The Expression,Localization And Function Of Nlrp9b Gene In The Early Mouse Embyro

Mouse oocytes and early embryo were studied by this experiment as the research object, the methods of immunohistochemistry, qRT-PCR, Western blot,RNA interference and immunofluorescence were proposed to study the expression,localization and function of Nlrp9b in the oogenesis and the early embryonic development,using a variety of bioinformatics software was analyzed the amino acids,and the sequence of amino acids by homology modeling predict the three-dimensional structure of proteins on translation.The results are as follows:(1) RT-PCR and western blot analysis showed that Nlrp9b transcription content only were detected in ovarian and small intestine in multiple organizations, the expression of NLRP9B protein was limited in different follicular oocytes and added with the increasing of age in mice.NLRP9B protein main located in the cytoplasm of preimplantation embryos with laser confocal microscopy.(2) Silencing in the oocyte Nlrp9b although was specificity but did not affect the rate of oocyte maturation the GV stage oocytes,at the same time,in order to study the lack of paternal genome,the development of embryos after parthenogenetic activation embryos still developed into normal,simultaneously Nlrp9b protein decreased significantly but the development of embryo was not blocked in the zygote,the blastocyst population was no obvious difference,this result shows that Nlrp9b has nothing to do with the maturation of oocyte and early embryonic development.(3) The evolutionary relationship between NIrp9b and Nlrp9a、Nlrp9 was far away using bioinformatics software analysis,and we know the total length of Nlrp9b is 1003 bp Nlrp9b the isoelectric charge of proteins is 7.82, the molecular weight is 114802.16, polar amino acid accounted for 41.3%,helix of secondary structure accounts for nearly half 49.05%,there are two nucleotides DAPIN and NACHT and seven leucine repeated structure,NLRP9B did not contain and positioning signal sequence.but including some transmembrane region.The smooth implementation of this project,can reveal NLRP9B physiological function and regulation of the molecular mechanisms of early embryonic development of mice,for early embryonic development mechanism in mice to elucidate theoretical basis.