Abstract:In order to study the sequence characteristics of heat shock 70 kD protein-12B gene (HSPA12B) and its expression in yak (Bos grunniens) early in vitro fertilization (IVF) embryos, the specific primers were designed by the conservative sequence of HSPA12B gene of the cattle (Bos taurus),and the gene sequence could be obtained by cloning. The HSPA12B sequence was analyzed by bioinformatics. The HSPA12B in early IVF embryos were detected by the methods of quantitative Real-time PCR (qRT-PCR). The results of bioinformatics showed the coding region of the yak HSPA12B gene was 705 bp in length, encoding 234 amino acids, and the content of Alanine resduces was the highest (about 10.3%) and the content of Asparagine resduces was the lowest (about 0.9%). According to homology analysis, the similarity of HSPA12B gene in yak and cattle was up to 99.5%, showing the HSPA12B gene was highly conserved. The protein was hydrophilic and had no signal peptide, which was not a transmembrane and secreted protein. qRT-PCR results showed that the HSPA12B in yak early embryos fertilized in vitro expressed in each period, and the relative expression of HSPA12B in 2-4 cell stage embryos was 14.814 8, 4.269 9 and 40.485 8 folds respectively, compared to the 5-8 cell stage embryos, morulae and blastocysts. The difference of the relative expression of HSPA12B between 2-4 cell stage embryos and 5-8 cell stage embryos,morula and blastocysts was very significant (P<0.01). These results showed HSPA12B played an important role in the development of embryos IVF and can inhibit cell apoptosis, cell senescence, and regulate embryonic development.
Obrenovitch T P. Molecular physiology of preconditioning-induced brain tolerance to ischemia. Physiological Reviews, 2008, 88:211-247.Meimaridou E, Gooljar S B, Chapple J P. From hatching todispatching: the multiple cellular roles of the Hsp70 molecular chaperone machinery. Journal of Molecular Endocrinology, 2009, 42:1-9.Su C Y, Chong K Y, Owen O E, Dillmann W H, Chang C, Lai C C. Constitutive and inducible hsp70s are involved in oxidative resistance evoked by heat shock or ethanol. Journal of Molecular and Cellular Cardiology, 1998, 30: 587-598.Fei G, Guo C, Sun H S, Feng Z P.HSP70 reduces chronic hypoxia-induced neural suppression via regulating expression of syntaxin. Advances in Experimental Medicine and Biology, 2008, 605:35-40.Ettore Beghi.Overview of Studie to Prevent posttraumatic Epilepsy[J].Epilepsia,2003,44(suppl.10):21-26.Zhihua Han, Quynh A. Truong, Shirley Park.Two Hsp70 family members expressed in atherosclerotic lesions.Medical sciences,2003,100:1256-1261.Binder RJ.Heat shock protein Vaccins:from bench to beside[J].Int RevImmunol,2006,25(5-6):353-357.Hu G,Tang J,Zhang B,Lin Y,Hanai J,Galloway J,Bedell V,Bahary N,Han Z,RamchandranR,Thisse B,Thisse C,Zon LI,Sukhatme VP(2006)A novel endothelial-specific heat shock protein HSPA12B is required in both zebrafish development and endothelial functions in vitro.J Cell Sci 119:4117-4126.Steagall RJ,Rusinol AE,Truong QA,Han Z(2006)HSPA12B is predominantly expressed in endothelial cells and required for angiogenesis.Arterioscler Thromb Vasc Biol 26::212-2018.李竞进. HSPA12B对慢性心梗后心功能不全和心肌重塑的保护作用[D].南京医科大学,2011.(Li J J. Protective effect of HSPA12B on cardiac dysfunction and myocardial remodeling in chronic myocardial infarction[D]. Nanjing Medical University,2011) 朱勇飞.三个Hsps家族在小鼠器官发育中的表达特点和在肢、腭异常发育中的作用[D].第二军医大学.(expression characteristic of three Hsps gene families in the organs of mice during the abnormal development of palate and limb[D]. The Second Military Medical University)余桂芳.HSPA12B对内毒素刺激的小鼠肺微血管内皮细胞的作用及其机制[D].第二军医大学,2013.(The mechanism of HSPA12B on microvascular endothelial cells of endotoxin stimulated mouse pulmonary [D]. The Second Military Medical University)Liping You, Xuanxuan Ning, Feng LiuThe.response profi les of HSPA12A and TCTP from Mytilus galloprovincialis to pathogen and cadmium challenge. Fish & Shellfish Immunology,2013,35:343-350.Lihua Kang,Guowei Zhang ,Yaohua Yan.The Role of HSPA12B in Regulating Neuronal Apoptosis.Neurochem Res ,2013, 38:311–320.唐桂娥. 体内和体外不同发育阶段小鼠胚胎内部活性氧(ROS)的测定[D].延边大学,2006.(Tang G E.The detection of ROS in different developmental stages of mouse embryo in vivo and in vitro[D]. Yan Bian University,2006)张全乐. 胚胎碎片及发育阻滞与活性氧相关性研究[D].郑州大学,2012.(Zhang Q L.Studies the relationship between active oxygen and Embryo fragmentation、development block[D].Zheng Zhou University,2012.)