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ATP stimulation of volume regulation in cultured human astrocytoma cells
Author(s) -
Woods Megan,
Olson James
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.lb514
Subject(s) - osmole , osmotic concentration , purinergic receptor , stimulation , chemistry , receptor , cell , volume (thermodynamics) , cell culture , andrology , endocrinology , biology , biochemistry , medicine , physics , genetics , quantum mechanics
Understanding cell volume control of brain cells will lead to better treatments for patients suffering from brain edema following trauma and other conditions. We examined the role P2Y receptors play in cell volume regulation using cultured 1321N1 human astrocytoma cells. Methods Cells were removed from the growth surface and suspended in isoosmotic phosphate buffered saline (Iso‐PBS) containing (in mM): NaCl (147), KH 2 PO 4 (0.5), Na 2 HPO 4 (3.2), CaCl 2 (1), MgCl 2 (0.5), dextrose (5), and KCl (2.7) (pH=7.3, osmolality=290 mOsm) at 37 °C for 30 min. Cell volumes were determined using the Coulter method at various times after cells were resuspended in Iso‐PBS or similar solutions with osmolalities altered by changing concentrations of NaCl. Results We observed a linear relationship of initial volume versus (osm) −1 for cells in osmolalities ranging from 150 mOsm to 600 mOsm with an intercept at 45.3±1.3% of the volume measured in Iso‐PBS. Maximal cell swelling in 150 mOsm PBS occurred 3–5 min after suspension. After 10 min and 30 min in this hypoosmotic condition, cell volume had recovered by 18.3±5.1% and 48.4±2.2%, respectively. When 100 μM ATP was added to the 150 mOsm PBS solution, the rate of volume recovery was significantly faster. Conclusion 132N1 cells demonstrate regulatory volume decrease in hypoosmotic conditions which may be stimulated by purinergic receptors. Supported by NIH R25GM086257.