BACKGROUND: Atrial natriuretic peptide, brain natriuretic peptide and C-type natriuretic peptide belong to a family of hormones that have natriuretic and vasodepressor activity and may play a pathophysiologic role in hypertension, heart failure and renal failure. Whereas immunoreactive human forms of these three natriuretic peptides are found in renal tubules, it is not clear whether they are derived from the systemic circulation or from local production. OBJECTIVE: To examine the gene expression of natriuretic peptides in cultured human glomerular cells. MATERIALS AND METHODS: We sought to determine the presence of messenger RNA encoding for these natriuretic peptides using polymerase chain reaction following reverse transcription. The polymerase chain reaction products were confirmed by direct sequencing. Atrial natriuretic peptide, brain natriuretic peptide and C-type natriuretic peptide in cell-culture supernatants were measured by radioimmunoassays (with detection limits of 2.1, 2.1 and 0.21 pmol/l, respectively). RESULTS: Atrial natriuretic peptide messenger RNA was not found in mesangial or glomerular epithelial cells (despite stimulation with tumor necrosis factor-alpha) except when the cells were cultured with a high concentration of fetal bovine serum (> 10%). Similarly, this peptide was not detected in supernatant unless the cells were cultured with fetal bovine serum at concentrations of > 10%. Brain natriuretic peptide messenger RNA was readily detected in cultured mesangial and glomerular epithelial cells with a lower concentration in the former. Brain natriuretic peptide was not found in the supernatant of resting mesangial cells but became detectable when incubated with tumor necrosis factor-alpha or fetal bovine serum. C-type natriuretic peptide messenger RNA was detected in mesangial and glomerular epithelial cells with a higher concentration in the latter. C-type natriuretic peptide was detected in the supernatant of resting glomerular epithelial cells and levels rose when incubated with increasing concentrations of tumor necrosis factor-alpha or fetal bovine serum. However, C-type natriuretic peptide was not detected in the supernatant of resting mesangial cells and remained undetectable following incubation with tumor necrosis factor-alpha or fetal bovine serum. CONCLUSION: Our results suggest differences in the synthesis of natriuretic peptides between glomerular mesangial and epithelial cells.
BACKGROUND: Atrial natriuretic peptide, brain natriuretic peptide and C-type natriuretic peptide belong to a family of hormones that have natriuretic and vasodepressor activity and may play a pathophysiologic role in hypertension, heart failure and renal failure. Whereas immunoreactive human forms of these three natriuretic peptides are found in renal tubules, it is not clear whether they are derived from the systemic circulation or from local production. OBJECTIVE: To examine the gene expression of natriuretic peptides in cultured human glomerular cells. MATERIALS AND METHODS: We sought to determine the presence of messenger RNA encoding for these natriuretic peptides using polymerase chain reaction following reverse transcription. The polymerase chain reaction products were confirmed by direct sequencing. Atrial natriuretic peptide, brain natriuretic peptide and C-type natriuretic peptide in cell-culture supernatants were measured by radioimmunoassays (with detection limits of 2.1, 2.1 and 0.21 pmol/l, respectively). RESULTS: Atrial natriuretic peptide messenger RNA was not found in mesangial or glomerular epithelial cells (despite stimulation with tumor necrosis factor-alpha) except when the cells were cultured with a high concentration of fetal bovine serum (> 10%). Similarly, this peptide was not detected in supernatant unless the cells were cultured with fetal bovine serum at concentrations of > 10%. Brain natriuretic peptide messenger RNA was readily detected in cultured mesangial and glomerular epithelial cells with a lower concentration in the former. Brain natriuretic peptide was not found in the supernatant of resting mesangial cells but became detectable when incubated with tumor necrosis factor-alpha or fetal bovine serum. C-type natriuretic peptide messenger RNA was detected in mesangial and glomerular epithelial cells with a higher concentration in the latter. C-type natriuretic peptide was detected in the supernatant of resting glomerular epithelial cells and levels rose when incubated with increasing concentrations of tumor necrosis factor-alpha or fetal bovine serum. However, C-type natriuretic peptide was not detected in the supernatant of resting mesangial cells and remained undetectable following incubation with tumor necrosis factor-alpha or fetal bovine serum. CONCLUSION: Our results suggest differences in the synthesis of natriuretic peptides between glomerular mesangial and epithelial cells.
Authors: S Jeson Sangaralingham; Denise M Heublein; Joseph P Grande; Alessandro Cataliotti; Andrew D Rule; Paul M McKie; Fernando L Martin; John C Burnett Journal: Am J Physiol Renal Physiol Date: 2011-08-24