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Imaging of Intracellular ATP in Organotypic Tissue Slices of the Mouse Brain using the FRET-based Sensor ATeam1.03YEMK
Published on: December 19, 2019
Enhancement of spontaneous synaptic activity in rat Purkinje neurones by ATP during development
Diana Casel1, Johannes Brockhaus, Joachim W Deitmer
1Abteilung Allgemeine Zoologie, Fachbereich Biologie, TU Kaiserslautern, Postfach 3049, D-67653 Kaiserslautern, Germany.
Abstract:
The establishment of functional synaptic connections and activity is a pivotal process in the development of neuronal networks. We have studied the synaptic activity in the developing rat cerebellum, and the contribution mediated by purinergic receptors. The mean frequency of the spontaneous postsynaptic currents (sPSCs) recorded with the whole-cell patch-clamp technique from Purkinje neurones in acute brain slices at room temperature, increased fourfold from 4.4+/-0.8 Hz at postnatal day 9/10 (n=23) to 17.8+/-1.6 Hz at postnatal day 17-20 (p17-p20; n=113; P<0.01). ATP, which increased the frequency of sPSCs by up to 100% (EC50=18 microM) in the third postnatal week, started to modulate the synaptic activity during the second postnatal week, which was determined by three processes: (1) the appearance of functional ATP receptors during p10-p12, (2) the enhancement of the sPSC frequency by endogenous ATP release becoming apparent after inhibition of ecto-ATPases by 6-N,N-diethyl-beta,gamma-dibromomethylene-D-adenosine-5-triphosphate (ARL67156; 50 microM) at p11-p12, and (3) with tonic stimulation of purinoceptors at p14, as revealed by the P2 receptor antagonist pyridoxal-phosphate-6-azophenyl-2',4'-disulphonic acid (PPADS, 10 microM). ATP had a similar effect at later stages (p24-p27) and at 35 degrees C. Our results suggest that endogenous release of ATP starts to enhance the synaptic activity in Purkinje neurones by the end of the second postnatal week.
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