Calcium uptake and calcium-dependent phosphorylation during development of rat brain neurons in culture

Neomi Zurgil, Nava Zisapel*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


The emergence of the capacities for calcium uptake and calcium-regulated protein phosphorylation during the development of embryonic brain neurons in tissue culture was examined. In the maturing cells, the enhancement in 45Ca2+-uptake upon stimulation with high K+ increased by 3-4 fold during the second week in vitro, in parallel to an increase in the capacity for high K+-induced Ca2+-dependent release of prelabeled [3H]dopamine. The pattern of incorporation of [32Pi]phosphate into the major phosphoproteins in maturing cells under nonstimulating conditions also changed during cell development: the incorporation of 32Pi into two proteins of apparent molecular weights - 55,000 and 43,000 dalton - increased, but decreased in a 45,000 dalton protein. Stimulation of mature cells (after 10-11 days in vitro) resulted in a Ca2+-dependent increase in the amount of 32Pi incorporated into the 43,000 dalton protein and a decrease in the amount incorporated into the 55,000 dalton protein. This calcium-regulated phosphorylation pattern was not observed until 6 days in vitro. Introduction of Ca2+ into the immature cells by means of the Ca2+ ionophore A23187 did not alter the phosphorylation pattern and did not cause neurotransmitter release. The amount of [35S]methionine incorporated into a 43,000 dalton protein which comigrated with the 43,000 dalton phosphoprotein also increased upon cell maturation. The results suggest that this phosphoprotein (which does not comigrate with nonphosphorylated actin on two-dimensional polyacrylamide gels) develops in the cells in parallel to the emerging processes of the stimulation-induced calcium entry and calcium-dependent neurosecretion.

Original languageEnglish
Pages (from-to)293-303
Number of pages11
JournalDevelopmental Brain Research
Issue number2
StatePublished - Apr 1984


  • brain neurons
  • calcium
  • development
  • phosphorylation


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