Effect of MTA under OGD and AMPA excitotoxicity.

<p>Panels a and b: the effect of APV (100 µM) or MTA in OGD conditions in rat mixed cultures of neurons and astrocytes. Cell death was expressed as the LDH activity at the beginning of any treatment: *<i>p</i><0.05 compared to IAA-treated cells; # <i>p</i><0.05 compared to control; ** <i>p</i><0.01 compared to IAA-treated cells; ## <i>p</i><0.01 compared to control. <b>c</b>) primary oligodendrocytes derived from rat optic nerve subjected to AMPA excitotoxicity (10 µM and 100 µM) in which cell death was measured with calcein-AM, as indicated in Methods. *p<0.05 compared with cells treated with AMPA (Unpaired t-test). Values represent the average ± SEM and were obtained from at least three independent experiments performed in duplicates.</p

Effect of MTA and A_2AR antagonists in an acute mouse model of PD.

<p>The effect of MTA (30 mg/kg), MSX-3 (9 mg/kg) or both in combination on the survival of dopaminergic neurons (TH⁺ cells) within the <i>substantia nigra</i> of the brain of mice treated with MPTP. a) Representative images of the neurodegeneration of dopaminergic cells in the different groups. b) Stereological counts of TH⁺ neurons in control and drug-treated animals (n = 6 per group). The data represent the mean±SEM: *<i>p</i><0.05 comparing with the MPTP group; # <i>p</i><0.05 comparing with the control group (One-way ANOVA, LSD test as a post-hoc). Scale bar: 50 µm.</p

MTA effects in chronic pilocarpine-induced status epilepticus (SE).

<p>a–f: Representative images of Neu N immunoreactivity in the hippocampus are shown 3 days after sham treatment (a) or pilocarpine-induced SE (b, c), or 30 days after SE (d–f). MTA (30 mg/kg) was administered pre-SE (d) or post-SE (c,f) induction. Cell loss is already apparent by 3 days (arrows in b) and is marked by 30 days (e) after SE, but it appears attenuated in MTA-treated animals at both timepoints (c,d,f). g–h) Bar plots show quantification of NeuN-positive cells in CA3(g) or CA1(h) 30 days after SE in animals pre-treated with MTA. The data represent the mean ± SEM. *p<0.05; **p<0.001; ANOVA with Tukey HSD post –hoc test.</p

Effective permeability (Pe) values in the PAMPA assay and in the <i>in vitro</i> model of BBB.

<p>*<i>transport confirmed by HPLC-MS.</i></p

Effect of MTA on brain ischemia.

<p>(a) Representative TTC stained sections of vehicle and MTA treated animals (30 mg/kg/twice daily, i.p.) 3 days after the induction of transient focal ischemia. Histogram (right) showing the infarct volume calculated from TTC stained slices in vehicle- and MTA-treated rats (n = 5 in each group). b) Representative microphotographs of Fluoro Jade C staining after rat transient forebrain ischemia (n = 5 in the vehicle group; n = 6 in the MTA-treated group). MTA (30 mg/kg) was administered 30 min after triggering ischemia. Quantification of Fluoro Jade positive cells per mm length of CA1 pyramidal layer (right). The data represents the mean ± SEM: **p<0.01 compared to the vehicle (Student t-test). Scale bar 100 µm.</p

Effect of MTA under NMDA excitotoxicity.

<p>Effect of MTA co-treatment (a) or MTA pre-treatment (b) on NMDA-induced caspase 3 activation in rat pure primary neuronal cultures. Effect of MTA co-treatment (c) or MTA pre-treatment (d) on NMDA-induced caspase 3 activation in rat mixed astrocyte-neuron cultures. Caspase 3 activity (units of fluorescence per milligram of protein per hour) was determined in cells treated with 300 µM NMDA, in the presence or absence of MTA (panels a and b: 100, 250 and 500 µM; panels c and d: 250 µM) or 10 µM MK-801 (MK; NMDA receptor antagonist). The results are expressed as the mean ± SEM of at least four independent experiments performed in triplicate: *p<0.05, **p<0.01***<i>p</i> <0.001 compared with cells treated with NMDA; # <i>p</i><0.05, ## <i>p</i><0.01, ### <i>p</i><0.001 compared with vehicle cells. One-way analysis of variance (ANOVA) and Bonferroni's t-test for multiple comparisons.</p