Fluorimetric determination of hydrogen peroxide production by the haemocytes of the wax moth Galleria mellonella L. (Lepidoptera: Pyralidae)
| Authors | |
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| Year of publication | 2010 |
| Type | Article in Periodical |
| Magazine / Source | European Journal of Entomology |
| MU Faculty or unit | |
| Citation | |
| Field | Physiology |
| Keywords | Galleria mellonella; chemiluminescence; fluorescence; hydrogen peroxide; reactive oxygen species; Amplex Red; haemocytes; phagocytes; leukocytes; phagocytosis |
| Description | The aim of this work was to investigate whether haemocytes of Galleria mellonella (Lepidoptera: Pyralidae) larvae can produce reactive oxygen species (ROS) in comparison to human blood phagocytes. The production of ROS was measured first with the use of luminol - enhanced chemiluminescence of unstimulated and stimulated (four activators with different mode of action) haemolymph or isolated haemocytes. However, spontaneous as well as the activated ROS generation remained at the background level. In subsequent experiments an ultrasensitive fluorescence method using Amplex Red reagent to detect hydrogen peroxide (H2O2) was tested. After optimization, Amplex Red was successfully used for determination of H2O2 production in both unstimulated and stimulated haemocytes. To determine the role of pH and ions during measurement, several diluent solutions were tested. We have shown that Ca2+ and Mg2+ ions are less important for the reaction in insects as compared to mammalian cells. |
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