The ion conductances of colonic crypts from dexamethasone-treated rats.
Whole-cell patch-clamp studies were performed in isolated colonic crypts of rats pretreated with dexamethasone (6 mg/kg subcutaneously on 3 days consecutively prior to the experiment). The cells were divided into three categories according to their position along the crypt axis: surface cells (s.c.); mid-crypt cells (m.c.) and crypt base cells (b.c.). The zero-current membrane voltage (Vm) was -56 +/- 2 mV in s.c (n = 34); -76 +/- 2 mV in m.c. (n = 47); and -87 +/- 1 mV in b.c. (n = 87). The whole-cell conductance (Gm) was similar (8-12 nS) in all three types of cells. A fractional K+ conductance accounting for 29-67% of Gm was present in all cell types. A Na+ conductance was demonstrable in s.c. by the hyperpolarizing effect on Vm of a low-Na+ (5 mmol/l) solution. In m.c. and b.c. the hyperpolarizing effect was much smaller, albeit significant. Amiloride had a concentration-dependent hyperpolarizing effect on Vm in m.c. and even more so in s.c.. It reduced Gm by approximately 12%. The dissociation constant (KD) was around 0.2 micromol/l. Triamterene had a comparable but not additive effect (KD = 30 micromol/l, n = 14). Forskolin (10 micromol/l, in order to enhance cytosolic adenosine 3', 5'-cyclic monophosphate or cAMP) depolarized Vm in all three types of cells. The strongest effect was seen in b.c.. Gm was enhanced significantly in b.c. by 83% (forskolin) to 121% [8-(4-chlorophenylthio)cAMP]. The depolarization of Vm and increase in Gm was caused to large extent by an increase in Cl-conductance as shown by the effect of a reduction in bath Cl-concentration from 145 to 32 mmol/l. This manoeuvre hyperpolarized Vm under control conditions significantly by 6-9 mV in all three types of cells, whilst it depolarized Vm in the presence of forskolin in m.c. and in b.c.. These data indicate that s.c. of dexamethasone-treated rats possess mostly a K+ conductance and an amiloride- and triamterene-inhibitable Na+ conductance. m.c. and b.c. possess little or no Na+ conductance; their Vm is largely determined by a K+ conductance. Forskolin (via cAMP) augments the Cl- conductance of m.c. and b.c. but has only a slight effect on s.c.
Ecke
D
D
Physiologisches Institut, Hermann Herder Strasse 7, D-79104 Freiburg, Germany.
Bleich
M
M
Schwartz
B
B
Fraser
G
G
Greger
R
R
D000584:Amiloride / Q000494:pharmacology; D000818:Animals; D002712:Chlorides / Q000378:metabolism; D003106:Colon / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D000242:Cyclic AMP / Q000494:pharmacology; D003907:Dexamethasone / Q000008:administration & dosage / Q000494:pharmacology*; D004553:Electric Conductivity; D004594:Electrophysiology; D066298:In Vitro Techniques; D007279:Injections, Subcutaneous; D007408:Intestinal Absorption; D018408:Patch-Clamp Techniques; D011188:Potassium / Q000502:physiology; D051381:Rats; D012964:Sodium / Q000493:pharmacokinetics*; D014223:Triamterene / Q000494:pharmacology
D002712:Chlorides; D000584:Amiloride; D003907:Dexamethasone; D012964:Sodium; D000242:Cyclic AMP; D011188:Potassium; D014223:Triamterene