Expression of inter-α inhibitor related proteins in kidneys and urine of hyperoxaluric rats

Manabu T. Moriyama, Patricia A. Glenton, Saeed R. Khan

Research output: Contribution to journalArticle

Abstract

Purpose: To investigate the involvement of the inter-α inhibitor family of proteins in calcium oxalate stone formation we determined immunohistochemical distribution in the kidneys and excretion in the urine of these proteins in normal and hyperoxaluric rats. Various members of the family have been shown to inhibit the formation and retention of calcium oxalate crystals in the kidneys. Materials and Methods: Hyperoxaluria was induced in male Sprague-Dawley rats by administering 0.75% ethylene glycol. The inter-α inhibitor family consists of inter-α inhibitor, pre-α inhibitor, the so-called heavy chains H1, H2 and H3, and the light chain bikunin. Antibodies against these molecules were used to localize various proteins in rat kidneys by immunohisto-chemical techniques. Urine was analyzed by sodium dodecyl sulfate-polyacrylamide gel electro-phoresis and Western blot analysis to determine the expression of various members of the inter-α inhibitor family. Results: In normal kidneys staining for inter-α inhibitor and other members of the family was mostly limited to the proximal tubules and generally to their luminal contents. Eight weeks after the induction of hyperoxaluria various sections of renal tubules stained positive for inter-α inhibitor, bikunin and H3. Positive staining was observed in the tubular lumina as well as in the cytoplasm of epithelial cells. Crystal associated material was heavily stained. Western blot analysis recognized 7 protein bands in the urine. The urinary expression of H1, H3 and pre-α-inhibitor was significantly increased. Conclusions: Apparently hyperoxaluria and renal calcium oxalate crystal deposition result in the increased expression of crystallization inhibitors, such as inter-α-inhibitor related proteins, in the kidneys and urine. Results indicate that kidneys respond to nephrolithic challenges by producing proteins that inhibit crystal formation and retention.

Original languageEnglish (US)
Pages (from-to)1687-1692
Number of pages6
JournalJournal of Urology
Volume165
Issue number5 I
StatePublished - 2001
Externally publishedYes

Fingerprint

Urine
Kidney
Hyperoxaluria
Calcium Oxalate
Proteins
Western Blotting
Staining and Labeling
Ethylene Glycol
Crystallization
Sodium Dodecyl Sulfate
Sprague Dawley Rats
Cytoplasm
Epithelial Cells
Light
Antibodies

Keywords

  • Calcium oxalate
  • Hyperoxaluria
  • Kidney
  • Rats
  • Sprague-Dawley
  • Urinary calculi

ASJC Scopus subject areas

  • Urology

Cite this

Moriyama, M. T., Glenton, P. A., & Khan, S. R. (2001). Expression of inter-α inhibitor related proteins in kidneys and urine of hyperoxaluric rats. Journal of Urology, 165(5 I), 1687-1692.

Expression of inter-α inhibitor related proteins in kidneys and urine of hyperoxaluric rats. / Moriyama, Manabu T.; Glenton, Patricia A.; Khan, Saeed R.

In: Journal of Urology, Vol. 165, No. 5 I, 2001, p. 1687-1692.

Research output: Contribution to journalArticle

Moriyama, MT, Glenton, PA & Khan, SR 2001, 'Expression of inter-α inhibitor related proteins in kidneys and urine of hyperoxaluric rats', Journal of Urology, vol. 165, no. 5 I, pp. 1687-1692.
Moriyama, Manabu T. ; Glenton, Patricia A. ; Khan, Saeed R. / Expression of inter-α inhibitor related proteins in kidneys and urine of hyperoxaluric rats. In: Journal of Urology. 2001 ; Vol. 165, No. 5 I. pp. 1687-1692.
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N2 - Purpose: To investigate the involvement of the inter-α inhibitor family of proteins in calcium oxalate stone formation we determined immunohistochemical distribution in the kidneys and excretion in the urine of these proteins in normal and hyperoxaluric rats. Various members of the family have been shown to inhibit the formation and retention of calcium oxalate crystals in the kidneys. Materials and Methods: Hyperoxaluria was induced in male Sprague-Dawley rats by administering 0.75% ethylene glycol. The inter-α inhibitor family consists of inter-α inhibitor, pre-α inhibitor, the so-called heavy chains H1, H2 and H3, and the light chain bikunin. Antibodies against these molecules were used to localize various proteins in rat kidneys by immunohisto-chemical techniques. Urine was analyzed by sodium dodecyl sulfate-polyacrylamide gel electro-phoresis and Western blot analysis to determine the expression of various members of the inter-α inhibitor family. Results: In normal kidneys staining for inter-α inhibitor and other members of the family was mostly limited to the proximal tubules and generally to their luminal contents. Eight weeks after the induction of hyperoxaluria various sections of renal tubules stained positive for inter-α inhibitor, bikunin and H3. Positive staining was observed in the tubular lumina as well as in the cytoplasm of epithelial cells. Crystal associated material was heavily stained. Western blot analysis recognized 7 protein bands in the urine. The urinary expression of H1, H3 and pre-α-inhibitor was significantly increased. Conclusions: Apparently hyperoxaluria and renal calcium oxalate crystal deposition result in the increased expression of crystallization inhibitors, such as inter-α-inhibitor related proteins, in the kidneys and urine. Results indicate that kidneys respond to nephrolithic challenges by producing proteins that inhibit crystal formation and retention.

AB - Purpose: To investigate the involvement of the inter-α inhibitor family of proteins in calcium oxalate stone formation we determined immunohistochemical distribution in the kidneys and excretion in the urine of these proteins in normal and hyperoxaluric rats. Various members of the family have been shown to inhibit the formation and retention of calcium oxalate crystals in the kidneys. Materials and Methods: Hyperoxaluria was induced in male Sprague-Dawley rats by administering 0.75% ethylene glycol. The inter-α inhibitor family consists of inter-α inhibitor, pre-α inhibitor, the so-called heavy chains H1, H2 and H3, and the light chain bikunin. Antibodies against these molecules were used to localize various proteins in rat kidneys by immunohisto-chemical techniques. Urine was analyzed by sodium dodecyl sulfate-polyacrylamide gel electro-phoresis and Western blot analysis to determine the expression of various members of the inter-α inhibitor family. Results: In normal kidneys staining for inter-α inhibitor and other members of the family was mostly limited to the proximal tubules and generally to their luminal contents. Eight weeks after the induction of hyperoxaluria various sections of renal tubules stained positive for inter-α inhibitor, bikunin and H3. Positive staining was observed in the tubular lumina as well as in the cytoplasm of epithelial cells. Crystal associated material was heavily stained. Western blot analysis recognized 7 protein bands in the urine. The urinary expression of H1, H3 and pre-α-inhibitor was significantly increased. Conclusions: Apparently hyperoxaluria and renal calcium oxalate crystal deposition result in the increased expression of crystallization inhibitors, such as inter-α-inhibitor related proteins, in the kidneys and urine. Results indicate that kidneys respond to nephrolithic challenges by producing proteins that inhibit crystal formation and retention.

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