Tumor necrosis factor-alpha mRNA expression in lipopolysaccharide-stimulated rat kidney. Chronological analysis of localization

Am J Pathol. 1994 Jun;144(6):1159-66.

Abstract

To study the time course of lipopolysaccharide-induced production of tumor necrosis factor-alpha (TNF-alpha) in the kidney, we utilized a highly sensitive non-radioisotopic in situ hybridization with 1-nm gold-conjugated anti-digoxigenin for localization of TNF-alpha mRNA expression after lipopolysaccharide administration. TNF-alpha mRNA expression localized by in situ hybridization showed a peak increment in proximal tubular epithelial cells and glomeruli at 2 hours and returned to almost normal levels at 6 hours. The intensity of the signal was much stronger in proximal tubules than in glomeruli. These findings were confirmed by the demonstration of similar kinetics in the increase of TNF-alpha message, measured by using amplification of the third and fourth exons of TNF-alpha gene by reverse transcription-polymerase chain reaction of microdissected proximal tubular segments and isolated glomeruli. Reverse transcription-polymerase chain reaction of cultured rat mesangial and glomerular epithelial cells demonstrated that mesangial cells, not glomerular epithelial cells, were responsible for the observed glomerular signals.

MeSH terms

  • Animals
  • Base Sequence
  • Cells, Cultured
  • DNA / analysis
  • DNA / genetics
  • Epithelium / chemistry
  • Female
  • Glomerular Mesangium / chemistry
  • In Situ Hybridization
  • Kidney / chemistry*
  • Kidney / cytology
  • Lipopolysaccharides / pharmacology*
  • Molecular Sequence Data
  • Nephrons / chemistry
  • Polymerase Chain Reaction
  • RNA, Messenger / analysis*
  • RNA, Messenger / genetics
  • Rats
  • Rats, Sprague-Dawley
  • Tumor Necrosis Factor-alpha / analysis
  • Tumor Necrosis Factor-alpha / genetics*

Substances

  • Lipopolysaccharides
  • RNA, Messenger
  • Tumor Necrosis Factor-alpha
  • DNA