Effect of Maternal Diabetic Environment on Development of Early Mouse Embryo by in vitro Fertilization

doi:10.3969/j.issn.1674-5817.2015.02.014

Abstract

Abstract: Objective The experimental mice were induced to be diabetes mellitus (DM) mice through streptozotocin (STZ) intraperitoneal injection, in order to study the effect of maternal hyperglycemia on early mouse embryo development by in vitro fertilization. Methods ICR mice aged 6~8 weeks were intraperitoneally injected with STZ to established DM models, and the normal mice were considered as control group . The model mice were superovulated by intraperitoneal injection with PMSG and hCG, then the oocytes were fertilized with sperms from normal male mice in vitro. The rates of fertilization were counted and 2-cell embryos were transferred to pseudopregnant mice. Finally 14- dold embryos were obtained to extract. total RNA. After detected by Agilent 2100 Bioanalyzer system, total RNA were reversed transcription, labeled, hybridized, eluted and the data of microarray was scanned by Affymetrix Scanner 3000. Basic data were read by Command Console Software 3.1. The experiments were repeated three times, then the expression of genes between treated group and control group were compared. Results The ratio of two-cell embryo in hyperglycemia group (52.3%), was significantly lower than that in the control group (77.2%) (P<0.01). A total of 121 differentially expressed genes (P<0.05) were screened out. One hundred and nineteen genes expression levels of the experimental group was less 0.5 times than those of the control group, while 2 genes showed a more than 2-fold up regulation. Conclusion After oocytes of model mice fertilizing with sperms of normal male mice in HTF, the rate of embryo developing to two-cell was lower than normal mice. The higher of blood glucose level, the lower of two-cell rate. The growth and development of the oocytes were affected by maternal diabetes environment through up-regulating the metabolism-related genes and down-regulated development-relating genes .

Key words: Streptozotocin(STZ), In vitro fertilization, Microarray, Differentially expression

CLC Number:

Q95-33

QIAANG Su jing, TAO Ling-yun, LIU Li-jun, XU Ping, GAO Cheng. Effect of Maternal Diabetic Environment on Development of Early Mouse Embryo by in vitro Fertilization[J]. Laboratory Animal and Comparative Medicine, 2015, 35(2): 155-160.

References

[1] 邵伟娟, 陶凌云, 赵茹茜, 等. 不同剂量STZ 诱导小鼠糖尿病模型及生殖能力的研究[J]. 上海交通大学学报: 农业科学版, 2007, 25(6):541-546.
[2] Sun FY, Kawasakia E, Akazawac S.Apoptosis and its pathway in early post-implantation embryos of diabetic rats Diabetes[J]. diabetes Res Clin Pract, 2005, 67(2):110-118.
[3] Vesela J, Cikos S, Hlinka D, et al.Effects of impaired insulin secretion on the fertilization of mouse oocytes[J]. Hum Reprod, 1995, 10(12):3233-3236.
[4] 林锟. 高血糖对卵母细胞和早期胚胎细胞凋亡的影响[J]. 医学综述, 2009, 15(19):2932-2934.
[5] Okamura K, Wintle RF, Scherer SW.Characterization of the differentially methylated region of the Impact gene that exhibits Glires-specific imprinting[J]. Genome Biol, 2008, 9(11):160-170.
[6] 刘齐, 王燕, 陈岩, 等. Grb10 基因在小鼠胚期特异性表达分析[J]. 遗传, 2009, 31(7):732-740.
[7] 胥传飞, 卢晟盛, 王友亮, 等. 接头蛋白Gabl的结构及功能研究进展[J]. 生物技术通讯, 2009, 20(6):839-842.
[8] Shiura H, Nakamura K, Hikichi T, et al.Paternal deletion of Meg1/Grb10 DMR causes maternalizationof the Meg1/Grb10 cluster in mouse proximal chromosome11 leading to severe pre- and postnatal growth retardation[J]. Hum Mol Genet, 2009, 18(8):1424-1438.
[9] Park CH, Uh KJ, Mulligan BP, et al.Analysis of imprinted gene expression in normal fertilized and uniparental preimplantation porcine embryos[J]. PLoS One, 2011, 6(7):e22216.
[10] Frank D, Fortino W, Clark L, et al.Placental over growth in mice lacking the imprinted gene IPI[J]. Proc Natl Acad Sci USA, 2002, 99(11):7490-7495.
[11] Himes KP, Koppes E, Chaillet JR.Generalized disruption of inherited genomic imprints leads to wide-ranging placental defects and dysregulated fetal growth[J]. Dev Biol, 2013, 373(1):72-82.