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Sodium-coupled glycine uptake by Ehrlich ascites tumor cells results in an increase in cell volume and plasma membrane channel activities.
Authors:R L Hudson and S G Schultz
Affiliation:Department of Physiology and Cell Biology, University of Texas Medical School, Houston 77225.
Abstract:The addition of 10 mM glycine to a physiological saline bathing Ehrlich ascites tumor cells is followed by a slow increase in cell volume that plateaus between 15 and 30 min at a level approximately equal to 17% greater than the control volume; this increase is not observed when glycine is added to cells suspended in a Na+-free saline. The results of studies using the patch-clamp technique in the cell-attached mode indicate that, 0.5-3 min after the addition of glycine to the bathing solution, there is a marked increase in the activity of single channels, which is almost all instances were previously present and operant in the plasma membrane. Successfully excised patches of membrane that contained a channel stimulated by glycine fell into two categories. Some became inactive within 15 sec in spite of the fact that the G omega seal remained intact. Others persisted for the lifetime of the seal. All of the persistent channels had an 11-fold selectivity for Cl- over K+ and a conductance of 23 pS when bathed by symmetrical 150 mM KCl solutions. Although the ionic specificities of the other channels have not been identified, there is reason to suspect that they might be K+ channels whose activities are dependent on factors lost when the patch is excised. Swelling induced by exposing these cells to a 50% hypotonic perfusate stimulated the activities of Cl- channels whose properties closely resemble those stimulated by the addition of glycine to the perfusate, strongly suggesting that the glycine-induced stimulation of Cl- channel activity is part of a volume-regulatory response to cell swelling. If the increase in channel activity induced by the addition of glycine to the perfusate is indeed a response to cell swelling, then this perfusate is indeed a response to cell swelling, then this volume-regulatory response must be extremely sensitive inasmuch as it appears to be "triggered" by an average increase in cell volume that does not exceed 5%.
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