Lumbar intrathecal catheterization in rats targeting the cerebral cortex: a drug delivery method and validation

Intrathecal (IT) drug delivery is a critical technique for bypassing the blood-brain and blood-spinal cord barriers in preclinical CNS research. However, conventional rat catheterization methods suffer from high rates of neurologic complications, poor reliability, and unverified dosing due to epidural reflux and inconsistent supraspinal distribution. Our objective was to develop and validate an improved method for lumbar IT catheterization in rats that ensures distribution to the brain and to confirm supraspinal pharmacodynamic target engagement. We describe a refined microsurgical technique using dural puncture under direct visual control at the L6-S1 interlaminar space, a site chosen for its anatomical safety margin. The method uses a small-bore (0.33 mm OD) polyurethane (PU) catheter to minimize durotomy size, air-bubble tracking to compensate for catheter dead volume, and epidural sealing with Surgifoam^® to minimize reflux. Visualization using Evans Blue dye confirmed complete neuraxial distribution from a single 30 μL lumbar bolus injection, with dye reaching the ventral/dorsal brain cisterns. Pharmacodynamic validation of cortical exposure was achieved using an S6 kinase 1 (S6K1) inhibitor. Lumbar IT administration over a period of 6 hours via a pump resulted in significant supraspinal S6K1 engagement, demonstrated by a reproducible reduction in S6 phosphorylation in the cerebral cortex.