Subject(s)
Kidney Glomerulus/physiology , Regeneration/genetics , Animals , Glial Cell Line-Derived Neurotrophic Factor/physiology , Humans , Interleukin-6/physiology , Kidney Glomerulus/cytology , Kidney Glomerulus/embryology , Leukemia Inhibitory Factor , Proteoglycans/physiology , Proto-Oncogene Proteins/physiology , Proto-Oncogene Proteins c-sis/physiology , Transcription Factors/physiology , Transforming Growth Factor beta/physiology , Vascular Endothelial Growth Factor A/physiology , Wnt Proteins/physiology , Wnt4 ProteinABSTRACT
BACKGROUND: Peptidylarginine deiminases (PADs) are a group of posttranslational modification enzymes that citrullinate (deiminate) protein arginine residues, yielding citrulline residues. Citrullination of arginine residues abolishes their positive charge, markedly altering their structure. We undertook this study to investigate the actions of PADs in the kidney. METHODS: In male rats, we ligated the unilateral ureter, then analyzed the obstructed and contralateral kidneys 1 week later. Controls were rats simultaneously given sham operations. In another experiment, we ligated unilateral ureters of eight rats, four of which received a ureter-bladder anastomosis 1 week later. These rats were subjected to histologic examinations 5 weeks after unilateral ureteral obstruction (UUO). RESULTS: Reverse transcription-polymerase chain reaction (RT-PCR) revealed that, of PADs (type I, II, III, and IV), only PAD type II was expressed in kidneys. Western blot study showed that PAD type II expression and citrullinated protein content increased greatly in kidneys that underwent unilateral ureteral ligation compared to that in contralateral or sham-operated kidneys. Immunohistochemical analyses revealed that PAD type II was preferentially expressed by parietal epithelial cells and that only in Bowman's capsule were proteins citrullinated. Additionally, these PAD type II and citrullinated proteins in obstructed nephropathy were significantly attenuated by the release of the obstruction. Proteome analysis revealed that one of citrullinated proteins in the kidney should be actin. CONCLUSION: This result indicates that PAD type II and citrullinated proteins are suitable markers of Bowman's capsule. Not only are these markers preferentially expressed in Bowman's capsules but their expression is also increased in damaged kidneys by UUO, features that promise the further clarification of kidney diseases.
Subject(s)
Citrulline/metabolism , Kidney Glomerulus/metabolism , Kidney Glomerulus/physiopathology , Ureteral Obstruction/metabolism , Ureteral Obstruction/physiopathology , Actin Cytoskeleton/metabolism , Animals , Biomarkers , Blood Urea Nitrogen , Creatinine/blood , Epithelial Cells/enzymology , Hydrolases/genetics , Male , Peptide Mapping , Protein-Arginine Deiminase Type 2 , RNA, Messenger/analysis , Rats , Rats, Sprague-DawleyABSTRACT
We examined (CTG)n lengths in various tissues from a 70-year-old man with myotonic dystrophy type 1 (DM 1) who had a small 60-70 (CTG), expansion in his leukocytes. He died of renal cell carcinoma 5 years after a total laryngectomy for laryngeal carcinoma. Southern blot and polymerase chain reaction analyses were done on tissues obtained at autopsy. In the various normal tissues, (CTG). lengths were almost all the same size, whereas the renal cell carcinoma and metastatic tissues had longer lengths. When compared with the lengths in leukocytes about 5 years previously, (CTG)n lengths in the normal tissues were the same size. These findings suggest that both somatic instability and age-dependent (CTG). expansion in DM 1 patients with a small expansion may be less dominant than in patients with large expansions.
