cAMP mediates ammonia-induced programmed cell death in the microglial cell line BV-2.
Although ammonia is a well-known neuropathogenic factor, the cellular mechanisms of ammonia toxicity are less characterized. Up to now, the main focus of ammonia toxicity has been on astrocytes and neurons. Despite the significance of microglia in neurodegenerative diseases, little is known about their responsiveness to ammonia. In the present study, we found that ammonia triggered mitosis at concentrations between 30 microm and 3.0 mm but apoptosis at concentrations >or= 1.0 mm in the murine microglial cell line BV-2. Most apoptotic cells showed an accumulation of condensed chromatin at the nuclear envelope, blebbing of the plasma membrane, formation of apoptotic bodies and an increase in caspase 3/7 activity. Blockade of caspase 3/7 activity by Ac-DEVD-CHO suppressed ammonia-induced apoptosis. Surprisingly, some BV-2 cells exposed to ammonia displayed clear signs of mitotic catastrophe, a type of cell death occurring during mitosis. In a further series of experiments, we found that cyclic adenosine 3',5'-monophosphate (cAMP) mediated the apoptogenic effects of ammonia, because (i) ammonia dose-dependently elevated the intracellular cAMP level, (ii) blockade of the adenylyl cyclase by SQ-22536 suppressed ammonia-induced apoptosis, (iii) inhibition of phosphodiesterases (PDEs) by the nonselective PDE inhibitor IBMX, or by the PDE4-selective inhibitor rolipram, increased the relative number of apoptotic cells, and (iv) the cAMP analogues 8-bromoadenosine cAMP and Sp-cAMP mimicked the effect of ammonia and triggered apoptosis. Taken together, our results indicate that distinct concentrations of ammonia trigger opposite signalling pathways in microglial cells.
Svoboda
Nina
N
Department of Cell Biology, University of Salzburg, Hellbrunnerstr. 34, 5020 Salzburg, Austria.
Zierler
Susanna
S
Kerschbaum
Hubert H
HH
D000641:Ammonia / Q000633:toxicity*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D002460:Cell Line; D048430:Cell Shape / Q000187:drug effects; D000242:Cyclic AMP / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D051379:Mice; D017628:Microglia / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D008938:Mitosis / Q000187:drug effects; D015398:Signal Transduction / Q000502:physiology
D000641:Ammonia; D000242:Cyclic AMP; D053148:Caspase 3; D053179:Caspase 7