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Development of calcium action potentials in mouse hippocampal cell cultures


Developmental Brain Research 8(1): 39-52
Development of calcium action potentials in mouse hippocampal cell cultures
Action potentials (AP) mediated by Ca ions were recorded intracellularly from neurons in cell cultures prepared from fetal mouse hippocampus and examined between 18 h-46 days in culture. These AP occurred in media containing Na (4-14 mM), Ca ions (5-8 mM) and tetrodotoxin (1 .mu.g/ml), were enhanced by tetraethylammonium [TEA] ions (5-20 mM), and were blocked by Co ions (10 mM). In cultures prepared from 18-day fetuses, the incidence of neurons with Ca AP activity was 46% (n = 35) from 18 h to 2 days. For the next 6 wk in culture the incidence was between 83-89%. The lower incidence of Ca AP is due to temporary loss of Ca AP capability immediately following culture preparation, rather than a lack of development of this property by fetal day 18 because cultures of neurons chronologically equivalent to fetal ages 16-18 days have a high incidence of Ca AP (91%, n = 22). Na-dependent AP can be demonstrated in cells which have Ca AP by perfusion techniques. These dual AP mechanisms persist within the same cell throughout the time span of the cultures in this study. Special features of Ca AP in cultured hippocampal neurons included the following: Ca AP were optimally generated when the membrane potential was depolarized to -40 mV except with TEA ions in the medium. Uner this condition, optimal activity occurred between -60 and -70 mV; AP morphology suggested the presence of multiple generator sites and the distribution of Ca channels over both soma and processes; hyperpolarizing afterpotentials were well developed in some, but not all cells with Ca AP; up to 25% of the cells in selected cultures exhibited rhythmic pacemaker activity with a frequency which increased linearly from about 1-7 Hz with increasing steady current injection; Mn ions did not effectively block Ca AP in cultured hippocampal neurons and, in fact, appeared to support AP activity. Marked hyperpolarizing afterpotentials followed some Mn AP.

Accession: 005132817

DOI: 10.1016/0165-3806(83)90155-4

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