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. 2009 Mar 30;178(1):116-9.
doi: 10.1016/j.jneumeth.2008.12.001. Epub 2008 Dec 6.

A simple and rapid method to collect the cerebrospinal fluid of rats and its application for the assessment of drug penetration into the central nervous system

Ramakrishna Nirogi  1 , Vishwottam Kandikere, Koteshwara Mudigonda, Gopinadh Bhyrapuneni, Nageswararao Muddana, Ramanatha Saralaya, Vijay Benade
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A simple and rapid method to collect the cerebrospinal fluid of rats and its application for the assessment of drug penetration into the central nervous system

Ramakrishna Nirogi et al. J Neurosci Methods. .

Abstract

Many central nervous system (CNS) drug discovery programs require the successful collection of cerebrospinal fluid (CSF) for assessing CNS penetration and distribution of new chemical entities. The objective of the present investigation was to simplify the technique for collecting maximum CSF from cisterna magna of the rats. Rat was anesthetized with 5% halothane and positioned in a stereotaxic frame. The rat head was flexed downward at approximately 45 degrees , a depressible surface with the appearance of a rhomb between occipital protuberances and the spine of the atlas becomes visible. The 23 G needle was punctured into the cisterna magna for CSF collection without making any incision at this region. The blunt end of the needle was inserted into a 10 in. length of PE-50 tubing and other end of the tubing was connected to a collection syringe. The non-contaminated sample was drawn into the syringe by simple aspiration. This technique is simple and can be performed by one person. The technique has a greater than 95% success rate of CSF collection and it was free of red blood cell contamination. In addition, it yielded 100-120 microL of CSF per rat. This method is simple, effective, and easy to perform and has been successfully applied in preclinical screening of novel chemical entities in neuropharmacotherapy for CNS use. The present method is demonstrated by studying the CSF concentrations of carbamazepine and raclopride.

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