Screening Of Molecular Markers And Function Exploration During Rare Minnow Neural Development Process

Developmental Neural Toxicology(DNT) refers to during the early stages of development, exposed by potential neurotoxic substances, the normal development of the nervous system is affect, such as proliferation, differentiation, migration and death of neurons, neurite outgrowth, synaptogenesis and myelination. This influence can occur in any period in the process of development, and have a lasting nerve dysfunction. So far, with the development of science and technology and the improvement of living standards, more and more chemicals appear constantly, it is important to assess the DNT of these chemicals.The poverty in knowledge of neurodevelopment of Chinese rare minnow(Gobiocypris rarus) limited its application as a native model fish in neurotoxicital studies and risk assessment. Using rare minnow as the object of study, applying RT-PCR, we clone partial sequence of ngf, bdnf, gfap, mag and α1-tubulin, study the developmental expression map when embryos/larvae developing to the cleavage stage(0.5h), morula stage(3.5h), blastula stage(5.5h), gastrula stage(10h), neurula stage(13.5h), appearance of myomere stage(14.5h), appearance of optic rudiment stage(16h), formation of eye crystal stage(24h), heart beat stage(30.5h), hatching(61h), 4 dpf(day post fertilization), 5dpf, 6dpf, 7dpf, 10 dpf, 14 dpf and tissue distribution map of brain, spinal cord, eye, liver, gonad, muscle. The transcriptional profile revealed expression of ngf, bdnf, gfap, mag and α1-tubulin presented different trends from cleavage stage to 14 dpf, related to their functions during the process of neural development; highest abundance of ngf, bdnf, mag and α1-tubulin m RNA in the brain and of gfap in the spinal cord. Exposed with organophosphorus pesticide CPF and organophosphorus ester TPP, these five genes in embryos are more susceptible than adults, the toxic effect of CPF is higher than TPP when exposing to adults. Gfap and mag maybe become potential biomarkers. These results provided a foundation for future studies in neurodevelopment and neurotoxicity assessment and mechanism research using rare minnow as a native model fish.