Subcellular distribution of copper in the liver of fetal deer in the last month of gestation

Abstract

Fetal deer, in the last month of gestation, accumulate high concentrations of copper in the liver. Livers from fetal deer in late gestation were homogenised and fractionated by continuous sucrose density gradient centrifugation. The distribution of copper closely followed that of DNA; approximately two thirds of the metal was localised to the nuclear fractions with the remaining third in the cytosolic fractions. The fractionation procedure was repeated with digitonin, a lysosomal perturbant: lysosomes were disrupted and the marker enzyme, N-acetyl-(3-glucosaminidase, shifted from the particulate fractions to the cytosolic fractions; the distribution of copper was unaffected. Differential centrifugation of homogenate confirmed that approximately two thirds of the copper was associated with the nuclear fraction. Further confirmation of a nuclear localisation of copper was provided by X-ray microanalysis of purified nuclei.