Mitochondrial structure and function in the untreated Jackson toxic milk (tx-j) mouse, a model for Wilson disease.

Structural changes in hepatocellular mitochondria are characteristic of Wilson disease (WD). Features include variability in size and shape, increased density of matrix, discreet inclusions, and cystic dilatation of the cristae. We examined the functional basis for these mitochondrial changes in the toxic milk (tx-j) mouse model for WD. Its normal syngeic strain, C3H, served as control. Hepatic histology was near-normal in tx-j mice at 3-4-months-old and showed mild inflammation and steatosis at 6-months-old. Transmission electron microscopy showed typical mitochondrial abnormalities, specifically cystic dilatation of tips of cristae, in 3, 4, and 6-month-old tx-j mice and none in normal 3-month-old C3H mice. Citrate synthase (CS) activity was initially lower in tx-j mice than age-matched controls but increased over the first 6 months such that it was significantly greater at 5 and 6-months-old (p<0.003). No evidence for hepatic mtDNA depletion was found by long-PCR analysis. NB-PAGE showed preservation of all complexes in the oxidative-phosphorylation chain except complex IV which declined markedly from 5-months-old onwards. Hepatic complex IV activity was significantly decreased in 5-month-old tx-j mice (p<0.04). Expression of mitochondrial transfer factor A (TFAM) mRNA declined progressively in 6-8-month-old tx-j mice; immunodetectable protein levels declined in parallel. Expression of mtSSB mRNA was uniformly low in tx-j mice from 1-8-months-old. Levels of two mitochondrial antioxidant proteins capable of binding copper, thioredoxin-2 and peroxiredoxin-3, rose over the first 6 months of life. Mitochondrial changes occur early in WD and reflect complex, probably oxidative, injury.