Ca2+ signals during early lymphocyte activation in carp Cyprinus carpio L.
To measure cellular responses and the involvement of increased cytosolic Ca2 levels ([Ca2 ]i), peripheral blood leukocytes (PBL) of carp were loaded with the fluorescent intracellular Ca2 indicators Fluo-3 and Fura-2. Responses of lymphocytes to T-cell mitogen (phytohaemagglutinin, PHA), to B-cell mitogen (lipopolysaccharide, LPS) and to immunoglobulin (Ig) cross-linking with a monoclonal antibody to carp Ig were measured using flow cytometry. Both T-cell stimulation by PHA and B-cell stimulation by membrane Ig cross-linking evoked a rapid elevation of [Ca2 ]i. B-cell stimulation by LPS was not linked to an increase in [Ca2 ]i. As judged by the percentage of reacting cells, it was concluded that all Ig-positive lymphocytes reacted to Ig cross-linking by elevating [Ca2 ]i. At the single-cell level, the reactions of Fura-2-loaded cells were followed every 6 s using digital imaging microscopy. Both cells displaying spontaneous [Ca2 ]i oscillations and non-oscillating cells responded to stimulation with an increase in [Ca2 ]i, sometimes, in already oscillating cells, accompanied by an increase in frequency and/or amplitude of the oscillations. These results show that intracellular Ca2 responses of PBL upon activation resemble those in mammals and form a powerful tool for studies into cell-specific regulation.
| Main Authors: | , , , |
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| Format: | Article/Letter to editor biblioteca |
| Language: | English |
| Subjects: | B-cell cross-linking, Carp, Cyprinus carpio, Digital imaging microscopy, Fish, Flow cytometry, Fluo3, Fura-2, Intracellular Ca2+, Lipopolysaccharide, Lymphocytes, Phytohaemagglutinin, |
| Online Access: | https://research.wur.nl/en/publications/ca2-signals-during-early-lymphocyte-activation-in-carp-cyprinus-c |
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