Stress protein synthesis induced by cadmium-cysteine in rat kidney.
Biomarkers are important tools which enable toxicologists to reliably predict and detect exposures to xenobiotics and resultant cell injury, ultimately improving risk assessments. Since the de novo synthesis of stress proteins can be detected early after exposure to some agents, analysis of toxicant-induced changes in gene expression, i.e. alterations in patterns of protein synthesis, may be useful to develop as biomarkers of exposure and toxicity. We are utilizing various xenobiotics as tools to study stress protein synthesis in target organs in order to evaluate the target tissue-specificity of this response. Previous data from this laboratory have demonstrated that induction of stress proteins in rat liver, but not kidney, after acute exposure to CdCl2 precedes hepatoxicity. Since kidney is a target tissue after chronic Cd exposure, it was of interest to examine stress protein synthesis in this tissue. However, dose-limiting hepatotoxicity precluded this evaluation. Cd complexed with molecules such as cysteine (cys) or metallothionein has been used in acute dosing regimens as a tool in order to study the nephrotoxicity of Cd. Therefore, this study was undertaken in order to evaluate Cd-induced stress protein synthesis in an important tissue known to be injured after chronic exposure, i.e. kidney. Specific objectives included comparing stress protein synthesis in rat kidney and liver after acute exposure to Cd-cys and CdCl2, determining the Cd threshold concentration for renal stress protein synthesis and assessing the relationship between stress protein synthesis and nephropathy. Male rats were exposed to equivalent doses of Cd as CdCl2 or Cd-cysteine (molar ratio Cd:cys = 1:15). Kidney Cd concentrations increased 5-fold after i.v. injection of Cd-cys compared to CdCl2, mimicking Cd distribution following chronic exposure. After exposure to Cd, tissue slices were incubated with 35S-methionine. Slices were subsequently homogenized and centrifuged, and the 16,000 g supernatants were subjected to SDS-polyacrylamide gel electrophoresis. Proteins which had incorporated 35S-methionine were detected by autoradiography. De novo synthesis of 70, 90 and 110 kDa proteins was enhanced in liver, but not in kidney, 4 h after injection of 2 mg Cd/kg as CdCl2. In contrast, dose-related increases in synthesis of these proteins were observed in kidney 4 h after injection of 1 and 2mg Cd/kg as Cd-cys, but not at lower dosages. In addition, synthesis of a 68 kDa kidney protein was inhibited at 2 mg Cd/kg as Cd-cys. The threshold for Cd-induced stress protein synthesis was shown to be between 4 and 8 micrograms Cd/g tissue.(ABSTRACT TRUNCATED AT 400 WORDS)
Goering
P L
PL
Health Sciences Branch, Center for Devices and Radiological Health, Food and Drug Administration, Rockville, Maryland 20857.
Kish
C L
CL
Fisher
B R
BR
D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002104:Cadmium / Q000008:administration & dosage / Q000378:metabolism / Q000633:toxicity*; D019256:Cadmium Chloride; D002712:Chlorides / Q000008:administration & dosage / Q000378:metabolism / Q000633:toxicity; D003545:Cysteine; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D006360:Heat-Shock Proteins / Q000032:analysis / Q000096:biosynthesis*; D007668:Kidney / Q000187:drug effects* / Q000378:metabolism*; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley
D015415:Biomarkers; D002712:Chlorides; D006360:Heat-Shock Proteins; D002104:Cadmium; D019256:Cadmium Chloride; D003545:Cysteine