Study About The Distribution Of Connexin 40,43,45 In Different Parts Of Atrium In Dogs

ObjectiveAtrial arrhythmia is very common clinically, seriously do harm to people' s health. Most atrial arrhythmias are caused by reentry, reentry was concerned with electrical conduction functional longitudinal dissociation .The development of RF provid the most effective therapy of Atrial arrhythmia. Meanwhile it is clear that atrial arrhythmia is not only functional,but also has anatomic foundation. Up to now, the morphological foundation of atrial arrhythmia is still disputable. The electrical activity and rhythmic contraction of normal heart is base on the integrity of information passage transfer between cardiac muscle cell. Cell junction communication is one of the important ways of intercellular communication in animal body. The architectonic foundation of information transfer between cells is cell gap junction .The important architectonic reconstituent of cell gap junction is connexin(Cx) .Cx is important in cardiac muscle communications, electrical signal conduction and normal rhythmical contract .Recently there are some experiment about gap junction(GJ) and arrhythmia (ARR). Decrease of GJ will cause the action potential conduction of velocity step down,make reentry easily happen,then cause ARR. But there are not experiment about that are there any diferences about the distribution of connexin 40, 43 and connexin 45 in atrium This experiment discuss how atrium and cell of CSO does in arrhythmia.By studying the cell of atrium and CSO form courser and the characteristic of Cx disposition.Materials and Methods1 ImmunofluorescenceAdult 10 dogs,abdominal cavity anesthetize. Take the whole heart,take auricle of right atrium,the smooth part of right atrium,CSO,make frozen section as rule,fix it in formaldehyde solution with bitter acid (2% formaldehyde +0.2% bitter acid) for 45 min,rinse it for 3 times by PBS,each time for 5 min,incubate it over night by 1: 80 Cx40 43 45 polyclonal antibody .The next day rinse it for 3 times by PBS,each time for 5 min. Incubate it in room temperature for 0.75h by 1: 100 antibody signed by FITC. Rinse it for 3 times by PBS,each time for 5 min,mount it by 60 % glycerine. The whole process protect it from light2 Confocal microscopyObserve by Confocal microscopy,in each slide we take 4-6 field of vision. Store digitization image in PC.We use a soft named Image Analysis for image analysis. Determin the image element density of Cx40 Cx43 and Cx45,meanwhile determine fluorescence light spot size of Cx40 Cx43 and Cx45,take image element density and fluorescence light spot size as amount of Cx40 Cx43 and Cx45 .Carry out quantitive analysis.ResultImage Analysis show that the image element density of Cx43 in the smooth part of right atrium and CSO decrease 22.2 % and 40.0 % compared with auricle of right atrium, P< 0. 01. The image element density of Cx40 in the smooth part of right atrium and CSO decrease 29.9 % and 37.1 % compared with auricle of right atrium, P< 0. 01. We did not see Cx45 express in auricle of right atrium , the smooth part of right atrium and CSO.Conclusion1 We found that the distribution of Cx 40, 43 in atrium is abundant, We did not see Cx45 express in atrium muscle.2 We found that the distribution of connexin 40, 43 in atrium is different. 3 We can find out that the disposition of Cx40 Cx43 in auricle of right atrium >that of the smooth part of right atrium>that of CSO.the distribution has significant difference.