Research report
Effect of potassium depolarization on phosphate-activated glutaminase activity in primary cultures of cerebellar granule neurons and astroglial cells during development

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Abstract

The cerebellar granule cells are believed to be glutamatergic neurons. During the normal development of granule cells grown in a chemically defined medium, the specific activity of phosphate-activated glutaminase increased from 60 at 3 days to 150 (nmol/h/mg protein) at 15 days in vitro. Treatment with 25 mM K+ for the last 2 days elevated glutaminase activity in an age-dependent manner: about 100% at 3 and 6 days, 75% at 10 days, and 40% at 15 days in vitro. The enhancement of glutaminase in granule cells was dose-dependent. The half-maximal effect was obtained at about 20 mM K+, whereas the maximum concentration, which produced about a 2.5-fold increase in 3-day-old cultures was about 40 mM K+. The voltage-sensitive Na+ channel inhibitor tetrodotoxin had no effect on the depolarization-induced activity in granule cells. However, the increase in glutaminase by 25 mM K+ was significantly blocked by both organic (nifedipine) and inorganic (Ni2+ and Mg2+) calcium antagonists, indicating that elevation in activity may be mediated through transmembrane Ca2+ entry into granule cells. In contrast to neurons, in cultured cerebellar astrocytes, the activity of glutaminase slightly decreased during development, and treatment with 25 mM K+ had no significant effect on this enzyme activity. The present findings, together with previous observations, would indicate that depolarization with K+, which is believed to mimic in vivo presynaptic stimulation, could be one of the mechanisms that selectively controls the development and function of neurons, when measured in terms of the activity of the enzymes involved in the synthesis of cell-specific neurotransmitters.

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