ICV Cannulation

See also Amygdala for other cannulation coordinates.

Lateral Ventricular Cannula:

1.2 mm caudal to bregma

1.5 mm lateral to the midline

4 mm below the skull surface

Method

Each rat was stereotaxically implanted with a 22 gauge, stainless steel, guide cannula (Plastics One, Roanoke, VA) aimed towards the lateral cerebral ventricle (1.2 mm caudal to bregma, 1.5 mm lateral to the midline, and 4 mm below the skull surface). Guide cannulas were held in place with dental acrylic bonded to stainless steel screws anchored to the skull. An obdurator was inserted into each guide cannula and remained in place except during injections when it was removed and replaced with an injector that extended 1.0 mm beyond the tip of the guide cannula.

Test

To test the patency and placement of the cannula in the lateral ventricle, water-replete rats were injected with 100 ng human angiotensin II (Ang II; Sigma Chemical Co, St Louis, MO) dissolved in 5 µl volume of saline (0.15 M). The volume of all icv injections was 5 µl, delivered over 30s with a handheld 50 µl syringe (Hamilton Co, Reno, NV). Immediately after the ICV injection, rats were returned to their home cage with access to a water bottle. The latency to drink was recorded; rats that failed to drink within 2 minutes of the angiotensin II injection were dropped from the study. Cannula placements were also verified postmortem by sectioning through the brain. ICV injections of 5 µl volume were given by hand to lightly restrained rats using a 50 µl Hamilton microsyringe.

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