| The zebrafish, Danio rerio, is a small tropical teleost, which sensitive to aconsiderable part of chemical pollutant. It was recommended as standard fish modelof toxicology tests by ISD and State Bureau of Environmental Protection. Thezebrafish genome project showed that genes of human and zebrafish is highlyhomologous, which made zebrafish, as a new vertebrate model of human disease, isgaining increasing interest. Parkinson’s disease （PD）, the world’s second largestneurodegenerative diseases, the incidence rate in people over65years old up to1%.However, the pathogenic mechanism of PD is still unknown and still can notcompletely cure, so the study on animal PD model have contributed to pathogenicmechanism and treatment strategies of PD.This study used embryos, larvae and adult zebrafish as the research object,established zebrafish PD model by haloperidol （Hal）, explored the neuroprotectiveeffect of Antarctic krill oil on adult zebrafish PD model. Firstly, the neurotoxicity ofHal on zebrafish embryos and larvae are studied, feasibility of Hal induced zebrafishPD model is determined primarily; then acute toxicity and histopathology research ofadult zebrafish were carried on, aimed at comparative advantages of larvae and adultzebrafish, at the same time decided the dose of Hal; Lastly, the neuroprotectiveeffect of Antarctic krill oil on adult zebrafish PD model was studied.Fertilized eggs of zebrafish were collected, experiment was performed in6-wellplates.0.01%DMSO was used to increase the solubility of Hal, setted normal controlgroup, DMSO group, Hal0.10,0.20,0.40,0.80and1.60mg·L-1. Embryos were putinto illuminating incubator, regular observe the morphological development of eachgroup embryos, counted the rates of incubation, aberration and death of zebrafishembryos. Immunostaining was used to observe the changes of DA neurons in4dpflarvae zebrafish, then swimming speed of7dpf larvae were determined. Researchrevealed that Hal made zebrafish embryos developmental delay, and high dose Hal lead to aberration and death; locomotor activity of larvae became lower as theconcentration of Hal increase, and the larvae zebrafish showed rigidity and incline,which were similar to PD, while these symptoms were improved by L-dopa; Halcould induced DA neurons loss in larvae zebrafish. The results indicated that Halcould be used to establish zebrafish PD model.Concentrations of Hal for acute toxicity experiment of adult zebrafish,0.243,0.300,0.371,0.458,0.566and0.700mg·L-1, were determined by prepare experiments.48h and96h LC50of Hal were0.388and0.325mg·L-1. Besides, zebrafish showedPD symptoms, such as rigidity and incline. HE stain of brain, liver and kidney wereperformed, results showed that brain tissue of zebrafish treated with Hal weredamaged more serious than liver and kidney. Because Larvae body is too small in size,ethological changes are difficult to identify, and it is uncertain whether the loss of DAneurons is induced by inhibition of neuron development or damaged of matureneurons, so adult zebrafish is more suitable for PD model by contrast to larvae.Hal0.3mg·L-1induced adult zebrafish PD model, locomotor activity wereassessed by counting the number of lines for5min that adult zebrafish crossed,opercular ventilation and pigmentation were counted also. Results showedthat locomotor activity of Hal treated zebrafish decreased and performed typical PDsymptoms. Moreover, DA neurons reduced of Hal0.3mg·L-1zebrafish. Experimentsdemonstrated that adult zebrafish is ideal for establishment of PD model.Zebrafish were transferred into Antarctic krill oil100mg·L-1after treated withHal, locomotor activity of zebrafish was significantly improved, opercular ventilationand pigmentation also tended to be normal. Immunostaining showed DA neuronsobviously increase compared with PD zebrafish. Preliminary showed that damagedDA neurons could be repaired by Antarctic krill oil to a certain extent. In other words,Antarctic krill oil has neuroprotective and neuroreparable effects for zebrafish. |
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