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Effects of high calcium, low pH on immature versus adult rat brain mitochondria


, : Effects of high calcium, low pH on immature versus adult rat brain mitochondria. Society for Neuroscience Abstracts 27(1): 301

Although age-related differences in brain energy metabolism and mitochondrial enzyme activities are established, less is known about the effects of age on mitochondrial responses to factors that contribute to brain injury after ischemia or trauma. This study tested the hypothesis that synaptosomal plus non-synaptosomal brain mitochondria (mito) isolated from normal immature rats (16-18 do) would respond differently to in vitro exposure to high calcium (Ca) and low pH than brain mito from normal adult rats. Respiration-dependent mito Ca uptake was measured by fluorescent detection of the decrease in Ca added to a potassium-based medium with and without ATP (3 mM), at a pH of 7.0 and 6.5. Mito suspensions were analyzed for release of cytochrome c (CytC) in response to Ca or a pro-apoptotic, BH3 cell death domain-containing peptide (60 uM). Acidosis caused a reduction in Ca uptake in both immature and adult mito, which was more pronounced in immature rats (80% less vs 40% less in adult mito). In the presence of ATP, adult mito exhibited greater Ca uptake at either pH. In contrast, without ATP, immature mito exhibited a greater Ca uptake than adult mito at both pH 7.0 (487 vs 217 nmol Ca/mg) and pH 6.5 (441 vs 166 nmol Ca/mg). Immunoblots showed Ca-dependent CytC release from both immature and adult mito. BH3 peptide-induced release was seen only in immature mito, consistent with the presence of endogenous Bax in immature mito. These differences in the responses of immature and adult brain mito to Ca, acidic pH and a BH3 peptide may influence the relative contributions of these factors to the pathobiology of secondary ischemic and traumatic brain injury.

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