Effects of Crh on Redox Homeostasis of Zebrafish Danio rerio Embryos
WU Chunan, LYU Weiqun, CHEN Aqin
1. National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China; 2. Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Ministry of Education, Shanghai Ocean University, Shanghai 201306, China
Abstract:In order to investigate the effect of corticotropin-releasing hormone (Crh) on redox homeostasis in fish embryos, 1-day-old embryos of AB wild-type zebrafish Danio rerio were treated with 0.05% DMSO (control, CTL), 20 nmol/L Crhα and 20 nmol/L Crhβ at (28±1) ℃. The changes in reactive oxygen species and glutathione (GSH) contents were measured in zebrafish embryos, and the changes in the expression of Crh receptors (crhr1, crhr2) and antioxidant-related genes were detected by real-time quantitative PCR (qRT-PCR) at 2, 3, 4 and 5 days of age, respectively. The results showed that there was significant decrease in hatching rate of zebrafish embryos exposed to both Crhα and Crhβ at 54—78 hpf (hour post-fertilization) (P<0.05), with significant increase in reactive oxygen species (ROS) content in the zebrafish embryos exposed to Crhα at 3—5 dpf (days post-fertilization). The qRT-PCR analysis results revealed that Crhα and Crhβ significantly inhibited the GSH synthesis and the ggt1b expression(P<0.05), and the crhr1 mRNA level showed a trend of first decreasing and then increasing with embryonic development. The findings provide new insights into the regulatory role of Crh in redox homeostasis and have certain reference significance for exploring the physiological responses of teleost fish to stress.
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