3H-D-aspartate release from cerebellar granule neurons is differentially regulated by glutamate- and K+-stimulation
✍ Scribed by Dr. B. Belhage; V. Rehder; G. H. Hansen; S. B. Kater; A. Schousboe
- Publisher
- John Wiley and Sons
- Year
- 1992
- Tongue
- English
- Weight
- 985 KB
- Volume
- 33
- Category
- Article
- ISSN
- 0360-4012
No coin nor oath required. For personal study only.
✦ Synopsis
Neurotransmitter release in response to either 55 mM K + or 25 pM glutamate as well as its dependency on Ca2' from different sources was compared in cultured glutamatergic cerebellar granule cells from rat brain. The intracellular Ca2+ concentration was monitored at the single cell level in neurites as well as cell bodies employing the fluorescent Ca" indicator fura-2. Transmitter release was assayed using 3H-Daspartate to label the exogenously accessable glutamate pools, which in these neurons is believed to also include the transmitter pool. In an attempt to distinguish whether transmitter release was dependent on an intact cytoskeleton or not, the colchicine-like drug Nocodazole, which also blocks transport of vesicles, was used. K+ -stimulated transmitter release consisted for the major part (around 70%) of a Ca2+dependent, Nocodazole sensitive release component and this K+-induced release appeared to be almost exclusively dependent on N-type Ca2' channels. In contrast, 50% of the glutamate-induced Ca2+-dependent release was triggered by Ca2' from a Dantrolene sensitive intracellular Ca2+ pool. Since these neurons undergo a pronounced maturational change in which neurotransmitter vesicles become increasingly prominent, the Cat' responses and transmitter release evoked by the two different stimuli were investigated as a function of the culture period. K + and glutamate were found to increase intracellular [Ca2+] differentially. In 1-day-old cultures K + elicited a small albeit significant increase in [Ca2 'Ii while glu-