Separate K+ and Cl- transport pathways are activated for regulatory volume decrease in jejunal villus cells.
We assessed ion transport mechanisms operative during regulatory volume decrease (RVD) in jejunal villus enterocytes, isolated in suspension from guinea pig jejunum and examined with electronic cell sizing. Immediately after reduction of osmolarity (153 mosmol/kg medium) enterocytes swelled, but within 5 min they shrank by 50%. This RVD, which was complete by 20 min, was unaffected by Li+ substitution for Na+ or by Na(+)-free (N-methyl-D-glucose, NMDG+) medium. Passive loss of K+ is required for RVD because both the magnitude and direction of RVD changed when external [K+] varied. Increasing K+ permeability with gramicidin (0.5 microM) accelerated RVD in NMDG+ medium (10.0 +/- 0.8 vs. 6.2 +/- 0.4% min-1, P less than 0.01) suggesting that K+ loss is rate limiting for RVD. Inhibition of K(+)- and Ca2(+)-activated K+ conductance with Ba2+ (5 mM, P less than 0.005), quinine (100 microM, P less than 0.005), or apamin (1 microM, P less than 0.005) prevented RVD. Inhibition of Cl- conductance with 9-anthracenecarboxylic acid (100 microM, P less than 0.005) or dipyridamole (75 microM, P less than 0.005) also prevented RVD. In isotonic HCO3(-)-buffered medium, the addition of gramicidin to cells generated conditions in which anion permeability was rate limiting for cell swelling. This swelling was inhibited 97% by 100 microM 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS). In K(+)-free, HCO3(-)-buffered medium containing DIDS and gramicidin hypotonic swelling resulted in continued (secondary) swelling (rel vol 1.19 +/- 0.01 vs. 1.25 +/- 0.02, P less than 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)
MacLeod
R J
RJ
Department of Pediatrics, McGill University-Montreal Children's Hospital Research Institute, Quebec, Canada.
Hamilton
J R
JR
D000818:Animals; D000838:Anions; D001693:Biological Transport, Active; D002712:Chlorides / Q000378:metabolism*; D006096:Gramicidin / Q000494:pharmacology; D006168:Guinea Pigs; D006706:Homeostasis; D066298:In Vitro Techniques; D007583:Jejunum / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D007700:Kinetics; D008297:Male; D009130:Muscle, Smooth / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D009994:Osmolar Concentration; D010042:Ouabain / Q000494:pharmacology; D010539:Permeability; D011188:Potassium / Q000378:metabolism*; D011803:Quinine / Q000494:pharmacology
D000838:Anions; D002712:Chlorides; D006096:Gramicidin; D010042:Ouabain; D011803:Quinine; D011188:Potassium