A comparative study of functional 5‐HT4 receptors in human colon, rat oesophagus and rat ileum
Authors: McLean Peter G. , Coupar Ian M. , Molenaar Peter 
Published: July 2012

The pharmacological properties of 5‐hydroxytryptamine (5‐HT), the 5‐HT4 receptor agonists, DAU 6236 and SC 53116 and the 5‐HT4 receptor antagonist, GR 113808, were studied in the rat oesophagus, rat ileum and human colon.
5‐HT relaxed the longitudinal muscle of the rat oesophagus and rat ileum and the circular muscle of the human colon. Absolute values of relaxation were measured and showed the order of the maximum responses, rat oesophagus >>human colon > rat ileum with EC50 values of 189 ± 15 nM, 157±4nM, 306 ± 72 nM, respectively. 5‐HT also inhibited the spontaneous contractions of the human colon with an EC50 value of 119± 1 nM. The effect of 5‐HT on the human colon was not affected by methysergide (10 μM) or ondansetron (1 μAM).
The use of the uptake and metabolism inhibitors, cocaine (30μM) and pargyline (100μM), did not increase the potency of 5‐HT in the rat oesophagus or human colon. In the rat oesophagus, cocaine (30 μM) produced a reduction in carbachol‐induced tone of 22.2 ± 0.6% and reduced the 5‐HT maximum effect by 52.0 ± 0.4%.
The compounds, DAU 6236 and SC 53116, showed a different pattern of potencies and efficacies in the rat oesophagus, rat ileum and human colon compared to 5‐HT. DAU 6236 relaxed the human colonic circular muscle with an EC50 value of 129 ± 16 nM but its efficacy was less than that of 5‐HT. DAU 6236 (1 μM) also antagonized the 5‐HT‐induced relaxation of the human colon with a dose‐ratio of 9.9. In the rat oesophagus and rat ileum, DAU 6236 was inactive in the majority of tissues. In the minority of oesophagus tissues that did respond the EC50 value was 1.2 ±0.7μM. DAU 6236 also antagonized the effect of 5‐HT in the rat oesophagus in a non‐surmountable fashion. SC 53116 relaxed the rat oesophagus with an EC50 value of 91 ± 4 nM, with an efficacy less than that observed to 5‐HT; however, at 200 nM it did not antagonize the 5‐HT‐induced relaxation of the rat oesophagus. SC 53116 showed no agonist activity in the rat ileum and human colon, but at 1 μM it did antagonize the effect of 5‐HT in the human colon with a dose‐ratio of 11.3±0.3.
GR 113808 competitively antagonized the 5‐HT4 receptor‐mediated relaxation of the rat oesophagus with a pA2 value of 8.59 (8.18‐9.00) against 5‐HT and 9.05 (8.79‐9.31) against SC 53116. GR 113808 (0.01 μM) also antagonized the 5‐HT‐induced relaxation of human colonic circular muscle with an apparent pA2 value of 9.02 ±0.12. However at 1 μM the apparent pA2 value was significantly lower than that measured at 0.01 and 0.1 μM. GR 113808 (0.01 μM) antagonized the 5‐HT4 receptor‐mediated relaxation of the rat ileum with an apparent pA2 value of 9.30 ±0.21.
In conclusion, these studies have shown that the human colon, rat oesophagus and rat ileum contain functional 5‐HT4 receptors. However, the 5‐HT4 receptor agonists displayed differences in these tissues making it necessary to be cautious when extrapolating from animal to human tissue. This emphasizes the importance of the use of human tissue in the development of therapeutic drugs.