Osmotic Sensitivity of Taurine Release from Hippocampal Neuronal and Glial Cells

Abstract

Taurine transport is important for volume regulation ofcultured neurons and astroglial cells. Both cell types utilize similar mechanisms for taurine accumulation and efflux. However, taurine lost from cerebellar Purkinje cells in vivo is accumulated by adjacent astrocytes during hypoosmotic hyponatremia. To examine mechanisms for transfer of taurine between cell types, we measured relative sensitivities of taurine loss from cultured neurons and astrocytes. Primary cultures of hippocampal neurons and astrocytes were grown from embryonic and neonatal rat brain, respectively.Neurons were used after 10–14 days in culture. Astrocytes were used after 14 days in culture and were grown in the same culture medium used for neurons for 3 days prior toexperimentation. Cells were incubated at 37∘C for 30 min in isoosmotic (290 mOsm) phosphate-buffered saline (PBS). The PBS was then changed to fresh isoosmotic or to hypoosmotic PBS (270 mOsm or 250 mOsm), made by reducing the NaCl concentration. Cell volume and taurine content were determined immediately before hypoosmotic exposure or 3, 15, or 30 min later. In isoosmotic PBS, astrocytes contained 162 ± 18 nmol taurine/mg protein (mean ± SEM), equivalent to an intracellular concentration of 30.2 ± 2.1 mM. No taurine loss was detectable after 3 or 15 min in either hypoosmotic PBS, but after 30 min in 270 or 250 mOsm PBS, astrocyte taurine was reduced by 8.0% or 22.2%, respectively. Neurons initially contained 114 ± 13 nmol taurine/mg protein, equivalent to an intracellular taurine concentration of 22.2 ± 2.5 mM. After 3 min of exposure to 270 or 250 mOsm PBS, the cells had lost 17 ±% or 25 ±% of their taurine content,respectively. Cell volumes of each cell type were similarly affected by hypoosmotic exposure. We conclude that taurine loss from cultured hippocampal neurons is more sensitive to osmotic swelling than taurine loss from cultured hippocampal astrocytes. This characteristic, if present in cells ofthe hippocampus in vivo, could lead to net transfer oftaurine from neurons to glial cells during pathological conditions which cause cell swelling.