Chronic hyperammonemia impairs the glutamate–nitric oxide–cyclic GMP pathway in cerebellar neurons in culture and in the rat in vivo
Article first published online: 25 DEC 2001
European Journal of Neuroscience
Volume 10, Issue 10, pages 3201–3209, October 1998
How to Cite
Hermenegildo, C., Montoliu, C., Llansola, M., Muñoz, M.-D., Gaztelu, J.-M., Miñana, M.-D. and Felipo, V. (1998), Chronic hyperammonemia impairs the glutamate–nitric oxide–cyclic GMP pathway in cerebellar neurons in culture and in the rat in vivo. European Journal of Neuroscience, 10: 3201–3209. doi: 10.1046/j.1460-9568.1998.00329.x
- Issue published online: 25 DEC 2001
- Article first published online: 25 DEC 2001
- Received 9 December 1997, revised 16 May 1998, accepted 27 May 1998
- nitric oxide synthase;
- NMDA receptor
The aim of this work was to assess whether ammonia concentrations similar to the increase found in the brain of hyperammonemic rats (100 μm), impair N-methyl-d-aspartate (NMDA) receptor-mediated signal transduction. We first measured glutamate neurotoxicity, which in these neurons is mediated by activation of NMDA receptors, as an initial parameter reflecting activation of NMDA receptor-mediated pathways. Long-term treatment of cultured neurons with ammonia prevents glutamate-induced neuronal death. The EC50 was 20 μm, and at 100 μm the protection was complete. The induction of the protective effect was not immediate, but took several hours.
Treatment with 100 μm ammonia did not prevent a glutamate- or NMDA-induced rise of intracellular calcium. Ammonia impaired the glutamate–nitric oxide–cGMP (3′,5′-cyclic guanosine monophosphate) pathway in a dose- and time-dependent manner. Glutamate-induced formation of cGMP was reduced by 42%, while activation of nitric oxide synthase was not affected. Ammonia reduced by 31% cGMP formation induced by S-nitroso-N-acetyl-penicillamine (SNAP), a NO-generating agent, confirming that the interference occurs at the level of guanylate cyclase activation by nitric oxide.
To assess whether chronic moderate hyperammonemia in vivo also impairs the glutamate–nitric oxide–cGMP pathway, we determined by in vivo brain microdialysis in freely moving rats the formation of cGMP induced by NMDA. In hyperammonemic rats, the formation of cGMP induced by NMDA and SNAP was reduced by ca. 60 and 41%, respectively, indicating that chronic hyperammonemia in the animal in vivo also impairs the glutamate–nitric oxide–cGMP pathway. Impairment of this pathway can contribute to the neurological alterations found in hyperammonemia and hepatic encephalopathy.