Abstract
Metallothionein (Mt) is a low molecular weight metalloprotein and its synthesis is induced by various divalent metals (Cd, Zn, Hg and Cu). The Intracellular distribution of Mt in hepatic and renal cells from control and CdCl 2 injected rats was investigated by immunohistochemical methods. Antiserum to purified rat-liver Mt was prepared in rabbits after partial polymerization of the protein. The unlabelled peroxidase-antiperoxidase method using the specific rabbit anti-rat liver Mt provided a sensitive technique to localize Mt in tissue sections. In control rats, Mt or thionein (metal-free protein) was mainly localized in the cytoplasm of hepatocytes, renal collecting duct epithelium and distal convoluted tubular epithelium. In rats, injected intraperitoneally (i.p.) with CdCl 2 (0.6 mg/kg) for 2 weeks, Mt was present mainly in the nuclei which were largely negative in control rats. Repeated injection with CdCl 2 for 4–8 weeks resulted in the appearance of Mt in both the nucleus and cytoplasm. Intraluminal staining was also noted in proximal convoluted tubules along with marked vacuolation in the cytoplasm at 6 and 8 weeks. High intensity staining was observed in proximal convoluted tubules and collecting duct epithelium of the kidneys in CdCl 2 injected rats. The bile duct epithelium in liver samples, renal glomerular mesangial cells, glomerular visceral epithelial cells and vascular smooth muscle cells showed weak to moderately intense staining. No staining was seen in vascular endothelial cells, fibroblasts or leukocytes. The staining for Mt in this technique was abolished when the antibody was absorbed with rat-liver Mt in vitro or by substitution of the antiserum with normal rabbit serum, demonstrating the specificity of the staining reaction for Mt. The results showed the presence of small amounts of Mt predominantly in the cytoplasm of control rat hepatocytes and renal tubular epithelium and its appearance in both the nucleus and cytoplasm after its synthesis induced by CdCl 2 injections.
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