Effects On The Spermatogenic Cells Of Eriocheir Sinensis By Atrazine Exposure

Eriocheir sinensis commonly known as crab, is an important economic aquaculture animal in China. In this study, E. sinensis was used as experimental material. The reproduction toxicity effects of atrazine on the spermatogenic cells were detected. Different concentration(0.001, 0.01, 0.1 and 1 mg/L) was setted in the experiment. Some methods were used, such as cell culture in vitro, flow cytometry(FCM), transmission electron microscopy,to investigate cell cytotoxicity, plasma membrane integrity, sperm acrosome reaction rate and the ultrastructure of sperm chromatin.The results are as follows:1. The LDH release data of crab spermatogenic cells dealt with herbicide indicated that,at all experimental concentrations, the content that LDH released to culture medium showed no significant difference(P> 0.05) compared to control group.2. The result of MTT assay showed that there was no significant difference(P> 0.05) in the viability of spermatogenic cells compared to control group after two hours atrazine exposing.3. Fluorescein diacetate- propidium iodide double staining and FCM were used in the investigation of plasma membrane integrity of crab sperm exposed to herbicide. The results showed that there was no markedly differential in statistics among all the groups(P> 0.05).4. Eosin staining was used in the research of acrosome reaction rate of E. sinensis sperm exposed to atrazine. The statistics data presented that there were no remarkable difference compared to control group after one hour atrazine expose(P> 0.05).5. Sperm chromatin dispersion(SCD) and agarose gel electrophoresis were used in the evaluation of the chromatin change of spermatophore exposed to herbicide.(1) The sperm chromatin in spermatophore of control group was uniform and orderly chromatin fiber, while the sperm chromatin of exposed group was damaged aggravatingly as the dose increased. The morphology of sperm chromatin changed obviously as the dose increased, such as the chromatin fiber distributed disorderly, sharply reduced. Meanwhile, there were more nodular structure of high electron-density and more regiment massive increased as the dose increased.It was proved that atrazine can observably damage the ultrastructure of sperm chromatin. The most significant effect happened when the concentration of ATR was 1 mg/L in which ratedamaged sperm reached 100%.(2) There was only one electrophoretic bands obtained in agarose gel electrophoresis after esposing to ATR.In conclusion, 0.001?0.01?0.1?1 mg/L atrazine had no obvious cytotoxicity effect on E.sinensis spermatogenic cells and had no significant effect on acrosome reaction rate and plasma membrane integrity. However, atrazine can observably change the ultrastructure of sperm chromatin. It indicated that atrazine has a certain extent reproductive toxicity on male E.sinensis.