Real-time imaging of renin release in vitro

János Peti-Peterdi, A. Fintha, Amanda L. Fuson, Albert Tousson, Robert H. Chow

Research output: Contribution to journalArticle

73 Citations (Scopus)

Abstract

Renin release from juxtaglomerular granular cells is considered the rate-limiting step in activation of the renin-angiotensin system that helps to maintain body salt and water balance. Available assays to measure renin release are complex, indirect, and work with significant internal errors. To directly visualize and study the dynamics of both the release and tissue activity of renin, we isolated and perfused afferent arterioles with attached glomeruli dissected from rabbit kidneys and used multiphoton fluorescence imaging. Acidotropic fluorophores, such as quinacrine and LysoTrackers, clearly and selectively labeled renin granules. Immunohistochemistry of mouse kidney with a specific renin antibody and quinacrine staining colocalized renin granules and quinacrine fluorescence. A low-salt diet for 1 wk caused an approximately fivefold increase in the number of both individual granules and renin-positive granular cells. Time-lapse imaging showed no signs of granule trafficking or any movement, only the dimming and disappearance of fluorescence from individual renin granules within 1 s in response to 100 μM isoproterenol. There appeared to be a quantal release of the granular contents; i.e., an all-or-none phenomenon. Using As4.1 cells, a granular cell line, we observed further classic signs of granule exocytosis, the emptying of granule content associated with a flash of quinacrine fluorescence. Using a fluorescence resonance energy transfer-based, 5-(2-aminoethylamino)naphthalene-1-sulfonic acid (EDANS)-conjugated renin substrate in the bath, an increase in EDANS fluorescence (renin activity) was observed around granular cells in response to isoproterenol. Fluorescence microscopy is an excellent tool for the further study of the mechanism, regulation, and dynamics of renin release.

Original languageEnglish
JournalAmerican Journal of Physiology - Renal Physiology
Volume287
Issue number2 56-2
DOIs
Publication statusPublished - Aug 2004

Fingerprint

Renin
Quinacrine
Fluorescence
Isoproterenol
Time-Lapse Imaging
In Vitro Techniques
Kidney
Sodium-Restricted Diet
Angiotensinogen
Fluorescence Resonance Energy Transfer
Sulfonic Acids
Body Water
Optical Imaging
Exocytosis
Arterioles
Renin-Angiotensin System
Baths
Fluorescence Microscopy
Salts
Immunohistochemistry

Keywords

  • Fluorescence microscopy
  • Juxtaglomerular apparatus
  • Multiphoton excitation
  • Quinacrine
  • Renin activity

ASJC Scopus subject areas

  • Physiology

Cite this

Real-time imaging of renin release in vitro. / Peti-Peterdi, János; Fintha, A.; Fuson, Amanda L.; Tousson, Albert; Chow, Robert H.

In: American Journal of Physiology - Renal Physiology, Vol. 287, No. 2 56-2, 08.2004.

Research output: Contribution to journalArticle

Peti-Peterdi, János ; Fintha, A. ; Fuson, Amanda L. ; Tousson, Albert ; Chow, Robert H. / Real-time imaging of renin release in vitro. In: American Journal of Physiology - Renal Physiology. 2004 ; Vol. 287, No. 2 56-2.
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