1.0. FSL NMDAR-fEPSP 0.8. amplitude (mv) Intensity (µa) 2.0 SD FSL Time (ms)

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1 a AMPAR-fEPSP slope (mv/ms) NMDAR-fEPSP amplitude (mv) Intensity (µa) Intensity (µa) b 2. PPF Ratio (fepsp2/fepsp1) Time (ms) Supplementary Figure 1. Evoked glutamatergic transmission and PPF is normal in rats. a, Input/output (I/O) curves obtained in CA1 area following collateral stimulation in hippocampal slices from control () and adult rats using a range of stimulus intensities (1-7 µa) in the absence (Left) and presence (Right) of 1 μμ DNQX to record evoked AMPAR and NMDAR-fEPSP, respectively. In both cases I/O curves from and rats followed a sigmoidal function (AMPAR-fEPSP slope: r2=.67 and.79, and respectively; NMDAR-fEPSP: r2:.66 and.59, and respectively) of the stimulus intensity and no statistical differences were observed between groups (P >.5). b, Paired-pulse facilitation ratio (second fepsp slope/first fepsp slope) evoked at different interstimulus intervals (from 5 to 5 ms) in hippocampal slices from rats is statistically indistinguishable from that evoked in WT mice.

2 15 4 sipscs Frequency (Hz) 1 5 sipscs Amplitude (pa) Supplementary Figure 2. Spontaneous IPSCs in the hippocampus of and rats. Quantification of the frequency ( vs. (Hz): 7.5 ± 1.4 vs. 8.7 ± 1.8) and amplitude (pa: 23.5 ± 6.9 vs ± 3) of the sipscs shows similar mean values between groups (P =.56 and P =.7, Mann-Whitney test; N = 4-5 cells, 4 animals per group).

3 fepsp slope Time (min) Supplementary Figure 3. Normal medial-perforant-path-dentate gyrus LTP in rats. Three trains of HFS stimulation (2 trains, each of eight pulses at 2 Hz with an interval of 1 min; black arrows) in the perforant pathway of adult rats induce a strong MPP-DG LTP. The extent of potentiation is similair to what we have observed in rats (data not shown). Data is presented as means ± s.e.m (n = 5 slices, 4 animals). Black bar: bath application of picrotoxin (5μM).

4 a fepsp slope _control _D-serine b fepsp slope c Time (min). _control _D-serine fepsp slope (mv/ms) d _control _D-serine Intensity (µa) fepsp slope (mv/ms) DNQX DNQX + D-Serine DNQX + D-Serine + AP5 _control _D-serine Intensity (µa) D-Serine effect on NMDAR-fEPSP Supplementary Figure 4. Effect of D-serine application in CA1-LTP in control rats and in evoked glutamatergic transmission in and control rats. a, Three HFS (1 Hz, 1s, 1 s apart) application in the SC pathway after 1 µm D-serine bath application (green bar). indicates HFS onset. b, Summary of experiments in a. At 4-45 min after the induction, potentiation in the _D-serine group was 156 ± 2 % versus 147 ± 12 % in the _control group (n = 5-6 slices, 4-5 animals per group). c, Input/output curves obtained in CA1 area following collateral stimulation in hippocampal slices from control (; left) and (right) adult rats using a range of stimulus intensities (1-7 µa) before and after 1 min of 1 µm of D-serine application. In all groups, fepsp slope is a sigmoidal function (r2 =.66 and.73, _control and _D-serine respectively and r2 =.93 and.93, _control and _D-serine, respectively) of the stimulus intensity. d, (Left) representative traces of evoked NMDAR-fEPSP recorded from hippocampus slices of and rats in the presence of 1 μμ DNQX ( ), + 1 μμ D-Serine ( ) and + 25 μμ AP5 ( ). (Right) Quantification of the effect of 1 μμ D-serine application on the amplitude of evoked NMDAR-fEPSP response in and measured as the fepsp amplitude in the presence of DNQX + D-serine divided by the fepsp amplitude in DNQX. No differences between groups were obseved (P =.76; n = 6-8 slices, 3-4 animals per group). In a-d, data represent means ± s.e.m.

5 a 8 b 4 n.s Exploration time (s) Immobility time (% control) * c A B Saline A B D-serine Saline Saline D-serine Exploration time (s) A B A B A B Saline Saline D-serine Recognition Index * ** Saline Saline D-serine Supplementary Figure 5. Effect of acute D-serine administration on the NOR and FST tests in rats. a, Acute systemic administration of D-serine (6mg/kg, s.c.) does not restore the motivation deficit in rats. During the learning phase of the NOR, both saline and D-serine rats explore same total amount of time (56.7 ± 3.5 s ( saline) vs. 6.9 ± 4.6 s ( D-serine). The rats showed no preferences for any object: object A/ object B ratio:.9 ±.1 ( saline) and 1.2 ±.1 ( D-serine). b, Effect of acute D-serine treatment in the FST. Relative to control ( saline), saline rats were more immobile over a 5 min FST (*). D-serine rats did not show any statistical difference compared to the saline group (P >.5). c, Effect of acute D-serine injection (6 mg/kg) on memory recognition. The exploration time for rats were matched to the time rats actively explored during the 15 minutes exploration session (left). D-serine injected rats showed increased memory recognition when compared to saline group quantified as the recognition index (right). In a-d, data represent means ± s.e.m. In b-c, * P <.5; ** P<.1, One-way ANOVA followed by Bonferroni s test; n = 6-8 animals per group.

6 a acsf D-serine 1 s 1 pa % Frequency Time (min) % Amplitude Time (min) Supplementary Figure 6. Effect of D-serine application in spontaneous glutamatergic transmission in rats. a, (Left) representative traces of sepscs recorded from hippocampus slices of rats before (baseline), and during application of 1µM D-serine. (Right) quantification of the effect of D-serine application confirms no effect over the frequency (left) and amplitude (right) of spontaneous EPSCs. Data represent means ± s.e.m.

7 b a MoDG GrDG n.s 6 Astrocytes body area (μm2) PoDG 4 2 PoDG GrDG MoDG Supplementary Figure 7. Hippocampal astrocytic morphology in the dentate gyruds of and rats. a, (Left panels) representative images of DG granular, molecular and polymorphic layers of control () and hippocampal slices that were immunostained against GFAP (scale bar in black: 1 µm). (Right panels) Higher magnification of the boxed areas in the left panels (scale bar in black: 3 µm). b, Quantification of the astrocytic body area shows no statistical difference between groups, P =.11, n.s (not significant), Mann-Whitney test, N = 12 cells, 3 animals per group). Data in b represent the mean ± s.e.m. GrDG, granular cell layer; MoDG, molecular cell layer, PoDG, polymorphic layer, DG, dentate gyrus.

8 Strain (+ treatment) Rise (ms) Decay (ms) (baseline) 2.6 ± ±.3 (+ DL-TBOA) 2.5 ± ± ±.1 6. ± ± ±.3 Supplementary Table 1. Rise and decay times of spontaneous EPSCs recorded in the soma of CA1 pyramidal cells in control () and rats. 5 min of DL- TBOA (5 M) bath application does not affect the kinetics of sepscs. Values of different cell recordings from hippocampal slices of and rats performed during different experiments at normal conditions (regular acsf) are also showed to note the lack of differences between groups.

9 Supplementary methods: Enantioselective LC fluorescence detection for the quantification of D-serine in brain homogenates after derivatisation with o-phthalaldehyde The stock solution of D-serine (2 mm) was prepared in.1m HCl. Further dilutions (4; 2, 4; 8; 2 µm) were made in.1m HClO 4. Calibration standards are prepared by the standard addition method: 15 µl homogenate is mixed with 15 µl of standard solution, whereafter this mixture is neutralised and derivatised. The derivatisation agent was prepared by dissolving 1 mg o-phthalaldehyde and 2 mg N-isobutyryl-L-cysteine in 1 µl methanol and 9 µl.2m borate buffer ph 9.5. Each standard/sample was neutralised with an equal amount of NaOH.1M and 9µL of neutralised standard/sample was derivatised with 9µL of the o-phtalaldehyde/ N- isobutyryl-l-cysteine mixture. LC experiments were performed using a Shiseido CAPCELL PAK MG (25 cm x 2. mm i.d., 5 µm particles) C 18 column (ph stability 2 1, Analis, Namur, Belgium). Compounds were eluted by gradient elution with a mobile phase delivered at.17 ml/min using a Dionex pump (model P58A HPG, the Netherlands). An external vacuum degasser (Bioanalytical Systems, West Lafayette, USA) was used. Injections (centered loop 2 µl) were made using a refrigerated Gilson autosampler 231 XL (Gilson,Villiers Le Bel, France). The rinsing solution consisted of 1% acetonitrile and 9% water which was purified with a Arium Pro UV-DI purification system (Sartorius Stedim Biotech, Goettingen, Germany). Gradient elution was performed using mobile phase A (.25M phosphate solution ph 9) and mobile phase B (methanol/water 6:4). The linear LC gradient was 3-37% B in 5 min, 85% B in.1 min, 85-95% B in 1 min, 95% B for 5 min and 95 3% B in.1 min. The total run time was 8 min. For fluorescence detection, a Shimadzu spectrofluorometric

10 detector RF-1Axl was modified by introducing a 2 µl semi-microcell (Shimadzu, Duisburg, Germany). Derivates were measured at excitation and emission wavelengths of 34 and 45 nm, respectively. Integration of the chromatograms was performed by the integration computer program Clarity (DataApex, Antec, Zoeterwoude, the Netherlands).

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