Acidosis induces necrosis and apoptosis of cultured hippocampal neurons.
Acidosis, hypoxia, and hypoglycemia rapidly and transiently appear after reduction of cerebral blood flow. Acidosis also accompanies head trauma and subarachnoid hemorrhage. These insults result in necrotic and apoptotic loss of neurons. We previously demonstrated that transient acidification of intracellular pH from 7.3 to 6.5 induces delayed neuronal loss in cultured hippocampal slices (49). We now report that acidosis induced both necrotic and apoptotic loss of neurons. Necrosis and apoptosis were distinguished temporally and pharmacologically. Necrosis appeared rapidly and was dose dependent with the duration of the acidosis treatment. Apoptosis was delayed with maximal number of apoptotic cells seen with a 30-min acidosis treatment. Apoptotic neuronal loss was accompanied by DNA fragmentation and was blocked by inhibitors of protein and RNA synthesis, ectopic expression of the anti-apoptotic gene bcl-2, or an inhibitor of caspases, proteases known to be activated during apoptosis. Necrotic neuronal loss was unaffected by these treatments. Hypothermia, a treatment known to attenuate neuronal loss following a variety of insults, blocked both acidosis-induced necrosis and apoptosis. These results indicate that acidosis is neurotoxic in vitro and suggest that acidosis contributes to both necrotic and apoptotic neuronal loss in vivo.
Ding
D
D
Department of Pharmacology and Physiology, State University of New York-Downstate Medical Center, Brooklyn, New York, 11203, USA.
Moskowitz
S I
SI
Li
R
R
Lee
S B
SB
Esteban
M
M
Tomaselli
K
K
Chan
J
J
Bergold
P J
PJ
D000138:Acidosis / Q000473:pathology*; D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D002545:Brain Ischemia / Q000503:physiopathology / Q000628:therapy; D020169:Caspases / Q000378:metabolism; D003080:Cold Temperature; D003470:Culture Media / Q000494:pharmacology; D018076:DNA, Complementary / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D015870:Gene Expression / Q000502:physiology; D006624:Hippocampus / Q000166:cytology*; D006863:Hydrogen-Ion Concentration; D007036:Hypothermia, Induced; D020287:In Situ Nick-End Labeling; D009336:Necrosis; D009410:Nerve Degeneration / Q000503:physiopathology; D009474:Neurons / Q000201:enzymology / Q000473:pathology*; D009924:Organ Culture Techniques; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley
D003470:Culture Media; D018076:DNA, Complementary; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D020169:Caspases