Photon correlation spectroscopy of brain mitochondrial populations: Application to traumatic brain injury

Mitochondrial dysfunction and pathology that contribute to a host of neurodegenerative diseases are deduced from changes in ultrastructure, routinely examined by a host of optical techniques. We adapted the technique of photon correlation spectroscopy (PCS) to evaluate calcium-induced structural alterations in isolated viable cortical and hippocampal mitochondria. In detecting calcium-induced reductions in light intensity, PCS was more sensitive than absorbance across varying calcium concentrations. Mitochondrial populations encompass a broad distribution of sizes, confirmed by ultrastructural profiles, both which remain unaffected by calcium exposure. Cortical and hippocampal populations show fractional calcium-induced reductions in light scatter compared to subsequent maximal alamethicin-induced reductions. Although reductions in light scatter (refractive index) have been interpreted as mitochondrial swelling, PCS quantification of the mean mitochondrial radius demonstrates that mitochondrial size is unaffected by calcium exposure, but not alamethicin. Likewise, the population distribution histograms remain stable with calcium exposure, but shift to larger radii after alamethicin exposure. Furthermore, hippocampal mitochondrial populations from a neurodegenerative model of traumatic brain injury, lateral fluid percussion, demonstrate greater calcium-induced reductions in scatter intensity, which are associated with an initial population of large mitochondria becoming smaller. The disparate responses to calcium and subsequent alamethicin of mitochondria at 3 and 24 h after injury attest to an acute disruption of membrane permeability in mitochondria from injured brain. PCS provides quantitative indices of refractive index and size in isolated mitochondrial populations, aiding the evaluation of mitochondria in degenerative diseases.