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 carbachol-induced 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-(3)H]scopolamine ([(3)H]NMS) binding. The TGF-beta1 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 [(3)H]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-beta1 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 muM (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.
Akiho
Hirotada
H
Department of Medicine and Bioregulatory Science, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. akiho@intmed3.med.kyushu-u.ac.jp
Khan
Waliul I
WI
Al-Kaabi
Atheer
A
Blennerhassett
Patricia
P
Deng
Yikang
Y
Collins
Stephen M
SM
D000818:Animals; D002217:Carbachol / Q000494:pharmacology; D007371:Interferon-gamma / Q000494:pharmacology*; D007583:Jejunum / Q000166:cytology / Q000502:physiology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009119:Muscle Contraction / Q000187:drug effects*; D019832:N-Methylscopolamine / Q000378:metabolism; D017471:Receptors, Interferon / Q000096:biosynthesis; D011976:Receptors, Muscarinic / Q000187:drug effects* / Q000502:physiology*; D018125:Receptors, Transforming Growth Factor beta / Q000096:biosynthesis; D050800:STAT6 Transcription Factor / Q000172:deficiency; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D017160:Trichinella spiralis; D014235:Trichinellosis / Q000503:physiopathology; D000096743:Interferon gamma Receptor
D002217:Carbachol; D007371:Interferon-gamma; D019832:N-Methylscopolamine; D017471:Receptors, Interferon; D011976:Receptors, Muscarinic; D018125:Receptors, Transforming Growth Factor beta; D050800:STAT6 Transcription Factor; D016212:Transforming Growth Factor beta; D000096743:Interferon gamma Receptor; C494100:Stat6 protein, mouse