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VOL. 76 (1), 3-22, 2010 CROSS-TALK BETWEEN GLUTAMATE AND NUCLEOTIDE...

Figure 5. Effect of L-Glu on CaMKII phosphorylation at Thr286/287 9 div
cerebellar granule cells. Neurons were stimulated with L-Glu 100 µM for 1 and
5 min, fixed with PFA and treated with antibodies that recognized either phospho-
CaM-Kinase II a/b (Thr286/287) or the total CaMKII (A). (B) Respectively, the
quantification of fluorescence obtained in cellular somas. Bars indicate the mean
± SEM of three independent experiments. Statistical significance was calculated by
anova test: ***, p < 0.001; *, p < 0.05. (C, D) Effect of L-Glu (100 µM) on ATP-
mediated responses measured in somas after pre-treatment with KN-62 10 µM (C)
and KN-93 10 µM (D). ATP-induced currents were studied 30 s and 5 min after
L-Glu application and were compared with the first ATP-response for each neuron.
Statistical significance was calculated by anova test: ***, p < 0.001; significantly
different from first ATP-mediated response; nd, not significantly different from
first ATP-mediated response. Scale bar: 30 µm.

We have used in this study cerebellar granule neurons which
express both glutamate and nucleotide receptors (2-4). With these
cells we measured the functional response by monitoring the agonist-
induced Ca2+ release in microfluorimetric experiments. Accordingly
with previous works, granule cells showed a heterogeneous pattern

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