The action of 5-hydroxytryptamine on normal and cystic fibrosis mouse colon: effects on secretion and intracellular calcium.

The ability of mouse colon to generate a secretory response to stimulation by 5-hydroxytryptamine (5-HT) was investigated in intact colonic sheets mounted in Ussing chambers. A preparation of intact isolated crypts was used to determine whether 5-HT action was associated with an elevation of cytosolic calcium levels, measured using the calcium-sensitive fluorescent dye, fura-2. 5-HT increased the short-circuit current, an effect that was inhibited by 55% in the absence of chloride and by 83% in the presence of serosal frusemide, consistent with the stimulation of electrogenic chloride secretion. This was confirmed by the observation that colonic tissue from transgenic cystic fibrosis mice (n = 4) failed to respond to 5-HT, although wild-type tissues generated an increased short-circuit current of 52.4+/-1.1 microAcm(-2) (n = 9). The electrical response to 5-HT was calcium-dependent. 5-HT action was unaffected by tetrodotoxin and was not mimicked by the 5-HT3 agonist 1-phenylbiguanide, indicating that neural mechanisms are not involved. The cyclooxygenase inhibitor indomethacin, however, reduced the 5-HT-induced rise in short-circuit current by 73%, suggesting that prostaglandin production contributes to the response. Stimulation of crypts with acetylcholine elicited an increase in cytosolic calcium levels, but no such response was detected on application of 5-HT (10(-6) to 10(-4) M), suggesting that 5-HT does not directly modulate intracellular calcium in colonic crypt cells. It is concluded that mouse colon responds to 5-HT challenge with a stimulation of electrogenic chloride secretion and that this effect is mediated by indirect mechanisms that might involve immune elements within the colonic wall.