Basic-alimentary tractMechanisms Underlying the Maintenance of Muscle Hypercontractility in a Model of Postinfective Gut Dysfunction
Section snippets
Materials
The following materials were used in this study: collagenase (CLS type l), trypsin inhibitor, BSA, phenylmethylsulfonyl fluoride (PMSF), aprotinin, leupeptine, nonidet P-40, acrolein, PGE2, peroxidase conjugated-anti-goat/sheep IgG from Sigma Chemical (St. Louis, MO); HEPES from Bioshop Canada (Burlington, ON, Canada); IL-4, IL-13, TGF-β1, TGF-β1 immunoassay kit from R&D Systems (Minneapolis, MN); DMEM, antibiotic-antimycotic from Gibco BRL Life Technologies (Gaithersburg, MD); PGE2 assay kit
PCR for IL-4R and TGF-β Type IIR
As shown Figure 1, PCR products for IL-4Rα (241 base pairs for soluble IL-4R [sIL-4R] and 127 base pairs for membrane IL-4R [mIL-4R]) and 2 distinct forms of mouse TGF-β type IIR (526 base pairs for mTGF-βRII1 and 601 base pairs for mTGF-βRII2) were expressed on primary cultured muscle cells isolated from control mice (Figure 1).
T spiralis–Induced Muscle Hypercontractility
We next examined the characteristics of single muscle cells isolated from the longitudinal muscle layer from control and T spiralis–infected mice. In muscle from
Discussion
We used isolated smooth muscle cells in this study to localize the action of mediators expressed in the model because direct effects on muscle are more difficult to demonstrate in muscle strips because these retain intrinsic innervations.21 We showed that cells isolated from the longitudinal muscle layer of T spiralis–infected mice exhibited a substantially greater degree of shortening on exposure to carbachol than cells isolated from control mice. This finding indicates that the
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Supported by a grant from the Canadian Institutes of Health Research (to S.M.C.) and from a Research Initiative Award (to H.A.) from the Canadian Association of Gastroenterology and AstraZeneca.