Comparison Among Different Approaches for Sampling Cerebrospinal Fluid in Rats
Section snippets
INTRODUCTION
Time-resolved sampling of cerebrospinal fluid (CSF) enables evaluation of dynamic biochemical and pharmacological events in the extracellular milieu of the central nervous system (CNS) 1, 2, 3, 4, 5, 7, 9, 10, 11, 12, 18, 19, 20, 21, 23. The preferred origin of CSF in rats is the cisterna magna. It is readily available and has a large volume (≈ 190 μl; cistern and adjacent subarachnoid spaces ≈ 246 μl; one lateral ventricle just ≈ 10 μl) [3]. The techniques in use require surgical insertion of
Animals
Adult outbred Sprague-Dawley rats were used (n = 360; body weight ≈ 300 g; B and K Labs. AB, Stockholm, Sweden). The light was on between 6 a.m. and 6 p.m. The animals had free access to pelleted food and water. Permission to the experiments was granted by the Animal Experiments Ethical Committee (authorizations O , O , and A ).
Anesthesia and Surgery
One group of animals was anaesthetized during the surgery and the CSF sampling either by intraperitoneal injection of a mixture of pentobarbital sodium (60
General Comments
Both surgical procedures could be completed by the surgeon in 1 h, including the time required for the polymerization of the acrylic cement. Based on the records for 193 rats, ≥ 65% of the tubes implanted according to surgical technique I were patent after one day. Surgical techniques II had a patency rate ≥ 90% after one day (n = 128).
The tightness of the tube was checked by dissection of the extracranial tissue 2 h after the surgery in animals with EBA infused into the cisterna magna at the
DISCUSSION
A major problem in experiments with time-resolved sampling of CSF is the damage caused by the surgery necessary for the implantation of the sampling tube. The commonly used technique, surgical technique I, invariably damaged the meaninges and the adjacent nervous tissue. These results are in agreement with those published previously 4, 7, 9, 19, 20, 21, 23. In contrast, surgical technique II caused negligible evidence of trauma, restricted to the immediate vicinity of the tube and hole, or none
Acknowledgements
We thank Mr. K. Hansson, Mr. T. Jönsson, Ms. I.-L. Larsson, Ms. E. Malm, Ms. E. Mattsson, Ms. M. Miikiver, and Mr. M. Vona for their expert assistance and Mr. T. Seeman, MD, PhD, for valuable help.
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2012, Journal of Neuroscience MethodsCitation Excerpt :Since CSF drawing is very frequently used procedure, what is the best method has raised our concern. In this study, we performed and compared the three common ways: skull drilling (Huang et al., 1996), lumbar puncture (Wang et al., 2005; De la Calle and Paíno, 2002) and atlanto-occipital membrane puncture (Haslberger and Gaab, 1986; Vistelle et al., 1989; Huang et al., 1996), in order to find an ideal one that is the simplest, fastest, and with the highest repeatability. Human lumbar puncture needles (#4, Bangcheng Co., Shanghai, China) with about 0.4 mm outside diameter: they were cut to be 4 cm in length and sharpened under the help of microscope with a grindstone (#1200), the tips were made less sharp than the original ones (see the part of discussion and Fig. 3); crew cut bone drills (RA-II, Yancheng Co., China, 8 mm outside diameter): to avoid easily break the cerebral dura mater, the tip of the drill were blunted a little using #200 first and then #1200 grindstones; silicone tubes with 1.5 mm inside diameter and 2 mm outside diameter; suture needles; sutures; alcohol swabs; dry cotton balls; scissors, forceps; animal fixing frame; 1 ml, 5 ml and 20 ml syringes; #5 and #7 syringe needles; trays; 1% of pentobarbital sodium.
Cisterna magna cannulated repeated CSF sampling rat model - effects of a gamma-secretase inhibitor on Aβ levels
2012, Journal of Neuroscience MethodsCitation Excerpt :The transcutaneous approach includes both direct needle access (Takasugi et al., 2005; Nirogi et al., 2010) or open surgical access to the cisterna magna in the back of the neck (Pegg et al., 2010). In contrast, surgical trans-occipital bone approach (Sarna et al., 1983; Consiglio and Lucion, 2000) or catheterization via the atlanto-occipital membrane in the back of the neck (Huang et al., 1996) has been used to permanently implant catheters into the cisterna magna to enable repeated CSF sampling. Some reports have also used an indwelling cannula in the lateral ventricle to sample CSF over time via parietal bone access (Cassar et al., 2010).
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2010, Journal of Neuroscience MethodsCitation Excerpt :Overall, these results suggest that levels of some biomarkers may be altered in CSF from cannulated versus non-cannulated rats. In addition to the two collection methods examined here, alternative techniques, cannulated and non-cannulated, are used by others (Huang et al., 1996; Pegg et al., 2010). Each method of collection may affect CSF biomarkers distinctly.