Ca2+ signals during early lymphocyte activation in carp Cyprinus carpio L.

B. M Lidy Verburg-Van Kemenade, Jeroen Saeij, Gert Flik, Peter H G M Willems

Research output: Contribution to journalArticle

18 Scopus citations

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 languageEnglish (US)
Pages (from-to)591-598
Number of pages8
JournalJournal of Experimental Biology
Volume201
Issue number4
StatePublished - Feb 1998
Externally publishedYes

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)

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  • Cite this

    Verburg-Van Kemenade, B. M. L., Saeij, J., Flik, G., & Willems, P. H. G. M. (1998). Ca2+ signals during early lymphocyte activation in carp Cyprinus carpio L. Journal of Experimental Biology, 201(4), 591-598.