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Purification of Biotinylated Cell Surface Proteins from Rhipicephalus microplus Epithelial Gut Cells
Published on: July 23, 2017
An insight into the functional role of antioxidant and detoxification enzymes in adult Rhipicephalus microplus female
Gabriela A Sabadin1, Tiago B Salomon2, Milane S Leite3
1Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Avenida Bento Gonçalves, 9500 Porto Alegre, RS, Brazil.
Abstract:
Ticks have developed physiological adaptations to transport, store, metabolize and secrete toxic components from the diet and environment. Different classes of enzymes are involved in these processes, however, the role of several of them is not yet characterized in Rhipicephalus microplus. In this context, this work investigated the action of antioxidant and detoxification enzymes, as well as the levels of essential cellular reductants in R. microplus partially engorged females (PEF) and fully engorged females (FEF). Results demonstrated that enzymes transcriptional levels and enzymatic activity from ovary and fat body were higher in PEF than in FEF, except for ovary Glutathione peroxidase (GPx), which was the only enzyme showing highest activity in the FEF stage. These results indicated a higher demand for antioxidant potential in these organs at the initial feeding phase than during egg-laying. In midgut, however, there was more variability in the transcriptional levels and activity of the different enzymes between the PEF and FEF phases. Similar NADPH levels were found in PEF and FEF phases, suggesting a remarkable capacity to maintain a regular supply of reducing power, despite the developmental changes and large intake of heme and iron. However, reduced glutathione (GSH) levels were variable between PEF and FEF when distinct organs were compared. Taken together, our data suggest a higher demand for reducing potential in FEF ticks. The silencing of catalase (CAT) or thioredoxin reductase (TRx) genes in females did not impair feeding, egg-laying capacity, or larvae hatching. CAT-silenced ticks had increased ovary peroxidase activity, a possible compensatory antioxidant mechanism. Altogether, the results shed light on the complexity of the antioxidant and detoxification enzyme system in ticks and its involvement in different physiological mechanisms.
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