Use of the Morris Water Maze and Acoustic Startle Chamber to Evaluate Neurologic Injury after Asphyxiai Arrest in Rats

Abstract

The study was peformed to assess the utility of the Morris water maze (MWM) and acoustic startle reflex (ASR) for evaluating neurologic outcome in a rat model of asphyxial cardiac arrest. Rats were anesthetized, intubated, and chemically paralyzed. Control animals were decannulated and, after awakening, were extubated and returned to their housing. Experimental animals were asphyxiated by disconnecting the ventilator. Approximately 3.5 min after the disconnection, there was no measurable pulse. After 7 min of asphyxia, they were then resuscitated with resumed ventilation, chest compressions, epinephrine, and sodium bicarbonate. All animals were assigned to either MWM or ASR testing. The MWM is a 6-ft diameter tank filled with opaque water. In a fixed location of the tank, a 4-inch diameter escape platform is submerged just below the surface. MWM animals were tested on post-injury d 16-21 by recording the path and time taken to escape from three randomly assigned locations per d. ASR animals had s.c. leads placed over the right triceps and tibialis anterior muscles. The latency and rectified amplitude of the ASR was measured by recording the electromyographic impulse generated when the animal was startled by an acoustic stimulus. Animals were tested on post-injury d 6 and 7. After the last test session for each group, the animals' brains were removed for histopathologic examination. Asphyxiated MWM animals took longer to find the platform, and their paths were less direct than control animals (analysis of variance p < 0.05). The ASR of asphyxiated ASR animals had greater amplitude and shorter latency compared with controls (analysis of variance p < 0.05). Histologic examination revealed no abnormalities in control animals, but 80% of asphyxiated brains showed hippocampal neuronal injury and/or reactive gliosis in the CA1 segment. Abnormalities were more commonly detected in animals killed 7 d post-injury (ASR protocol) compared with animals killed 21 d post-injury (MWM protocol). We conclude that the MWM and ASR are useful for detecting neuronal injury in asphyxiated rats.