Two-photon in vivo microscopy of sulfonefluorescein secretion in normal and cystic rat kidneys

George A. Tanner, Ruben M. Sandoval, Kenneth Dunn

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Sulfonefluorescein (SF) is a fluorescent organic anion secreted by kidney proximal tubules. The purposes of this study were 1) to quantify accumulation of SF in normal and cystic rat kidneys in vivo and 2) to test whether SF accumulation could be used as a marker for cysts derived from proximal tubules. Male Munich-Wistar rats, normal Han:SPRD rats, and heterozygous Han:SPRD rats with autosomal-dominant polycystic kidney disease were anesthetized with Inactin and solutions containing SF were administered by constant intravenous infusion. In Munich-Wistar rats, SF fluorescence in the urinary space of Bowman's capsule averaged 0.15 ± 0.04 (n = 17) times that of glomerular capillary plasma, consistent with extensive plasma protein binding of SF. In normal Han:SPRD rats, steady-state cell cytoplasm SF fluorescence in proximal tubule and distal tubule cells averaged, respectively, 2.7 ± 1.4 (n = 99 tubules) and 0.2 ± 0.2 (n = 17) times that of peritubular capillary plasma. No punctate SF fluorescence was seen in proximal tubule cell cytoplasm. Probenecid reduced proximal tubule cell SF fluorescence to 0.64 ± 0.40 (n = 64) times that of plasma. Ureteral obstruction decreased the proximal tubule cell-to-lumen SF fluorescence gradient, suggesting that tubule fluid flow normally sweeps away secreted SF. In cystic kidneys, cysts derived from proximal tubules could be identified by their uptake of SF, but cell uptake was patchy. We conclude that in vivo two-photon microscopy is a powerful tool for quantifying glomerular and tubular handling of SF, and SF can be used to identify proximal tubule-derived cysts.

Original languageEnglish
JournalAmerican Journal of Physiology - Renal Physiology
Volume286
Issue number1 55-1
StatePublished - Jan 2004

Fingerprint

Cystic Kidney Diseases
Photons
Fluorescence
Cysts
sulfonefluorescein
Intravital Microscopy
Wistar Rats
Cytoplasm
Bowman Capsule
Autosomal Dominant Polycystic Kidney
Probenecid
Ureteral Obstruction
Proximal Kidney Tubule
Intravenous Infusions
Protein Binding

Keywords

  • Glomerulus
  • Organic anion secretion
  • Polycystic kidney disease
  • Two-photon microscopy

ASJC Scopus subject areas

  • Physiology

Cite this

Two-photon in vivo microscopy of sulfonefluorescein secretion in normal and cystic rat kidneys. / Tanner, George A.; Sandoval, Ruben M.; Dunn, Kenneth.

In: American Journal of Physiology - Renal Physiology, Vol. 286, No. 1 55-1, 01.2004.

Research output: Contribution to journalArticle

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N2 - Sulfonefluorescein (SF) is a fluorescent organic anion secreted by kidney proximal tubules. The purposes of this study were 1) to quantify accumulation of SF in normal and cystic rat kidneys in vivo and 2) to test whether SF accumulation could be used as a marker for cysts derived from proximal tubules. Male Munich-Wistar rats, normal Han:SPRD rats, and heterozygous Han:SPRD rats with autosomal-dominant polycystic kidney disease were anesthetized with Inactin and solutions containing SF were administered by constant intravenous infusion. In Munich-Wistar rats, SF fluorescence in the urinary space of Bowman's capsule averaged 0.15 ± 0.04 (n = 17) times that of glomerular capillary plasma, consistent with extensive plasma protein binding of SF. In normal Han:SPRD rats, steady-state cell cytoplasm SF fluorescence in proximal tubule and distal tubule cells averaged, respectively, 2.7 ± 1.4 (n = 99 tubules) and 0.2 ± 0.2 (n = 17) times that of peritubular capillary plasma. No punctate SF fluorescence was seen in proximal tubule cell cytoplasm. Probenecid reduced proximal tubule cell SF fluorescence to 0.64 ± 0.40 (n = 64) times that of plasma. Ureteral obstruction decreased the proximal tubule cell-to-lumen SF fluorescence gradient, suggesting that tubule fluid flow normally sweeps away secreted SF. In cystic kidneys, cysts derived from proximal tubules could be identified by their uptake of SF, but cell uptake was patchy. We conclude that in vivo two-photon microscopy is a powerful tool for quantifying glomerular and tubular handling of SF, and SF can be used to identify proximal tubule-derived cysts.

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