The effect of secretagogues on ion conductances of in vitro perfused, isolated rabbit colonic crypts.

Abstract

Several secretagogues were used in this study, including those which enhance intracellular cyclic adenosine monophosphate (cAMP) production, as well as others which elevate intracellular Ca2+ activity and are known to increase Cl- secretion in the intact colon and in colonic carcinoma cell lines. They were examined with respect to their effects on electrophysiological properties in isolated rabbit distal colonic crypts. Crypts were dissected manually and perfused in vitro. Transepithelial voltage (Vte), transepithelial resistance (Rte), membrane voltage across the basolateral membrane (Vbl), and fractional basolateral membrane resistance (FRbl), were estimated. Basolateral prostaglandin E2 (PGE2, > or = 0.1 mumol/l), vasoactive intestinal peptide (VIP, 1 nmol/l) and adenosine (0.1 mmol/l) induced an initial depolarisation and a secondary partial repolarisation of Vbl. In the case of adenosine, the initial depolarization of Vbl was by 31 +/- 2 mV (n = 47). Rte fell significantly from 16.4 +/- 3.6 to 14.2 +/- 3.7 omega.cm2 (n = 6), and FRbl increased significantly from 0.11 +/- 0.02 to 0.51 +/- 0.10 (n = 6). In the second phase the repolarisation of Vbl amounted 11 +/- 2 mV (n = 47) and a steady-state (Vbl) of -51 +/- 2 mV (n = 47) was reached. Rte fell further and significantly to a steady-state value of 12.4 +/- 3.8 omega.cm2 (n = 6) and FRbl fell significantly to 0.42 +/- 0.13 (n = 6). In 30% of the experiments, a transient hyperpolarisation of Vbl by 8 +/- 2 mV (n = 14) was seen during wash out of adenosine.(ABSTRACT TRUNCATED AT 250 WORDS)