Neonatal L-glutamine modulates anxiety-like behavior, cortical spreading depression, and microglial immunoreactivity: analysis in developing rats suckled on normal size- and large size litters.

In mammals, L-glutamine (Gln) can alter the glutamate-Gln cycle and consequently brain excitability. Here, we investigated in developing rats the effect of treatment with different doses of Gln on anxiety-like behavior, cortical spreading depression (CSD), and microglial activation expressed as Iba1-immunoreactivity. Wistar rats were suckled in litters with 9 and 15 pups (groups L 9 and L 15; respectively, normal size- and large size litters). From postnatal days (P) 7-27, the animals received Gln per gavage (250, 500 or 750 mg/kg/day), or vehicle (water), or no treatment (naive). At P28 and P30, we tested the animals, respectively, in the elevated plus maze and open field. At P30-35, we measured CSD parameters (velocity of propagation, amplitude, and duration). Fixative-perfused brains were processed for microglial immunolabeling with anti-IBA-1 antibodies to analyze cortical microglia. Rats treated with Gln presented an anxiolytic behavior and accelerated CSD propagation when compared to the water- and naive control groups. Furthermore, CSD velocity was higher (p < 0.001) in the L 15 compared to the L 9 condition. Gln treatment increased Iba1 immunolabeling both in the parietal cortex and CA1 hippocampus, indicating microglial activation. The Gln effect was dose-dependent for anxiety-like behavior and CSD in both litter sizes, and for microglial activation in the L 15 groups. Besides confirming previous electrophysiological findings (CSD acceleration after Gln), our data demonstrate for the first time a behavioral and microglial activation that is associated with early Gln treatment in developing animals, and that is possibly operated via changes in brain excitability.

L-glutamine supplementation during the lactation period facilitates cortical spreading depression in well-nourished and early-malnourished rats.

AIMS: Glutamine (Gln) participates in the so-called "brain glutamine-glutamate cycle" and therefore it is likely to influence brain excitability. Here we investigated, in weaned well-nourished and early-malnourished rats, the effects of previous Gln oral supplementation, during the brain development period, on cortical spreading depression (CSD), an excitability-related brain phenomenon. MAIN METHODS: Male Wistar (W) suckling rat pups, well-nourished (litters with 6 pups) and malnourished (M) during lactation (by increasing the litters to 12 pups), received Gln (500 mg/kg/day) by gavage during postnatal days 7 to 27. At 30-40 days of life, they were submitted to a cortical spreading depression (CSD) recording session during 4 h, on 2 cortical parietal points of the right hemisphere. CSD velocity propagation was calculated from the time required for a CSD wave to cross the inter-electrode distance. KEY FINDINGS: In both nutritional condition, Gln rats presented higher (p<0.05) CSD propagation velocities (W-Gln, 4.22+/-0.23; M-Gln, 4.51+/-0.27 mm/min), as compared to water-treated controls (W-Wa, 3.77+/-0.21; M-Wa, 4.15+/-0.18 mm/min). This water control group did not differ from a naive control group that was not submitted to the gavage procedure. A fourth group, treated with a "placebo amino acid" (glycine), also displayed CSD velocities in the control range. SIGNIFICANCE: The results indicate that Gln supplementation during brain development facilitates cortical spreading depression propagation, as judged by the higher CSD velocities, and this effect is not abolished by malnutrition. Data support the idea of Gln-related changes in brain excitability, during neural development.

Fluoxetine inhibits cortical spreading depression in weaned and adult rats suckled under favorable and unfavorable lactation conditions.

Wistar rats (n = 58) were injected subcutaneously during the lactation period with fluoxetine (5, 10, 20 or 40 mg/kg/day) and cortical spreading depression (SD) was recorded immediately after weaning (25-30 days of life). An additional group (10 mg/kg; n = 8) was SD-recorded at 60-70 days. As compared to the saline-injected (n = 24) or "ingenuous" (n = 16) controls, fluoxetine dose-dependently reduced (P < 0.05) SD-velocities in the young rats by 4, 6, 16 and 15%, respectively, and in adult rats by 13%. In another experiment (26 adult rats), topical cortical application of fluoxetine (5 and 10 mg/ml solutions over the intact dura-mater for 10 min; n = 12 and 14, respectively) dose-dependently reduced SD-velocity (7.6% and 43.3% maximal reductions; P < 0.05). SD-propagation was blocked in 4 out of the 14 W-rats topically treated with the highest fluoxetine concentration (10 mg/ml). This topical fluoxetine effect was reverted after flushing the treated region with saline. In additional, 58 early-malnourished rats, fluoxetine applied during the suckling period (10 mg/kg/day, s.c.) and topically (10 mg/ml) also reduced (P < 0.05) SD-velocities by 18 and 22% for the systemic treatment (young and adult animals, respectively) and by 22.4% for the topical one. The present fluoxetine action supports the hypothesis of an antagonistic serotoninergic influence on SD, as previously suggested in experiments using other serotoninergic drugs. Data also suggest that early malnutrition does not greatly affect fluoxetine effects on SD.