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  • Title: The ion conductances of colonic crypts from dexamethasone-treated rats.
    Author: Ecke D, Bleich M, Schwartz B, Fraser G, Greger R.
    Journal: Pflugers Arch; 1996 Jan; 431(3):419-26. PubMed ID: 8584437.
    Abstract:
    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.
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