Study On ChAT Changes Of The Spinal Cord After C₄Transferring To C₆

objectiveExperiment one: Using cholera toxin B-horseradish peroxidase （CB-HRP）injecting in C₄nerve root, trace its motor nucleus in the spinal cord ofpositioning. Experiment two:First,establishing the Wistar rat right brachial plexusroot avulsion model, and then C₄nerve root transfering to C₆repairing nerveroot,observing the right forelimb general situation, triceps brachii-twoneurophysiological, C₄nerve root corresponding segment of spinal cord anteriorhorn motor neurons acetylcholine transferase （ChAT） expression change,neurotransmitter changes reveal the reasons that small nerve alternative sciaticnerve got recover motor function.MethodsExperiment one:Getting6Wistar rats after weighing, ketamine injection onintraperitoneal injection of anesthesia, the doses is100mg/kg, then satisfactionwith anesthesia, the rat limbs and head are respectively fixed on the operationtable using a rubber band around the edge, to prevent rats in the operationprocess of moving, and make it a supine position; the neck skin hair removal,disinfection, bed sheet. takes in front of the neck incision,length is4.0cm, andincisie skin、platysma, from the right side of the sternocleidomastoid muscle andmuscle neck gap line after isolated blunt, cut off the sternocleidomastoid muscle,thorough hemostasis, resection after the show C₄nerve root, and distal to thefull free. Then use the micro syringe tip needle oblique assassination into C₄ nerve root slow infusion of8μl CB-HRP, then chase a suture, after24hours,after all animal perfused with4%neutral paraformaldehyde fixation, takingC₃^C₅segment of the spinal cord, the conventional dehydration,paraffin-embedded sections, line3,3,5,5, four methyl aniline （TMB） staining.Experiment two:30Wistar rats were randomly divided into groups,10eachgroup A,B,C. GroupA、B establish C₅^T1brachial plexus avulsion model,group Ais the experimental group, C₄transposition for repairing treatment; group B iscontrol group, in the intervertebral foramen is cut at C₄and C₆nerve root, noanastomosis; group C as normal control group, without operation. Regularobserve animal limb function of three groups. Postoperative three months,â‘ three groups of animal are right forelimb electromyography （EMG）;â‘¡group Agot nerve0.5cm between C₄and C₆anastomotic distal, group B got nerve0.5cm in the stump of C₆, group C got nerve0.5cm in C₆,were stained with HEand then observed with nerve fibers;â‘¢at the same time,got C₄spinal cordtissue of three group animal,stained with ChAT,calculated the number of ChATspinal anterior horn motor neuron immunoreactive positive neurons.（10in eachanimal） C₄segment of the spinal cord tissue, sections, stained with ChAT,testing ChAT changes.ResultsExperiment one:C₃^C₅segment of the spinal cord by TMB staining, all colored parts arebasically located in the C₄segment of the spinal cord, the blue part is shown inthe figure.Experiment two:1. Rats limb function and skin ulcers in each group: group C movement,skin were normal. The immediate postoperative A, B rats right forelimb wasstretched state, can not be off when walking, noticeable limp; postoperative oneweek, all right forelimb triceps brachii-two atrophy; postoperative40days in group A triceps brachii-two gradually returned to normal, group B is stillshrinking state; after3months right upper of group A can flex elbow, withoutobvious lameness,when walking, group B showed no significant improvement.5rats of group B at postoperative1months limb skin ulcer appeared, healing afterlocal disinfection, in the A group, no skin ulcer.2. Electromyography: Latency amplitude （AMP） and nerve conductionvelocity （NCV） of group A, C is similar basically, differences between groupswere not statistically significant （P>0.05）, B group did not detect potential.3．Nerve fiber structure in each group: After stained with HE, group Athere is a lot of regenerating nerve fibers and myelin, uniform, dense, epicardialcontinuous, nerve fiber is arranged neatly; group B can be seen scattered insmall nerve fibers, disordered disordered, see a lot of connective tissue anddifferentiated into Schwann cells of a group; group C visible myelinated nervefiber distribution uniform, dense, neat arrangement.4C₄spinal anterior horn motor neuron number of the ChATimmunoreactive respectively （23.27+3.35）,（19.53+2.97）,（25.53+3.42） in A, B,C group, A group compared with B group, P <0.05,0.01, A group and C group issimilar （P>0.05）.ConclusionsAfter C₄transfering to repair and treatment in brachial plexus root avulsionmodel, the number of ChAT spinal anterior horn motor neuron immunoreactivepositive neurons is up, which may be one of the reasons that a small nervetransposition alternative sciatic nerve gets motor function recovery.