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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

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Abstract

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.

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Acknowledgements

The authors thank the following Brazilian agencies for financial support: Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq No. 445101/2014-8), Instituto Brasileiro de Neurociências (IBN-Net/Finep No. 4191), and Capes (Edital 043/2013 Ciências Do Mar II). RCA Guedes is a Research Fellow from CNPq (No. 303636/2014-9).

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  1. Department of Nutrition, Universidade Federal de Pernambuco, Recife, PE, 50670901, Brazil

    Denise Sandrelly Cavalcanti de Lima, Elian da Silva Francisco, Cássia Borges Lima & Rubem Carlos Araújo Guedes

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  1. Denise Sandrelly Cavalcanti de Lima
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  2. Elian da Silva Francisco
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  3. Cássia Borges Lima
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de Lima, D.S.C., Francisco, E.d., Lima, C.B. et al. 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. Amino Acids 49, 337–346 (2017). https://doi.org/10.1007/s00726-016-2365-2

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