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Zebrafish kidney phagocytes utilize macropinocytosis and Ca+-dependent endocytic mechanisms.
Abstract
BACKGROUND: The innate immune response constitutes the first line of defense against invading pathogens and consists of a variety of immune defense mechanisms including active endocytosis by macrophages and granulocytes. Endocytosis can be used as a reliable measure of selective and non-selective mechanisms of antigen uptake in the early phase of an immune response. Numerous assays have been developed to measure this response in a variety of mammalian and fish species. The small size of the zebrafish has prevented the large-scale collection of monocytes/macrophages and granulocytes for these endocytic assays. METHODOLOGY/PRINCIPAL FINDINGS: Pooled zebrafish kidney hematopoietic tissues were used as a source of phagocytic cells for flow-cytometry based endocytic assays. FITC-Dextran, Lucifer Yellow and FITC-Edwardsiella ictaluri were used to evaluate selective and non-selective mechanisms of uptake in zebrafish phagocytes. CONCLUSIONS/SIGNIFICANCE: Zebrafish kidney phagocytes characterized as monocytes/macrophages, neutrophils and lymphocytes utilize macropinocytosis and Ca(2+)-dependant endocytosis mechanisms of antigen uptake. These cells do not appear to utilize a mannose receptor. Heat-killed Edwardsiella ictaluri induces cytoskeletal interactions for internalization in zebrafish kidney monocytes/macrophages and granulocytes. The proposed method is easy to implement and should prove especially useful in immunological, toxicological and epidemiological research- text
- Animals
- Antigens
- Antigens: metabolism
- Calcium
- Calcium: metabolism
- Cell Surface
- Cell Surface: metabolism
- C-Type
- C-Type: metabolism
- Edwardsiella ictaluri
- Edwardsiella ictaluri: metabolism
- Endocytosis
- Endocytosis: physiology
- Flow Cytometry
- Fluorescein-5-isothiocyanate
- Kidney
- Kidney: cytology
- Lectins
- Mannose-Binding Lectins
- Mannose-Binding Lectins: metabolism
- Phagocytes
- Phagocytes: metabolism
- Pinocytosis
- Receptors
- Zebrafish
- Zebrafish: metabolism
- Veterinary Medicine