Cytokine modulation of muscarinic receptors in the murine intestine

The extent to which gut motility and smooth muscle contractility are altered by intestinal inflammation depends on the nature of the underlying immune activation. The muscarinic receptor on smooth muscle plays a critical role in mediating acetylcholine- driven motor function. We examined the ability of cytokines to influence muscarinic receptor characteristics on intestinal longitudinal muscle and related the findings to studies on carbacholinduced contraction. Cells were isolated from longitudinal muscle myenteric plexus ( LMMP). Cytokine receptor expression, muscle contractility, and muscarinic agonist receptor characteristics were examined by agonist displacement of [ N- methyl-H-3] scopolamine ([H-3] NMS) binding. The TGF-beta 1 receptor ( 543 bp) and the IFN-gamma receptor 1 ( 660 bp) were identified on smooth muscle cells. Scatchard analysis revealed dissociation constant and maximum binding values for [H-3] NMS of 2.6 nM and 2.4 x 10(4) sites/ cell, respectively, in control cells. Nematode infection was accompanied by a reduction in inhibitory constant of the high-affinity sites ( K-H), and this was independent of signal transduction and activator of transcription 6. Preincubation with TGF-beta 1 enhanced longitudinal muscle contractility and decreased the K-H to 2.2 pM ( increased muscarinic receptor affinity), whereas preincubation with IFN-gamma increased the K-H to 0.4 mu M ( decreased muscarinic receptor affinity) and decreased longitudinal muscle contractility. Preincubation of LMMP with IL- 13 decreased the K-H to 0.2 nM. Cytokines exert differential effects on the muscarinic receptor on intestinal longitudinal smooth muscle. These findings explain the basis for altered muscle contractility observed in Th1 and Th2 models of inflammation, as well as in the post- nematode- infected state.