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Role of Cholinergic Signal Mediated by Muscarinic Receptors on ICCs in Smooth Muscle Spontaneous Contraction of Guinea Pig Ureter

Author: GeLiang
Tutor: LiWeiBing
School: Third Military Medical University
Course: Surgery
Keywords: Ureter Interstitial cells of Cajal Smooth muscle Voluntary contraction Muscarinic receptors
CLC: R339.2
Type: Master's thesis
Year: 2010
Downloads: 82
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Abstract


Background and Purpose: Cajal interstitial cells of the original is considered to exist only in a special type of cells of the gastrointestinal tract, gastrointestinal tract plays pacing slow wave transfer electrical signals between the adjustment signal mediated by intestinal nerves and smooth muscle role is closely related with the pathological state of a variety of gastrointestinal motility disorders (such as constipation, pseudo-intestinal obstruction), an important factor in gastrointestinal smooth muscle rhythmic contractions of functional regulation. Later, other organizations outside of the gastrointestinal tract, found the same as the immunological characteristics of the interstitial cells of Cajal of the gastrointestinal tract, cells with similar morphological characteristics, previous known as ICC-like cells (ICC-like cells) have been collectively referred to as interstitial cells of Cajal (ICCs). Physiological circumstances, ureteral smooth muscle very similar to the gastrointestinal tract, that existence is not dependent on nerve spontaneous peristaltic, although the exact mechanism has not been elucidated. Existing research found that there are ICCs in guinea pig and human urinary tract. Reported in the literature than normal children and children with congenital ureteropelvic obstruction of the renal pelvis and ureter ICCs significantly reduced, prompted the ICCs and ureter excited origin and regulation are closely related. Further research ICCs voluntary contraction in the ureter in its response to the neurotransmitter, helps to clarify the effect and mechanism of ICCs in the regulation of ureteral smooth muscle function, and provide a new cut for the treatment of diseases of upper urinary tract dysfunction point. This issue through muscle strips of guinea pig ureter muscle strips vitro experimental observation, observe the muscle strips nonspecific M cholinergic receptor agonist carbachol voluntary contraction before and after the change, and ICCs function in specific blocker voluntary contraction and carbachol response to changes in addition to the application of double immunofluorescence staining ureter ICCs M receptor subtype expression so as to clarify the ICCs on M receptor-mediated cholinergic signal ureteral smooth muscle voluntary contraction provide new experimental evidence. Methods: Experimental animals were clean healthy Hartley guinea pigs. (1) carbachol ureteral smooth muscle voluntary contraction and ICCs impact: 20 guinea pigs were randomly divided into normal control group, the blocking ICCs group, n = 10. Blocking the ICCs group muscle strips balance prior to testing and detection are placed containing a final concentration of 10-4mol / L of Glivec of the Kreb's solution, normal control group placed without Glivec Kreb's solution. Take guinea pigs the left upper ureteral 1.0cm organization made vitro smooth muscle strips detected in the final concentration of 10-4mol / L carbachol role before and after the voluntary contraction frequency and amplitude. (2) ureteral ICCs M receptor subtype expression: c-kit M1 ~ M5 Immunofluorescence double staining assay on the guinea pig ureter ICCs M receptor subtype expression. Results: (1) under certain pre-load (2.0g) was added 1mol / L KCl solution 200μL ureteral smooth muscle strips normal control group of guinea pigs in vitro can be induced by a voluntary contraction frequency (3.4 ± 0.84) beats / min, volatility (0.105 ± 0.0228) g. (2) stable voluntary contraction in muscle strips, added 0.01mol / L carbachol 100μL, muscle strips voluntary contraction frequency (4.9 ± 0.88) beats / min, more dosing significantly increased faster (p lt; 0.01) ; contraction amplitude (0.106 ± 0.0210) g, compared to dosing, no significant change (p gt; 0.05). (3) 30 minutes with a 10-4mol / L Glivec balance, a specific inhibitor of the ICCs same method detected the blocking the ICCs group ureteral muscle strips without voluntary contraction, added 0.01mol / L Kabbah 100μL still no reaction. (4) Immunofluorescence double staining found on the guinea pig ureter ICCs express both M2, M3 M receptor subtypes. The Conclusion: ICCs are likely excited about the origins of the voluntary contraction of ureteral smooth muscle and cholinergic nerve regulation by the M2 and M3 receptor subtypes accept.

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