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Experiments were performed at room temperature (22�C25��C). Slices were superfused with extracellular solution composed of: 125 mm NaCl, 2.5 mm KCl, 1.25 mm NaH2PO4, 25 mm NaHCO3, 4 mm CaCl2, 4 mm MgCl2, 11 mm glucose and equilibrated with 95% O2�C5% CO2. Bicuculline (10 ��m) was added to the bath to inhibit GABAA receptors. Intracellular solution was composed http://www.selleckchem.com/products/AZD0530.html of: 122 mm cesium methanesulfonate, 2 mm NaCl, 10 mm Hepes, 10 mm EGTA, 2 mm MgCl2, 4 mm ATP-Na (pH 7.3). A glass microelectrode was placed in the hilus of the dentate gyrus to stimulate MFs. While recording from a pyramidal cell, the stimulating electrode was moved to a position where a sharp EPSC with fixed latency was evoked. Stimulation intensity (200 ��s pulse; http://en.wikipedia.org/wiki/Diglyceride of their immature state. In that case, the group II mGluR agonist LCCG-1 (10 ��m) was applied to confirm that MFs were stimulated. LCCG-1 was also occasionally applied even when the facilitation ratio was above 3, and, in such cases, it always markedly inhibited EPSCs (Supplemental Fig. 1). Recordings were analysed using IGOR pro (WaveMetrics, Inc., Lake Oswego, OR, USA). Low-frequency facilitation was evoked by stimulating MFs at frequencies of 0.1 and 1 Hz. For Sr2+ experiments, slices were bathed in an extracellular solution containing SrCl2 (2 mm) and a low concentration of CaCl2 (0.03 mm). Asynchronous EPSCs (aEPSCs) were evoked by trains of five stimuli at 50 Hz repeated http://www.selleckchem.com/products/GDC-0941.html 100 times at 30 s intervals. Only events with amplitudes larger than 7.5 pA and occurring within a 400 ms time window beginning 50 ms after each train were considered for statistical analysis. Unless otherwise mentioned, values are expressed as means ��s.e.m. ANOVA followed by Bonferroni's post hoc test, the Mann�CWhitney U test or the Kolmogorov�CSmirnov (KS) test were used where appropriate. All drugs were obtained from Tocris Bioscience (Bristol, UK) or Ascent Scientific (Avonmouth, UK). Mice at P7, P14 and P21 (5 animals of each genotype and age) were deeply anaesthetized using a mixture of Ketavet (Parke-Davis) and Domitor (Pfizer) and perfused transcardially with 0.9% NaCl for 1 min followed by 4% formaldehyde in 0.1 m phosphate buffer (PB) for 10 min. Brains were removed, post-fixed in the same fixative for 20 h at 6��C, freeze-protected with 1 m sucrose in 0.1 m PB and frozen at ?40��C in isopentane.