Glutaraldehyde fixation of sodium transport in dog red blood cells
The large increase in passive Na flux that occurs when dog red blood cells are caused to shrink is amiloride sensitive and inhibited when Cl is replaced by nitrate or thiocyanate. Activation and deactivation of this transport pathway by manipulation of cell volume is reversible. Brief treatment of the cells with 0.01-0.03% glutaraldehyde can cause the shrinkage-activated transporter to become irreversibly activated or inactivated, depending on the volume of the cells at the time of glutaraldehyde exposure. Thus, if glutaraldehyde is applied when the cells are shrunken, the amiloride-sensitive Na transporter is activated and remains so regardless of subsequent alterations in cell volume. If the fixative is applied to swollen cells, no amount of subsequent shrinkage will turn on the Na pathway. In its fixed state, the activated transporter is fully amiloride sensitive, but it is no longer inhibited when Cl is replaced by thiocyanate. The action of glutaraldehyde thus allows one to dissect the response to cell shrinkage into two phases. Activation of the pathway is affected by anions and is not prevented by amiloride. Once activated and fixed, the anion requirement disappears. Amiloride inhibits movement of Na through the activated transporter. These experiments demonstrate how a chemical cross-linking agent may be used to study the functional properties of a regulable transport pathway.
- Research Organization:
- Univ. of North Carolina School of Medicine, Chapel Hill
- OSTI ID:
- 5832955
- Journal Information:
- J. Gen. Physiol.; (United States), Vol. 84:5
- Country of Publication:
- United States
- Language:
- English
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ALDEHYDES
BIOCHEMICAL REACTION KINETICS
DIURETICS
SODIUM
BIOLOGICAL PATHWAYS
CHLORIDES
DOGS
ERYTHROCYTES
HYPERTONIC SOLUTIONS
INHIBITION
NITRATES
POTASSIUM
RADIOISOTOPES
THIOCYANATES
ALKALI METALS
ANIMALS
ANTITHYROID DRUGS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARBONIC ACID DERIVATIVES
CHLORINE COMPOUNDS
DISPERSIONS
DRUGS
ELEMENTS
HALIDES
HALOGEN COMPOUNDS
HORMONE ANTAGONISTS
ISOTOPES
KINETICS
MAMMALS
MATERIALS
METALS
MIXTURES
NITROGEN COMPOUNDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
OXYGEN COMPOUNDS
REACTION KINETICS
SOLUTIONS
VERTEBRATES
551001* - Physiological Systems- Tracer Techniques