Circadian Rhythm of Type-C Natriuretic Peptide Concentration in Rabbit Aqueous Humor

A. Paranhos-Jr, O. Koji, P.A Mello and D. Gregory

Federal University of Sao Paulo, Brazil., Yale University, CT-USA

Objective: to evaluate the circadian rhythm of type-C Natriuretic Peptide (CNP) concentration in rabbit aqueous humor. Methods: 41 male New Zealand white rabbits were entrained to a lighting schedule of alternating 12 h period of light and dark for at least 2 weeks prior to the beginning of the experiment. Lights on was defined as 00:00 circadian time (CT) and lights off at 12:00 CT. Animals were killed at 6 intervals (Light: 2h, 6h, 10h, Dark: 14h,18h, 22h), four animals for each interval except for interval 6h (11 animals) and 22h (12 animals). Aqueous was collected, anti-proteases (1 ml EDTA/2mM, 1 ml PMSF/2 mM and 5ml pepstatin 0,2 mM) were added and the samples stored at-84ºC.CNP was measured by CNP-22 radioimmunoassay. Statistical analyses: ANOVA, unpaired t-test and linear regression. Results: The CNP concentrations in pg/200ml were not statistically different when rabbits were divided only into light (mean: 2.62 ±0.91) and dark groups (mean: 3.02 ±1.16). Statistical differences were also not found when the 6 intervals were compared (F= 1.72 P=0.157), where the highest differences on concentration were found between 22h (3.41 ±1.24) and 10h (mean:2.20 ±0.72). There was no significant tendency of change concentration over time (r2=7.4%, p=0.085). Conclusion: The CNP concentration in rabbit aqueous humor does not have a significant fluctuation over circadian time.