Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica ›› 2015, Vol. 37 ›› Issue (5): 501-507.doi: 10.3881/j.issn.1000-503X.2015.05.003

• Orginal Article • Previous Articles     Next Articles

Regulatory Role of Nitric Oxide in Development and Hatching of Mouse Blastocysts

Xiao-yan PAN1, Zhi-xin LI1(), Xi-yan WANG1, Xue-nan WANG2, Jian-xin SUN1, Meng-tong ZANG1, Wen-jun LI1, Hong-he WANG1, Zhao-hua DOU1   

  1. 1Department of Histology and Embryology,Jilin Medical College,Jilin,Jilin 132013,China
    2Reproductive Medicine Center,the Affiliated Hospital of Jining Medical College,Jining,Shandong 272029,China
  • Received:2014-11-17 Online:2015-10-15 Published:2015-10-31
  • Supported by:
    Supported by the “Twelfth Five-Year” Scientific and Technological Research Project of Education Department of Jilin Province [吉教科合字(2013)第349号],the Natural Science Foundation of Shandong Province(ZR2011HL005),the Scientific and Technological Research Project of Jilin Province [201201077],the College Students’ Scientific Research Fund of Jilin Medical College [吉医学科字(2012)第04号],and the College Students’ Innovative and Entrepreneurial Training Program of Jilin Province[(2013)第850号]

Abstract:

Objective To determine the regulatory role and mechanism of nitric oxide (NO) in the development and hatching of mouse blastocysts. Methods The Kunming female mice were superovulated and then mated with mature male mice. On the day 2.5 of their pregnancy,morulae were flushed from their uterine horns with culture media. Morulae were cultured in different concentrations of N-nitro-L arginine methyl ester (L-NAME),sodium nitroprusside (SNP),or the combination of L-NAME and SNP in culture media for 48 hours. The development and hatching of blastocysts were examined on day 4 and day 5 and the total numbers of blastocyst cells and cysteinyl aspartate specific proteinase 3 (caspase 3) were observed under confocal laser scanning microscope. Results With the increase of the concentration of L-NAME or SNP,the hatching rate of blastocysts and the total number of blastocyst cells were significantly reduced. The addition of 10 nmol/L SNP in culture media with 5 mmol/L L-NAME significantly increased the development of blastocysts and promoted hatching of blastocysts. However,with increase of SNP concentration in culture media with 5 mmol/L L-NAME,the development and hatching rates of blastocysts were significantly decreased. L-NAME had no obvious effect on the expression of active caspase 3 in blastocyst cells. However,when being above 500 nmol/L,SNP significantly increased the expression of caspase 3 in blastocyst cells. Conclusions NO plays an important role in development and hatching of mouse blastocysts. Excessively high or low NO can damage the division of blastomeres,resulting in the failure of the blastocyst development and hatching. Also,excessively high NO can lead to the apoptosis of the blastocyst cells.

Key words: mouse, nitric oxide, blastocyst development, blastocyst hathing, apoptosis

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