Stereotaxic table

Rats were prepared for intrathecal (i.t.) injection by implanting catheters according to the method of Yaksh and Rudy [39 (link)] under pentobarbital (60 mg/kg i.p.) anesthesia. The intrathecal catheter consisted of polyethylene tubing that was 12 cm long (PE 10, Intramedic; Clay Adams, Parsippany, NJ) with an outside diameter of 0.4 mm and a dead space of 10 μL that had been sterilized by immersion in 70% (v/v) ethanol and been fully flushed with sterile water before insertion. Rats were placed on a stereotaxic table (David Kopf Instruments, Tujunga, CA), and an incision was made in the atlantooccipital membrane. The catheter (7.8 cm of its length) was carefully introduced into the subarachnoid space at the rostral level of the spinal cord lumbar enlargement (L4-L5). After the implantation, the first injection of 10 μL of water was performed slowly, and the catheter was tightened. After catheter implantation, the rats were monitored for physical impairments. Those showing motor deficits (ca 5%) were excluded from further study. Animals were allowed a minimum of 1 week to recover after the surgery before the experiment began. Water for injection or respective drugs were delivered slowly (1-2 min) in a volume of 5 μL through the i.t. catheter and were followed by 10 μL of water, which flushed the catheter.

Intrathecal (ith.) administration of substances was achieved by implanting catheters according to the method described earlier (Yaksh and Rudy 1976 (link)). The procedure was performed under pentobarbital (60 mg/kg; i.p.) anaesthesia. The polyethylene catheters (PE 10, Intramedic; Clay Adams, Parsippany, NJ) were sterilized before the operation in 70% (v/v) EtOH and flushed with water for the injection directly before insertion. Rats were placed on a stereotaxic table (David Kopf Instruments, Tujunga, CA), and an incision in the atlantooccipital membrane was made for the introduction of the catheter (7.8 cm long) into the subarachnoid space. After insertion, the catheters were slowly flushed with 10 µL of water for the injection, and the tip was tightened for separation from the environment. After implantation, the rats were observed for physical impairments and allowed to recover for a minimum of 1 week before the actual experiment.

Rats were prepared for intrathecal (i.t.) injection by implanting catheters under pentobarbital anaesthesia (60 mg/kg i.p.). The intrathecal catheter consisted of polyethylene tubing that was 12 cm long (PE 10, Intramedic; Clay Adams, Parsippany, NJ) with an outside diameter of 0.4 mm and a dead space of 10 µl that had been sterilised by immersion in 70% (v/v) ethanol and fully flushed with sterile water before insertion. Rats were placed on a stereotaxic table (David Kopf Instruments, Tujunga, CA), and an incision was made in the atlanto-occipital membrane. The catheter (7.8 cm of its length) was carefully introduced into the subarachnoid space at the rostral level of the spinal cord lumbar enlargement (L4–L5) according to the method of Yaksh and Rudy [53] (link). After the implantation, the first injection of 10 µl of water was performed slowly and the catheter was tightened. One day after catheter implantation, the rats were monitored for physical impairments. Those showing motor deficits were excluded from further study. Animals were allowed a minimum 1 week of recovery after the surgery before the experiment began. Water for injection or respective drugs were delivered slowly (1–2 min) in a volume of 5 µl through the i.t. catheter and were followed by 10 µl of water for injection, which flushed the catheter.