Safety Assessment Of Multiple Repeated Percutaneous Punctures For The Collection Of Cerebrospinal Fluid In Rats

The objective of this study was to examine the safety of multiple repeated percutaneous punctures of cisterna magna for collecting cerebrospinal fluid (CSF) and preliminarily determine the optimal time interval and volume at each collection. Sixty Wistar rats were randomly assigned to six groups: 10 d-0 mu L, 10 d-100 mu L (100 mL CSF collected at an interval of 10 days), 10 d-150 mu L, 15 d-0 mu L, 15 d-100 mu L, and 15 d-150 mu L. CSF was collected by percutaneous puncture of the cisterna magna at four time-points. Simultaneously, locomotor activity, cisterna magna pressure, and acetylcholine levels in the CSF were monitored. Compared with the 10 d-0 mu L group, the escape latency by Morris water maze was significantly prolonged in the 10 d-100 mu L and 10 d-150 mu L groups (P<0.05). Compared with the 15 d-0 mu L group, the indices of 15 d-100 mu L and 15 d-150 mu L groups had no significant differences. When compared with that at the first training, the exception of the 10 d-150 mu L and 15 d-150 mu L groups, significant differences in escape latency were found at the 6th attempt (P<0.05). Compared with baseline readings for each group, the cisterna magna pressure in the 10 d-150 mu L group began to decrease significantly from the third measurement (P<0.05). The optimal time interval during four CSF collections (100 mu L per collection) via cisterna magna percutaneous puncture was determined to be 15 days. The procedure did not significantly affect learning processes, performance, or other related indices.