An ultrastructural study of the kidney of normal, copper poisoned and thiomolybdate-treated sheep

Histological, ultrastructural and kidney function techniques were used to assess changes in the kidney of sheep given either copper (Cu) or Cu and the Cu complexing agent thiomolybdate (TM), or TM alone. Kidney function was normal in sheep given Cu and TM together or TM alone. In these animals the cells lining cortical tubules accumulated Cu within numerous, large, electron-dense lysosomes. Sheep given Cu alone developed haemolysis, impaired kidney function and a variety of morphological defects including an increase in number of large lysosomes in cells of the cortical tubules. There was a breakdown of the glomerular endothelial lining and fusion of foot processes. Cells of the cortical tubules showed degeneration and necrosis and an increase in microbodies and rough endoplasmic reticulum. Cortical and medullary blood vessels were dilated, with evidence of breakdown of the endothelial lining. Copper appeared to injure kidney tissue at three sites, tubular epithelium, glomerular basal lamina and capillary blood vessels. Changes reported here are similar to the renal lesions in cadmium toxicity

Distribution of copper in the kidney of sheep after thiomolybdate administration

Chronic copper poisoning (CCP) is a well recognized harzard of sheep production (1). There is evidence that the incidence of CCP is increasing as more intensive methods of sheep production are adopted (2). In CCP, haemolysis is the most characteristic symptom and hence the disease has been categorized into 3 phases; pre-haemolytic, haemolytic and post-haemolytic (3, 4). Animals may develop several haemolytic crises despite the removal of the source of excess copper (Cu) at the time of the first haemolysis (5). Kidney damage occurs during haemolysis and persists into the post-haemolytic phase. Gooneratne et al. (6) have shown that thiomolybdate (TM) injections prevented the occurrence of the haemolytic crisis when given to Cu loaded sheep before haemolysis and were of value in treating sheep in which the haemolysis of CCP had already occurred. However, the kidneys of TM treated animals accumulated large amounts of Cu. The consequences of this were not known. The intracellular distribution of Cu in the kidney of either animals or humans has not been reported previously. The present investigation was carried out to determine the pattern of distribution of Cu within the subcellular fractions of the kidney in normal, TM treated and Cu loaded-TM treated sheep. Acid phosphatase (AP) activity was measured to identify the movement of lysosomes within the fractions as our previous studies on the kidney (7) indicated that there was an increase in lysosomes within the tubule cells of the kidney of experimental chronic copper poisoned sheep

Distribution of Cu, Zn, and Fe in the soluble fraction of the kidney in normal, copper-poisoned, and thiomolybdate-treated sheep

Twenty-seven sheep were used in two experiments to study the distribution of copper (Cu), zinc (Zn), and iron (Fe) in the kidney cytosol of control, Cu-loaded, and thiomolybdate (TM)-treated sheep. A comparison was made of the patterns of distribution on Sephadex G-75 of Cu, Zn, and Fe from the cytosol of fresh and frozen kidneys and after thawing of frozen cytosol. In both Cu-dosed and TM-treated sheep, the absolute level of Cu increased in the cytosol, but the percent of Cu decreased. The percent recovery of Cu from the frozen kidney was comparable to that from fresh kidney, but the extraction of Cu from the supernatant of frozen cytosol was approximately 10% less. This was due to a loss of Cu in a precipitate that formed when the frozen cytosol was thawed. Most of the Cu in the cytosol from the kidney of Cu-loaded sheep was in a metallothionein (MT)-like protein fraction and was trichloroacetic acid (TCA) soluble. In contrast, that from the cytosol of TM-treated sheep was mostly in a high molecular weight fraction that was TCA insoluble. The chromatograms obtained from cytosol derived from frozen kidneys, or cytosol that had itself been frozen, contained a similar distribution of Cu, Zn, and Fe, but the peak heights were lower in the latter samples

Subcellular distribution of copper in the kidneys of normal, copper-poisoned, and thiomolybdate-treated sheep

Twenty-seven sheep given either copper (Cu) and/or tetrathiomolybdate (TM) were used to study the subcellular distribution of Cu within the kidney and to monitor the location of lysosomes within the subcellular fractions using acid phosphatase (AP) as a marker enzyme. Copper dosing alone increased the Cu content in the liver and the kidneys. The administration of intravenous TM prevented the development of chronic copper poisoning (CCP) in sheep, reduced the rate of accumulation of Cu in the liver of Cu-dosed animals, but increased the Cu content of kidneys in both the control and Cu-dosed sheep. The total amount of Cu that accumulated in the kidneys of sheep given TM appears to depend on several factors: a) liver Cu concentration, b) Cu intake, and c) dosage of TM. Thus, the highest Cu concentration was found in the kidneys of sheep that continued to receive Cu orally at the same time as they were given TM. The intracellular distribution of Cu and AP in the kidneys showed that in the control sheep given neither Cu or TM, the highest proportion of Cu was in the cytosol fraction, and the highest specific activity of AP was in the light mitochondrial (lysosomal) fraction. Dosing with Cu markedly increased the Cu concentration and greatly elevated the total activity of AP in the heavier fractions, i.e., the nuclear (N) and heavy mitochondrial (MH). Thus, the increase in Cu observed in the N and MH fractions was not caused by an accumulation of Cu by nuclei and mitochondria, but was due to an accumulation of Cu by lysosomes that sedimented with the heavier fractions. The intracellular distribution of Cu in the kidneys of TM-treated sheep was similar to that seen in Cu-loaded sheep. Although Cu accumulated readily in the kidneys of animals receiving TM, kidney function tests showed neither glomerular nor tubular functional impairment