Abstract
Whole body plethysmography (WBP) is used to quantify pulmonary function in conscious, unrestrained mice. We determined currently whether time of day and environmental lighting influence day-to-day reproducibility of pulmonary function, and quantifed the necessary habituation time in the WBP chamber. Two-month-old male C57BL6 and mdx mice (n = 8/group, reverse light cycle), were examined on consecutive days using a calibrated WBP chamber and manufacturer software was used to calculate respiratory measures. Respiratory data stabilized between 5-10 min for all variables. Mice exhibited time of day respiratory differences, performing more forceful and less frequent breaths midday (11:45 a.m. and 3:00 p.m.) compared to 7:30 a.m. WBP performed in darkened conditions elicited more forceful breathing than lit conditions. Day-to-day reproducibility during controlled conditions ranged from r(2) = 0.58 to 0.62 for the functional measures. Findings indicate reproducible respiratory data are obtainable following a 15-min chamber habituation and standardization of time of day and room lighting.
Highlights
Whole body plethysmography (WBP) has been used for decades to estimate respiratory function in animals [1,2,3]
Interest in WBP has proved important for mouse models where animal size and temperament are not conducive to direct determination of pulmonary function [4,5,6]
No strain-dependent differences were noted between C57 and mdx mice, and as such, inter-strain data were combined for these calculations
Summary
Whole body plethysmography (WBP) has been used for decades to estimate respiratory function in animals [1,2,3]. Interest in WBP has proved important for mouse models where animal size and temperament are not conducive to direct determination of pulmonary function [4,5,6]. While determination of respiratory rate is not overly problematic, accurate tidal volume (VT) estimates are influenced dramatically by subtle changes in pulmonary mechanics and fluctuations in barometric pressure (reviewed in [7]). The advantage of WBP over invasive respiratory measures is that WBP outcomes are not confounded by animal restraint or anesthesia [7]. WBP has been used extensively to gain serial measures of respiratory function in a variety of mammalian models of disease, including mouse strains [4, 10,11,12,13,14]
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