Abstract
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 μL, 10 d-100 μL (100 μL CSF collected at an interval of 10 days), 10 d-150 μL, 15 d-0 μL, 15 d-100 μL, and 15 d-150 μL. 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 μL group, the escape latency by Morris water maze was significantly prolonged in the 10 d-100 μL and 10 d-150 μL groups (P<0.05). Compared with the 15 d-0 μL group, the indices of 15 d-100 μL and 15 d-150 μL groups had no significant differences. When compared with that at the first training, the exception of the 10 d-150 μL and 15 d-150 μL 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 μL group began to decrease significantly from the third measurement (P<0.05). The optimal time interval during four CSF collections (100 μL 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.
Highlights
In animal experiments studying the central nervous system (CNS), cerebrospinal fluid (CSF) samples are often needed to monitor chemical analytes including broad biological markers or neuro-transmitters (1–3)
The rat stereotaxic apparatus was a product of BIOPAC (USA); the Morris water maze was purchased from Huaibei Zhenghua Biologic Apparatus Facilities Co., Ltd. (China); the open field test cage was provided by Huaibei Zhenghua Biological Instrument Equipment Co., Ltd.; the ACQUITY UPLC-I-CLASS was purchased from the Waters Company (USA); the XevoTQD-triple quadrupole mass spectrometer was a product from Waters Company; the MS105 type 1/100,000 analytical balance was purchased from the METTLER TOLEDO Company (Germany); the BL-420 biological function experimental system was provided by Chengdu Taimeng Technology Co., Ltd. (China); and the moorVMS dual-channel laser Doppler flowmeter was purchased from Moor Instruments (UK)
The results of preliminary experiments showed that a collection volume of less than 150 mL did not cause blood contamination, it is unknown whether a collection volume of 150 mL has any effect on neurological function
Summary
In animal experiments studying the central nervous system (CNS), cerebrospinal fluid (CSF) samples are often needed to monitor chemical analytes including broad biological markers or neuro-transmitters (1–3). Acetylcholine is an important neurotransmitter in the central nervous system. It plays a special role in cognitive functions such as learning and memory. The main pathway of learning and memory is composed of central cholinergic transmission (4–7). Most researchers adopt microdialysis and transdural puncture of the cisterna magna under direct vision for multiple repeated collections of CSF. Microdialysis for multiple repeated collections of CSF is safer, but the recovery is low, which is not conducive to the detection of metabolites of some micro-components
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