Astrocytes protect neurons from ammonia toxicity.
Ammonia is a neurotoxin that is implicated in the CNS dysfunction associated with hepatic encephalopathy, urea cycle disorders, Reye's syndrome and other neurological conditions. While in vivo studies suggest that astrocytes are the principal target of ammonia toxicity, recent in vitro investigations suggest that neurons may also be directly affected by ammonia. To further examine the issue of neural cell sensitivity to ammonia, pure rat cortical neuronal cultures, as well as co-cultures of neurons and astrocytes, were exposed to 5 mM NH4Cl for 48 h. Cultures were examined for morphological changes by light microscopy, measures of cell death, free radical production and changes in the mitochondrial inner membrane potential. Ammonia caused extensive degenerative changes in pure cultured neurons, while such neuronal changes were minor in the co-cultures. Similarly, processes of pure cultured neurons displayed a significant loss of the mitochondrial inner membrane potential, as compared to neurons in co-cultures. Cell death (LDH release) in ammonia-treated neuronal cultures was twice as great as untreated controls, while in co-cultures ammonia did not significantly increase cell death. Free radical production at 3 min was increased (69%, P<0.05) in pure neuronal cultures but not in co-cultures. The neuroprotective effects observed in co-cultures may have been mediated by the astrocyte's ability to scavenge free radicals, by their detoxification of ammonia and/or by their neurotrophic actions. The neuroprotective action of astrocytes may explain the failure to detect significant pathological changes in neurons in ammonia toxicity in vivo.
Rao
K V Rama
KV
Veterans Affairs Medical Center, University of Miami, School of Medicine, Miami, FL 33101, USA.
Panickar
K S
KS
Jayakumar
A R
AR
Norenberg
M D
MD
D000641:Ammonia / Q000633:toxicity*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001253:Astrocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002478:Cells, Cultured; D018920:Coculture Techniques; D005609:Free Radicals / Q000378:metabolism; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008564:Membrane Potentials / Q000502:physiology; D008928:Mitochondria / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D051381:Rats
D005609:Free Radicals; D000641:Ammonia; D007770:L-Lactate Dehydrogenase; C505407:CASP3 protein, human; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases