OBJECTIVE: In humans, fetal in utero meconium (MEC) passage rarely occurs before term gestation. We hypothesized the existence of inhibitory mechanism(s) preventing colonic motility and MEC passage prior to term. STUDY DESIGN: Longitudinal smooth muscle strips prepared from distal colon of preterm ovine fetuses (130-132 d; term = 148-152 d) were examined for their contractile responses to muscarinic receptor agonist (bethanechol) and both nonspecific (atropine) and receptor subtype specific antagonists (M1: pirenzepine dihydrochloride, M2 methoctramine, M3: 4-diphenylacetoxy-N-methlpiperidine methiodide [4-DAMP] and M4: tropicamide) in an in vitro organ bath system. Effects of corticotrophin releasing factor (CRF) and Urocortin I (URO-I), known modulators of colonic motility and smooth muscle contractility, were studied on bethanechol-induced contractility. Immunohistochemical analysis was performed to confirm the expression of CRF and URO-I, and muscarinic and CRF R2 receptors in distal colon. RESULTS: Bethanechol induced smooth muscle contractions via muscarinic receptor subtype M3. CRF and URO-I elicited a significant inhibition of bethanechol induced contraction. Immunohistochemical analysis verified the expression of muscarinic receptor subtype M3, CRF, URO-I and CRF-receptor-R2 in distal colon. CONCLUSION: Inhibition of M3 dependent distal colonic motility by CRF system may prevent the passage of MEC in the preterm ovine fetus.
OBJECTIVE: In humans, fetal in utero meconium (MEC) passage rarely occurs before term gestation. We hypothesized the existence of inhibitory mechanism(s) preventing colonic motility and MEC passage prior to term. STUDY DESIGN: Longitudinal smooth muscle strips prepared from distal colon of preterm ovine fetuses (130-132 d; term = 148-152 d) were examined for their contractile responses to muscarinic receptor agonist (bethanechol) and both nonspecific (atropine) and receptor subtype specific antagonists (M1: pirenzepine dihydrochloride, M2 methoctramine, M3: 4-diphenylacetoxy-N-methlpiperidine methiodide [4-DAMP] and M4: tropicamide) in an in vitro organ bath system. Effects of corticotrophin releasing factor (CRF) and Urocortin I (URO-I), known modulators of colonic motility and smooth muscle contractility, were studied on bethanechol-induced contractility. Immunohistochemical analysis was performed to confirm the expression of CRF and URO-I, and muscarinic and CRF R2 receptors in distal colon. RESULTS:Bethanechol induced smooth muscle contractions via muscarinic receptor subtype M3. CRF and URO-I elicited a significant inhibition of bethanechol induced contraction. Immunohistochemical analysis verified the expression of muscarinic receptor subtype M3, CRF, URO-I and CRF-receptor-R2 in distal colon. CONCLUSION: Inhibition of M3 dependent distal colonic motility by CRF system may prevent the passage of MEC in the preterm ovine fetus.