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Intracellular free calcium level and its response to cAMP stimulation in developing Dictyostelium cells transformed with jellyfish apoaequorin cDNA
Author(s) -
Saran Shweta,
Nakao Hajime,
Tasaka Masao,
Iida Hidetoshi,
Tsuji Frederick I.,
Nanjundiah Vidyanand,
Takeuchi Ikuo
Publication year - 1994
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(94)80626-8
Subject(s) - aequorin , dictyostelium discoideum , aequorea victoria , jellyfish , biology , intracellular , calcium , microbiology and biotechnology , stimulation , second messenger system , biophysics , biochemistry , chemistry , green fluorescent protein , fishery , organic chemistry , gene , neuroscience
A new method is described for measuring intracellular free calcium concentrations, [(Ca 2+ i )], in the cells of Dictyostelium discoideum transformed with apoaequorin cDNA of the jellyfish, Aequorea victoria . Aequorin, a calcium‐specific indicator, was regenerated in vivo from apoaequorin produced in the cells by incubation with coelenterazine. The results showed that [(Ca 2+ ) i ] in developing cells markedly increases at the aggregation stage and again at the culmination stage after a temporary drop at the migration stage. Except for the vegetative stage, the cells at all stages of development exhibit a sharp transient increase in [(Ca 2+ ) i ] upon stimulation with a cAMP (50 nM) pulse, high responses being observed at the migration and culmination stages. Separated prestalk cells of migrating slugs contain more than twice as much [(Ca 2+ ) i ] and show three times as large a response to cAMP stimulation as prespore cells.