Increased anesthesia time using 2,2,2-tribromoethanol–chloral hydrate with low impact on mouse psychoacoustics

• Combination avertin–chloral hydrate increases anesthesia in mice to 1 h.
• Avertin–chloral hydrate anesthesia slightly suppresses ABR Wave I, II and V amplitudes.
• Avertin–chloral hydrate has minimal effects on ABR Wave I, III and V latencies.
• Avertin–chloral hydrate causes mild intestinal vasodilation.
• Avertin–chloral hydrate has low impact on mouse health in the post-anesthetic period.

BackgroundTo examine psychoacoustics in mice, we have used 2,2,2-tribromoethanol anesthesia in multiple studies. We find this drug is fast-acting and yields consistent results, providing 25–30 min of anesthesia. Our recent studies in binaural hearing prompted development of a regimen to anesthesia time to 1 h. We tested a novel cocktail using 2,2,2-tribromoethanol coupled with low dose chloral hydrate to extend the effective anesthesia time.New methodWe have established an intraperitoneal dosing regimen for 2,2,2-tribromoethanol–chloral hydrate anesthesia. To measure efficacy of the drug cocktail, we measured auditory brainstem responses (ABRs) at 10 min intervals to determine the effects on hearing thresholds and wave amplitudes and latencies.ResultsThis novel drug combination increases effective anesthesia to 1 h. ABR Wave I amplitudes, but not latencies, are marginally suppressed. Additionally, amplitudes of the centrally derived Waves III and V show significant inter-animal variability that is independent of stimulus intensity. These data argue against the systematic suppression of ABRs by the drug cocktail.Comparison with existing methodsUsing 2,2,2-tribromoethanol–chloral hydrate combination in psychoacoustic studies has several advantages over other drug cocktails, the most important being preservation of latencies from centrally- and peripherally-derived ABR waves. In addition, hearing thresholds are unchanged and wave amplitudes are not systematically suppressed, although they exhibit greater variability.ConclusionsWe demonstrate that 375 mg/kg 2,2,2-tribromoethanol followed after 5 min by 200 mg/kg chloral hydrate provides an anesthesia time of 60 min, has negligible effects on ABR wave latencies and thresholds and non-systematic effects on amplitudes.