ABSTRACT
Objective: To investigate the effect of asarone on the zebrafish embryonic development and behavior. Methods: The 3 hours post fertilization (hpf) zebrafish embryos were exposed to different concentrations (0, 25,50,100, 200 and 400 μmol/L) of asarone solution. The asarone solution was replaced each 24 h. Microscope was used to observe embryos morphology at 24,48,72 and 96 hpf, respectively. Spontaneous movements at 24 hpf. heart rate at 48 hpf, hatch rate, deformity rate and mortality rate were evaluated.The sepnl gene expression of zebrafish in 96 hpf was detected by RT-qPCR. With Noldus tracking system the behaviors of zebrafish larvae exposed to asarone were recorded. Results: After zebrafish embryos were exposed to the different concentrations of asarone. their spontaneous movements were decreased. Compared with the control group, the 100,200 and 400 μmol/L groups were significantly decreased (P<0.01). Tłiere was a significant difference (P<0.01) in 48 hpf heart rate between all asarone groups and control group. Spine curvature, pericardial edema, yolk sac edema and other deformity were observed at 72 hpf and 96 hpf. Compared with the control group, the 100, 200 and 400 μmol/L groups showed significant decreased hatch rate (P<0.01). There was a significant difference (P<0.05) in mortality rate between the 400 μmol/L group and control group. With concentrations of asarone increasing, the speed and distance of movement of 200 and 400 μmol/L group were significantly reduced compared to the control group (P<0.05). There was a significant difference between the 200.400 μmol/L groups and the control group in activity (P< 0.01).At 96 hpf. the expression of sepnl gene in arone groups were decreased compared to control group, especially the 100 μmol/L group (P<0.05), and the 200 and 400 μmol/L groups (P<0.01). Conclusion: Asarone had teratogenic effect on zebrafish embryos and larvae, and inhibitory effect on larvae behavior .We should be careful to use asarone in infant medication.
ABSTRACT
Objective To investigate the effect of asarone on the zebrafish embryonic development and behavior. Methods The 3 hours post fertilization(hpf) zebrafish embryos were exposed to different concentrations(0, 25,50,100,200 and 400 μmol/L) of asarone solution. The asarone solution was replaced each 24 h. Microscope was used to observe embryos morphology at 24,48,72 and 96 hpf, respectively. Spontaneous movements at 24 hpf, heart rate at 48 hpf, hatch rate, deformity rate and mortality rate were evaluated.The sepn1 gene expression of zebrafish in 96 hpf was detected by RT-qPCR. With Noldus tracking system the behaviors of zebrafish larvae exposed to asarone were recorded. Results After zebrafish embryos were exposed to the different concentrations of asarone, their spontaneous movements were decreased. Compared with the control group, the 100,200 and 400 μmol/L groups were significantly decreased (P<0.01). There was a significant difference(P<0.01) in 48 hpf heart rate between all asarone groups and control group. Spine curvature, pericardial edema, yolk sac edema and other deformity were observed at 72 hpf and 96 hpf. Compared with the control group, the 100, 200 and 400 μmol/L groups showed significanly decreased hatch rate(P<0.01). There was a significant difference (P<0.05) in mortality rate between the 400 μmol/L group and control group. With concentrations of asarone increasing, the speed and distance of movement of 200 and 400 μmol/L group were significantly reduced compared to the control group(P<0.05). There was a significant difference between the 200,400 μmol/L groups and the control group in activity(P< 0.01).At 96 hpf, the expression of sepn1 gene in arone groups were decreased compared to control group, especially the 100 μmol/L group(P<0.05), and the 200 and 400 μmol/L groups(P<0.01). Conclusion Asarone had teratogenic effect on zebrafish embryos and larvae ,and inhibitory effect on larvae behavior .We should be careful to use asarone in infant medication.
ABSTRACT
OBJECTIVE To investigate the develop mental toxicity of muscone to embryos. METHODS With zebrafish embryos as a model,The 3 h post fertilization (hpf)embryos were exposed to muscone at 5,10,20,40,80 and 160 μmol·L -1 culture solutions for 96 h and inspected daily with mi-croscopy for larval morphology.The drug solution was replaced every 24 h.Spontaneous move ments were checked at 24 hpf.Heart rate at 48 hpf,hatching rate,e mbryo deformity rate and mortality rate were evaluated.The expression of sepn1 was determined with real-ti me quantitative PCR technique at 96 hpf.RESULTS The 24 hpf spontaneous move ments showed no significant difference.At 48 hpf, spine curvature,pericardial ede ma,yolk sac ede ma,and abnormal swi mming were observed.In addition, the 48 hpf heart beats(10 s)was decreased fro m 26.5 ±1 .0 to 18.0 ±1 .9(P <0.01 ).At 48 hpf , hatching rate of 5 ~40 μmol·L -1 decreased(P <0.05),while of 160 μmol·L -1 increased (P <0.05) co mpared with muscone 0 μmol·L -1 .Muscone had little effect on hatching rate at other ti me points;Mal-formation rate and mortality rate at higher concentrations were up to 100%.The sepn1 gene expression at 96 hpf in the exposure groups decreased co mpared with that of control group(P <0.01 ).CONCLU-SION Muscone had toxic effects on the develop ment of zebrafish embryos,including spine curvature, abnormal swi mming,and pericardial ede ma.These effects may be related to the inhibition of sepn1 gene expression by muscone.