Measured increase in intracellular Ca(2+) during stimulated release of endogenous glutamate from human cerebrocortical synaptosomes.
Presynaptic terminals (synaptosomes) prepared from guinea pig and rat cerebral cortex release endogenous glutamate in a Ca(2+)-dependent manner in response to membrane depolarisation. In the present study, synaptosomes were prepared from human cerebral cortex removed in association with temporal lobe resections in epileptic patients. The cytosolic free Ca(2+) concentration increased from 474+/-66 before to 649+/-89 nM after 2 min depolarisation. The basal level of free cytosolic Ca(2+) is higher and the increase in response to depolarisation is more pronounced in human synaptosomes than observed in animal experiments. The Ca(2+)-dependent glutamate release, estimated as the difference between total - and the Ca(2+)-independent glutamate release, increased from 0 to 5.4+/-1.9 nmol/mg protein. The released amount of glutamate is larger than reported in animal models. These results demonstrate that membrane depolarisation of synaptosomes from human brain evokes a rapid rise in cytosolic free Ca(2+) and a more prolonged rise in synaptic, Ca(2+)-dependent glutamate release.
Vinje
M L
ML
Institute for Surgical Research, Rikshospitalet, The National Hospital, University of Oslo, N-0027, Oslo, Norway. mortenla@klinmed.uio.no
Valø
E T
ET
Røste
G K
GK
Berg-Johnsen
J
J
D015761:4-Aminopyridine / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002540:Cerebral Cortex / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D004833:Epilepsy, Temporal Lobe / Q000378:metabolism / Q000601:surgery; D018698:Glutamic Acid / Q000378:metabolism*; D006168:Guinea Pigs; D006801:Humans; D007700:Kinetics; D051381:Rats; D013574:Synaptosomes / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D013702:Temporal Lobe / Q000601:surgery
D018698:Glutamic Acid; D015761:4-Aminopyridine; D002118:Calcium