Experimental Study On The Concentration Of Free Ca²⁺ And The Detection Of ATP Content And ATP Enzymethe Activity In The Rat Spinal Synapse

Objective:Using a fluoroscope to observe the morphology of rat spinal synaptosomes,By measuring the ATP content and ATPase activity in the synaptophysin of the spinal cord and measuring the change of free Ca²⁺concentration in the synaptosomes of the spinal cord under normal conditions and high potassium stimulation,In order to detect the physiological activity of the prepared spinal synaptosomes,In preparation for the study and treatment of spinal cord related diseases.Methods:After preparing rat spinal synapses,Synaptosomal ATP content and ATPase activity were measured using a UV spectrophotometer.Fluorescence spectrophotometer and flow cytometer FCM were used to detect the concentration of free Ca²⁺in synaptosomes of rat synaptosomes under resting and hyperkalemia conditions.Results:1.The morphology and morphology of synaptosome in rat spinal cord were observed under electron microscope.2.The activities of ATP and ATPase were detected in synaptophysin of rat spinal cord.The activities of sodium,potassium and ATPase were higher than those of calcium and magnesium ATPase.The difference was statistically significant（P<0.01）.3.With fluorescence spectrophotometry,calcium ion concentration in synaptophysin of rat spinal cord could be detected in saline group and potassium chloride group,and they could be detected separately.The Fmin,Fmax and F values of rat spinal synaptic calcium ion concentration in the potassium chloride group were higher than that in the saline group.The difference between the two groups was statistically significant（P<0.01）;flow cytometry method was used.Fluorescence values were detected in the saline group and the potassium chloride group.The fluorescence value of the potassium chloride group was higher than that of the saline group.The difference between the two groups was statistically significant（P<0.01）.The method has a consistent change of calcium ion concentration under different conditions.Conclusion:The rat spinal synapse contains ATP,sodium and potassium ATPase,and calcium and magnesium ATPase.The calcium ions in the synaptosome of the spinal cord are hyperstimulated and the calcium ions infiltrate.The rat spinal cord synapse,which has been prepared in our country,is structurally complete and has a certain degree of The physiological activity.