The mechanism of spermidine release from Xenopus oocytes was examined by measuring release of radioactive [3H]spermidine under different ionic conditions, and under voltage-clamp. In normal solution (2 mm K+), the efflux rate is less than 1% per hour, and is stimulated ∼2-fold by inclusion of Ca2+ (1 mM) in the incubation medium. Spermidine efflux is stimulated ∼10-fold in high [K+] (KD98) solution. In KD98 solution, efflux is strongly inhibited by divalent cations (Ki for Ba2+ block of spermidine efflux is ∼0.1 mM), but not by tetraethyl-ammonium ions or verapamil. Spermidine efflux rates were not different between control oocytes and those expressing HRK1 inward rectifier K+ (Kir) channels. When the membrane potential was clamped, either by changing external [K+] in oocytes expressing HRK1, or by 2-microelectrode voltage-clamp, spermidine efflux was shown to be strongly dependent on voltage, as expected for a simple electrodiffusive process, where spermidine3+ is the effluxing species. This result argues against spermidine diffusing out as an uncharged species, or in exchange for similarly charged counterions. These results are the first conclusive demonstration of a simple electrodiffusive pathway for spermidine efflux from cells.

Original languageEnglish
Pages (from-to)3392-3397
Number of pages6
JournalJournal of Biological Chemistry
Issue number7
StatePublished - Feb 16 1996


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