J 2010

Fluorimetric determination of hydrogen peroxide production by the haemocytes of the wax moth Galleria mellonella L. (Lepidoptera: Pyralidae)

VAŠÍČEK, Ondřej; Ivana PAPEŽÍKOVÁ a Pavel HYRŠL

Základní údaje

Originální název

Fluorimetric determination of hydrogen peroxide production by the haemocytes of the wax moth Galleria mellonella L. (Lepidoptera: Pyralidae)

Název česky

Fluorimetric determination of hydrogen peroxide production by the haemocytes of the wax moth Galleria mellonella L. (Lepidoptera: Pyralidae)

Autoři

VAŠÍČEK, Ondřej; Ivana PAPEŽÍKOVÁ a Pavel HYRŠL

Vydání

European Journal of Entomology, 2010, 1210-5759

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

30105 Physiology

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 0.945

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14310/10:00045625

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova česky

Galleria mellonella; chemiluminescence; fluorescence; hydrogen peroxide; reactive oxygen species; Amplex Red; haemocytes; phagocytes; leukocytes; phagocytosis

Klíčová slova anglicky

Galleria mellonella; chemiluminescence; fluorescence; hydrogen peroxide; reactive oxygen species; Amplex Red; haemocytes; phagocytes; leukocytes; phagocytosis

Příznaky

Mezinárodní význam, Recenzováno
Změněno: 20. 5. 2011 12:21, doc. RNDr. Pavel Hyršl, Ph.D.

Anotace

V originále

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.

Česky

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.

Návaznosti

GP206/09/P470, projekt VaV
Název: Vliv inhibitorů biosyntézy eikosanoidů na imunitu zavíječe voskového Galleria mellonella
Investor: Grantová agentura ČR, Vliv inhibitorů biosyntézy eikonsanoidů na imunitu zavíječe voskového Galleria mellonella