polydnavirus gene function provides insight into mechanisms of host susceptibility and parasitoid wasp host range. Polydnavirus genes predominantly are arranged in multimember gene families, one of which is the vinnexins, which are virus homologues of insect gap junction genes, the innexins. Previous studies of Campoletis sonorensis Ichnovirus Vinnexins using various heterologous systems have suggested the four encoded members may provide different functionality in the infected caterpillar host. Here, we expressed two of the members, vnxG and vnxQ2, using recombinant baculoviruses in susceptible host, the caterpillar Heliothis virescens. Following intrahemocoelic injections, we observed >90% of hemocytes (blood cells) were infected, producing recombinant protein. Larvae infected with a vinnexin-recombinant baculovirus exhibited significantly reduced molting rates relative to larvae infected with a control recombinant baculovirus and mock infected larvae. Similarly, larvae infected with vinnexin-recombinant baculoviruses were less likely to molt relative to controls, and showed reduced ability to encapsulate chromatography beads in an immune assay. In most assays, the VnxG protein was associated with more severe pathology than VnxQ2. These results, in light of previous findings, support that Polydnavirus Vinnexin gene family members may provide complementary, rather than redundant, effects. This in turn indicates a need to test gene family member functionality across infected hosts for effects to determine member contribution to host range.2.ImportancePolydnaviruses are obligate mutualistic associates of highly speciose wasp taxa that parasitize caterpillars. Expression of Polydnavirus-encoded genes in hosts parasitized by wasps is necessary for successful parasitization, and an unusual genome structure including multiple-membered gene families is hypothesized to contribute to host manipulation. We have tested this hypothesis by in vivo expression of two members of a family of Polydnavirus homologues of Innexins, or insect gap junction proteins. Previous findings demonstrated that the two Vinnexins induce different physiological alterations in heterologous systems. Here, in host caterpillars, we observed differential alteration by the two proteins of host immune cell (hemocyte) bioelectrical physiology and the immune response of encapsulation. Not only do our data suggest a linkage between cellular bioelectricity and immunity in insects, but they support that gene family expansion has functional consequences to both Polydnavirus and host wasp success.

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Express Method for Determination of Low Value of Trans-membrane Potential of Living Cells with Fluorescence Probe: Application on Haemocytes at Immune Responses

Cited by 6 publications

References 13 publications

VENOM FROM THE ECTOPARASITIC WASP Habrobracon hebetor ACTIVATES CALCIUM‐DEPENDENT DEGRADATION OF Galleria mellonella LARVAL HEMOCYTES

VENOM FROM THE ECTOPARASITIC WASP Habrobracon hebetor ACTIVATES CALCIUM‐DEPENDENT DEGRADATION OF Galleria mellonella LARVAL HEMOCYTES

Polydnavirus Innexins Disrupt Host Cellular Encapsulation and Larval Maturation

Ectopic Expression of Virus Innexins inHeliothis virescensDisrupts Hemocyte-Mediated Encapsulation and Host Viability

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