NBD-TMA: A novel fluorescent substrate of the peritubular organic cation transporter of renal proximal tubules

Dallas Bednarczyk, Eugene A. Mash, Bhasker Reddy Aavula, Stephen H. Wright

Research output: Contribution to journalArticle

33 Scopus citations

Abstract

Traditionally, the measurement of transport activity has employed radiolabeled compounds. The resulting experimental procedures do not measure transport in real time and are limited in temporal and spatial resolution. The use of epifluorescence microscopy provides the ability to measure transport activity in real time with high temporal and spatial resolution. Using epifluorescence microscopy we characterized the transport of the fluorescent organic cation, [2-(4-nitro-2,1,3-benzoxadiazol-7- yl)aminoethyl]trimethylammonium (NBD-TMA+, MW 266). NBD-TMA+ has structural characteristics common to other secreted organic cations and is fluorescent (λ(ex)=458 nm; λ(em)=530 nm). The excitation and emission spectra are insensitive to changes in [Cl-] and minimally sensitive to pH in the physiologically relevant range (pH 5.0-7.4). A microscope equipped with a photon-detection system was used to measure accumulation of NBD-TMA+ by isolated rabbit renal proximal tubules. Accumulation of NBD-TMA+ by proximal tubules was time dependent and saturable (Michaelis-Menten constant K(m) 12 μM). Proximal tubule accumulation of NBD-TMA+ was inhibited by the organic cations tetraethylammonium (TEA+) (apparent inhibitory constant K(app) (TEA) 134 μM), cimetidine, and N1-methylnicotinamide (NMN). Our experimental results provide strong evidence that NBD-TMA+ is transported by one or more of the basolateral organic cation transporters involved in the renal secretion of this chemical class of compound. This fluorescent substrate provides a sensitive means of investigating organic cation transport.

Original languageEnglish (US)
Pages (from-to)184-192
Number of pages9
JournalPflugers Archiv European Journal of Physiology
Volume440
Issue number1
DOIs
StatePublished - May 26 2000

Keywords

  • Fluorescence
  • Kidney
  • Tetraethylammonium
  • Transport

ASJC Scopus subject areas

  • Physiology
  • Clinical Biochemistry
  • Physiology (medical)

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