The Journal of Membrane Biology

, Volume 94, Issue 2, pp 99–115 | Cite as

Electrogenic properties of the sodium-alanine cotransporter in pancreatic acinar cells: I. Tight-seal whole-cell recordings

  • P. Jauch
  • O. H. Petersen
  • P. Läuger
Articles
Received:

Summary

Electrical currents associated with sodium-coupled alanine transport in mouse pancreatic acinar cells were studied using the method of whole-cell recording with patch pipettes. Single cells or small clusters of (electrically coupled) cells were isolated by collagenase treatment. The composition of the intracellular solution could be controlled by internal perfusion of the patch pipette. In this way both inward and outward currents could be measured under “zero-trans” conditions, i.e., with finite concentrations of sodium andl-alanine on one side and zero concentrations on the other. Inward andoutward currents for equal but opposite concentration gradients were found to be of similar magnitude, meaning that the cotransporter is functionally nearly symmetric. The dependence of current on the concentrations of sodium andl-alanine exhibited a Michaelis-Menten behavior. From the sodium-concentration dependence of current as well as from the reversal potential of the current in the presence of an alanine-concentration, gradient, a sodium/alanine stoichiometric ratio of 1:1 can be inferred. The finding that N-methylated amino acids may substitute, forl-alanine, as well as the observed pH dependence of currents indicate that the pancreatic alanine transport system is similar to (or identical with) the “A-system” which is widespread in animal cells. The transport system is tightly coupled with respect to Na+; alanine-coupled inward flow of Na+ is at least 30 times higher than uncoupled Na+ flow mediated by the cotransporter. The current-voltage characteristic of the cotransporter could be (approximately) determined from the difference of transmembrane current in the presence and in the absence ofl-alanine. The sodium-concentration dependence of the current-voltage characteristic indicates that a Na+ ion approaching the binding site from the extracellular medium has to cross part of the transmembrane electric field.

Key words

cotransport electrogenic transport sodium-coupled amino-acid transport pancreatic acinar cells whole-cell recording 
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Copyright information

© Springer-Verlag New York Inc 1986

Authors and Affiliations

  • P. Jauch
    • 1
  • O. H. Petersen
    • 2
  • P. Läuger
    • 1
  1. 1.Department of BiologyUniversity of KonstanzKonstanzFederal Republic of Germany
  2. 2.The Physiological LaboratoryUniversity of LiverpoolLiverpoolUK

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