Abstract
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.
Original language | English (US) |
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Pages (from-to) | 591-598 |
Number of pages | 8 |
Journal | Journal of Experimental Biology |
Volume | 201 |
Issue number | 4 |
State | Published - Feb 1998 |
Externally published | Yes |
Keywords
- B-cell cross-linking
- Carp
- Cyprinus carpio
- Digital imaging microscopy
- Fish
- Flow cytometry
- Fluo3
- Fura-2
- Intracellular Ca
- Lipopolysaccharide
- Lymphocytes
- Phytohaemagglutinin
ASJC Scopus subject areas
- Agricultural and Biological Sciences(all)
- Agricultural and Biological Sciences (miscellaneous)