Growth factors prevent mitochondrial dysfunction, loss of calcium homeostasis, and cell injury, but not ATP depletion in hippocampal neurons deprived of glucose.
The mechanism of growth factor protection against metabolic/excitotoxic insults was examined. The time course of changes in ATP levels, mitochondrial transmembrane potential, intracellular free calcium levels ([Ca2+]i), and cell survival resulting from glucose deprivation were assessed in cultured hippocampal neurons. ATP levels were significantly reduced within 1 h of the onset of glucose deprivation and reached less than 20% of control levels by 12 h. Mitochondrial transmembrane potential (assessed by rhodamine 123 accumulation in mitochondria) declined progressively between 4 and 20 h following the onset of glucose deprivation. The [Ca2+]i was reduced during the first 1 h of glucose deprivation, gradually rose through 12 h, and then rose rapidly and was elevated five- to sevenfold after 16 h. The [Ca2+]i did not increase, and mitochondrial dysfunction and cell damage were prevented, in hypoglycemic neurons incubated in Ca(2+)-deficient medium. Elevation of [Ca2+]i by exposure of neurons to glutamate caused loss of rhodamine 123 fluorescence and structural damage to mitochondria. Mitochondrial function could be restored and cell survival maintained by addition of glucose prior to the late elevation of [Ca2+]i. Nerve growth factor (NGF), basic fibroblast growth factor (bFGF), and insulin-like growth factor II (IGF-II) prevented loss of both [Ca2+]i homeostasis and mitochondrial transmembrane potential, and protected hippocampal neurons against hypoglycemic injury, but did not prevent the hypoglycemia-induced reduction in ATP levels. NaCN and 2,4-dinitrophenol (DNP) caused a large elevation of [Ca2+]i, mitochondrial dysfunction, and cell death. NGF, bFGF, and IGF-II each significantly reduced the adverse effects of NaCN and DNP on [Ca2+]i, mitochondrial function, and cell survival. Loss of [Ca2+]i homeostasis may be a critical event leading to mitochondrial damage and cell death resulting from energy failure. Preventing loss of [Ca2+]i homeostasis may be a general mechanism for the neuroprotective action of growth factors.
Sanders-Brown Research Center on Aging, Lexington 40536-0230.
M P
MP
Mattson
Y
Y
Zhang
S
S
Bose
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000502:physiology*; D002470:Cell Survival / Q000187:drug effects; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005947:Glucose / Q000172:deficiency*; D006133:Growth Substances / Q000494:pharmacology*; D006624:Hippocampus / Q000378:metabolism* / Q000473:pathology; D006706:Homeostasis / Q000187:drug effects*; D007003:Hypoglycemia / Q000473:pathology / Q000503:physiopathology*; D007335:Insulin-Like Growth Factor II / Q000494:pharmacology; D008564:Membrane Potentials; D008928:Mitochondria / Q000187:drug effects / Q000502:physiology*; D009414:Nerve Growth Factors / Q000494:pharmacology; D009474:Neurons / Q000187:drug effects*; D051381:Rats; D012966:Sodium Cyanide / Q000009:adverse effects
D006133:Growth Substances; D009414:Nerve Growth Factors; D016222:Fibroblast Growth Factor 2; D007335:Insulin-Like Growth Factor II; D000255:Adenosine Triphosphate; D005947:Glucose; D012966:Sodium Cyanide; D002118:Calcium