Effect of kdm4aa Gene on Low Temperature Tolerance of Zebrafish
In order to study the function of kdm4aa on low temperature tolerance of zebrafish,the levels of kdm4 families in kdm4aa-/-zebrafish and the level of kdm4aa in wild type(WT)zebrafish treated by low temperature were detected by real time quantitative PCR.The locomotor ability of kdm4aa-/-juvenile fish at low temperature was detected by the behavior trajectory tracking system of zebrafish.The low temperature tolerance of kdm4aa-/-zebrafish was measured by a stepwise cooling method.Reactive oxygen species(ROS),a-denosine triphosphate(ATP),gamma histone H2A.X(γH2A.X),and microtubule-associated protein 1 light chain 3(LC3)levels in kdm4aa-/-zebrafish were determined by DCFH-DA fluorescent probe,ATP detection kit and western blot,respectively.Real time quantitative PCR results showed that compared with WT zebrafish,the mRNA levels of kdm4ab and kdm4b in kdm4aa-/-zebrafish were significantly increased,while no significantly different of kdrn4c was observed.The detection of zebrafish behavior trajectory tracking sys-tem showed that the movement distance of juvenile zebrafish increased significantly after kdm4aa knockout at a low temperature of 18 ℃.At a low temperature of 8 ℃ mRNA level of kdm4aa in wild type zebrafish was significantly increased.Compared to WT zebrafish,the median lethal time LT50 of kdm4aa-/-zebrafish was significantly decreased after low temperature treatment(P＜0.000 1);ROS level was significantly increased(P＜0.0001);ATP content was significantly decreased(P＜0.01);The level of γH2A.X protein increased;LC3 protein levels decreased.These results showed that ROS accumulation of kdm4aa-/-led to decreased ATP synthesis and increased DNA damage,and the autophagy ability of kdm4aa-/-zebrafish was decreased,thus reducing the low temperature tolerance of zebrafish.
kdm4aa geneDanio rerioLow temperature toleranceDNA damage
NETL
NSTL
万方数据
维普
