Measurement of the cytosolic sodium ion concentration in rat brain synaptosomes by a fluorescence method

Abstract

A method for the measurement of the cytosolic Na+ concentration in intact synaptosomes is described. This method makes use of a pH sensitive dye (BCECF) that can be loaded into the cytosol and a relatively specific ionophore (monensin) that can exchange Na+ for H+ across the synaptosomal membrane. By setting conditions such that there is no electrochemical potential difference for H+ across the membrane (no membrane potential and pHi = pHo), addition of ionophore would induce a H+ flux only if there is a concentration difference for Na+. Thus, when there is no fluorescence change (no cytosolic pH change) extracellular [Na+] equals intrasynaptosomal [Na+]. The intrasynaptosomal [Na+] concentration was determined to be 7 +/- 3 mM (n = 5; mean +/- S.E.). The results obtained with this fluorescence method are compared with estimates obtained by atomic absorption spectrometry. Limitations and applications of the method are discussed.