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  • Title: Hypertonic shrinking but not hypotonic swelling increases sodium concentration in rat brain synaptosomes.
    Author: Waseem TV, Kolos VA, Lapatsina LP, Fedorovich SV.
    Journal: Brain Res Bull; 2007 Jun 15; 73(1-3):135-42. PubMed ID: 17499647.
    Abstract:
    Neurotransmitter release is dependent on both calcium and sodium influx. Hypotonic swelling and hypertonic shrinking of neurons evokes calcium-independent exocytosis of neurotransmitters into the synaptic cleft. To date, there are not too much data available on relationship between extracellular osmolarity and sodium concentration in presynaptic endings. In the present study we investigated the effects of hypotonic swelling and hypertonic shrinking on sodium levels, as measured using fluorescent dyes SBFI-AM and Sodium Green in rat brain synaptosomes. Reduction of incubation medium osmolarity from 310 to 230 mOsm did not raise the intrasynaptosomal sodium concentration. An increase of osmolarity from 310 to 810 mOsm is accompanied by a dose-dependent elevation of sodium concentration from 8.1+/-0.5 to 46.5+/-2.8mM, respectively. This effect was insensitive to several channel inhibitors such as: tetrodotoxin, an inhibitor of voltage-gated sodium channels, bumetanide, an inhibitor of Na(+)/K(+)/2Cl(-) cotransport, gadolinium, an inhibitor of nonselective mechanosensitive channels, ruthenium red, an inhibitor of transient receptor potential channel and amiloride, an inhibitor of epithelial sodium channel/degenerin. Additionally, using the fluorescent dye BCECF-AM, we have shown that hypertonic shrinking caused a dose-dependent acidification of intrasynaptosomal cytosol, which suggests that the Na(+)/H(+) exchanger is not involved in the effect of increased osmolarity on cytosolic sodium levels. The increase in intrasynaptosomal sodium concentrations following increases in osmolarity is probably due to sodium influx through another sodium channels.
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