【Abstract】Objective To establish a new catheterization of the spinal subarachnoid space in mouse and to compare with the traditional catheterization, which can be used for the research on neurophysiology and pain treatment. Methods forty male Kunming mice weighing 24-28g were randomly divided into two groups: Wu group and axis-atlantoaxial group (AA group), respectively catheter from the intervertebral space of lumbar 5-6 and atlantoaxial to the subarachnoid space. After the models were established, we observed the percentage of death and paralysis of limbs, and also measured 50%paw withdrawal threshold (50%PWT), tail flick latency and analgesic effect of morphine to compare the difference of two groups. Results In the case of the same death rate in Wu group and AA group, the percentage of paralysis of limbs in AA group was obviously lower than Wu group. Within 7 days of catheterization, 50%PWT and the TF of AA group was closest to baseline. Morphine analgesic experiment showed the new method adapt to long-term injection. Conclusion The atlantoaxial catheterization model of the spinal subarachnoid space is characterized by its faster recovery and less injury, which was a effective intrathecal catheterization method. 【Key words】Intrathecal Catheterization, Atlas And Atlantoaxial, Animal Model