Lamb JIVET Technology And The Expression Of Related Gene In The IVM

Juvenile in vitro embryo transfer(JIVET)is the integration of young female Supero vulation technique, oocytes matured in vitro techniques, invitro fertilization, embryo in vitro culture techniques and propagation technology of embryo transfer technology and aseries of biotechnology as a whole system. JIVET technology on young female ovaries of egg cells as a source of eggsin vitro embryo production to reduce the generation interval of livestock breeding, fully worthy of good genetic characteristics of femalere productive potential, improving the speed and productivity of livestock genetic improvement, provides a wealth of genetic resources for livestock breeding. In vitro maturation JIVET technology is an important part of, by regulating the hormone concentrations and other means to improve the effect of in vitro maturation in order to achieve cytoplasm and nucleus of mature common goals, significantly increased late embryonic growth and development potential, greatly increase the productivity of JIVET technology.This study in Jiangsu Province, the local white goat lamb as the main object of study, lamb slaughterhouse after oocyte in vitro maturation, and then on the development of mature oocyte in vitro fertilization. In the maturation of different concentrations of FSH and LH(2.5,5.0,7.5, 10.0,12.5μg/mL) to research on FSH and LH affect oocyte maturation rate, in order to improve the training system and compared late embryonic development of the Lambs and ewes, to explore the regulatory mechanism of oocyte maturation. This test using measuring sequence technology filter effect lamb eggs mother cell mature of key gene, and through real-time fluorescent quantitative technology on filter out of five a gene(MOS, RPS6KA1,CPEB1, ANAPC13 and CDK1)for quantitative validation, from molecular level research eggs mother cell mature of regulation mechanism, to improve cell pulp of mature rate then improve lamb eggs mother cell development to late embryos of potential. The results of this study are as follows:1. Concentration in the maturation of 2.5,5.0,7.5,10.0,12.5μg/mL FSH and LH, after maturation, the result shows maturation media supplemented with different concentrations of FSH and LH on oocyte maturation rate has a significant effect. Add 5.0μg maturation g/mL FSH and LH concentrations lamb oocyte maturation rate significantly higher than the other four groups; Add 10.0 μg maturation g/mL FSH Ewe, oocy-te maturation and LH concentration was significantly higher than the other four group and lambs and adult sheep oocytes cultured in the same maturation effects no significant difference (P>0.05).2. In same conditions nakedness outside fertilization Hou, on lamb and adult Ewe eggs mother cell of eggs crack rate, and 4 cell rate, and 8 cell rate, and 16 cell rate, and blastocyst rate for statistics antiwash, its results displayed lamb and adult Ewe eggs mother cell of eggs crack rate, and 4 cell rate no significantly differences (P>0.05), lamb eggs mother cell of 8 cell rate significantly below adult Ewe (P<0.05),lamb eggs mother cell of blastocyst rate very significantly below adult Ewe (P<0.01).3. By single cell sequencing technology on the lamb and Ewe, mature oocytes for transcriptome analysis, filtering out of MOS, RPS6KA1,CPEB1, ANAPC13 and CDK1 five important expression of genes, and by real-time fluorescence quantitative transcriptome sequencing data is verified for accuracy.