ABSTRACT
Muscle-specific tyrosine kinase (MuSK) is essential for clustering of acetylcholine receptors (AChRs) at embryogenesis and likely also important for maintaining synaptic structure in adult muscle. In 5 to 7% of myasthenia gravis (MG) cases, the patients' blood contains antibodies to MuSK. To investigate the effect of MuSK-MG antibody on synapse regeneration, notexin was used to induce damage to the flexor digitorum brevis muscle. We administered aliquots of MuSK-MG patients' plasma to the flexor digitorum brevis twice daily for a period up to 21 days, and muscles were investigated ex vivo in contraction experiments. AChR levels were measured with (125)I-alpha-bungarotoxin, and endplates were studied with quantitative immunohistochemistry. In normal muscles and in 14-day regenerated muscles, MuSK plasma caused impairment of nerve stimulus-induced contraction in the presence of 0.35 and 0.5 mmol/L Ca(2+) with or without 100 to 400 nmol/L tubocurarine. Endplate size was decreased in regenerated muscles relative to controls; however, we did not observe such differences in muscle not treated with notexin. MuSK plasma had no effect on the amount and turnover rate of AChRs. Our results suggest that anti-MuSK antibodies influence the activity of MuSK molecules without reducing their number, thereby diminishing the size of the endplate and affecting the functioning of AChRs.
Subject(s)
Motor Endplate/physiopathology , Myasthenia Gravis/blood , Myasthenia Gravis/enzymology , Receptor Protein-Tyrosine Kinases/blood , Receptors, Cholinergic/blood , Regeneration/physiology , Animals , Calcium/pharmacology , Elapid Venoms/pharmacology , Female , Immunohistochemistry , Mice , Models, Biological , Motor Endplate/drug effects , Muscle Contraction/drug effects , Myasthenia Gravis/physiopathology , Neuromuscular Junction/drug effects , Neuromuscular Junction/pathology , Organ Specificity/drug effects , Plasma , Receptors, Cholinergic/metabolism , Regeneration/drug effects , Tubocurarine/pharmacologyABSTRACT
BACKGROUND: Besides regulation of upper gastrointestinal motility, motilin seems to play a role in the inflammatory response. Motilin receptor expression in human intestine has not been studied thoroughly. This study aimed to describe the intestinal distribution of motilin receptors in inflammatory bowel disease (IBD) and control patients. METHODS: Quantitative autoradiography, immunohistochemistry, and reverse-transcriptase polymerase chain reaction (RT-PCR) were used to detect motilin receptors in tissue of 25 IBD patients (13 Crohn's disease [CD], 12 ulcerative colitis [UC]) and 19 patients with a neoplasm (controls). RESULTS: Median muscular motilin binding was 3 and 8 fmol/g tissue in colon and ileum, respectively. In the gastroduodenal region the median was higher (93 fmol/g). In UC colonic muscular motilin binding was significantly increased compared to controls (7 vs. 3 fmol/g, P < or = 0.05). Expression in CD was similar to controls. Besides the binding found in the muscular compartment, motilin binding was also found in the mucosa, which was even higher than in the muscle (3 versus 11 and 8 versus 27 fmol/g for colon and ileum (P < or = 0.06), respectively). RT-PCR and immunohistochemistry confirmed the mucosal motilin receptor expression. The mucosal motilin receptors were located in the epithelial cells. In the muscular compartment receptors were strongly present in the myenteric plexus and weakly in the smooth muscle cells. In IBD tissue the expression pattern was not different. CONCLUSIONS: The motilin receptor is expressed in human colonic and ileal smooth muscle. Further, motilin receptor expression was also shown in the mucosa. Muscular binding in UC patients is increased but no different expression pattern was found.
