OBJECTIVE: This study is to determine the regulatory role of nitric oxide in mouse blastocyst hatching. METHODS: Kunming female mice were superovulated and then mated with mature male mice. On day 2.5 of their pregnancy, the pregnant mice were killed and morulae were flushed from their uterine horns with culture media. Morulae were cultured in media with different concentrations of N-nitro-L arginine methyl ester (L-NAME), sodium nitroprusside (SNP), 8-Br-3'-5'-cyclic guanosine monophosphate (8-Br-cGMP) or the combination of L-NAME with SNP or 8-Br-cGMP for 48 h. The hatched blastocysts were examined on day 5 and the expressions of epithelial nitric oxide synthase (eNOS) and active cysteinyl aspartate specific proteinase 3 (caspase 3) were observed under confocal laser scanning microscope. RESULTS: L-NAME significantly reduced the expression of eNOS in blastocyst cells. With the increase of the concentrations of L-NAME, SNP or 8-Br-cGMP, blastocyst hatching rate was significantly lowered. In addition, 5 mM L-NAME, 2 μM SNP and 2 μM 8-Br-cGMP completely inhibited blastocyst hatching. Low concentrations of SNP or 8-Br-cGMP in culture media containing 5 mM L-NAME significantly reversed the inhibition of blastocyst hatching and promoted hatching development. Moreover, 5 mM L-NAME and 2 μM 8-Br-cGMP had no significant influence on the expression of active caspase 3 in blastocyst cells. SNP (> 500 nM) significantly increased the expression of active caspase 3 in blastocyst cells. CONCLUSIONS: NO/cGMP pathway plays an important role in mouse blastocyst hatching. Excessive or depleted NO can interrupt blastocyst hatching. Excessive NO leads to apoptosis of blastocyst cells.
OBJECTIVE: This study is to determine the regulatory role of nitric oxide in mouse blastocyst hatching. METHODS: Kunming female mice were superovulated and then mated with mature male mice. On day 2.5 of their pregnancy, the pregnant mice were killed and morulae were flushed from their uterine horns with culture media. Morulae were cultured in media with different concentrations of N-nitro-L arginine methyl ester (L-NAME), sodium nitroprusside (SNP), 8-Br-3'-5'-cyclic guanosine monophosphate (8-Br-cGMP) or the combination of L-NAME with SNP or 8-Br-cGMP for 48 h. The hatched blastocysts were examined on day 5 and the expressions of epithelial nitric oxide synthase (eNOS) and active cysteinyl aspartate specific proteinase 3 (caspase 3) were observed under confocal laser scanning microscope. RESULTS:L-NAME significantly reduced the expression of eNOS in blastocyst cells. With the increase of the concentrations of L-NAME, SNP or 8-Br-cGMP, blastocyst hatching rate was significantly lowered. In addition, 5 mM L-NAME, 2 μM SNP and 2 μM 8-Br-cGMP completely inhibited blastocyst hatching. Low concentrations of SNP or 8-Br-cGMP in culture media containing 5 mM L-NAME significantly reversed the inhibition of blastocyst hatching and promoted hatching development. Moreover, 5 mM L-NAME and 2 μM 8-Br-cGMP had no significant influence on the expression of active caspase 3 in blastocyst cells. SNP (> 500 nM) significantly increased the expression of active caspase 3 in blastocyst cells. CONCLUSIONS: NO/cGMP pathway plays an important role in mouse blastocyst hatching. Excessive or depleted NO can interrupt blastocyst hatching. Excessive NO leads to apoptosis of blastocyst cells.
Authors: Priscila Di Paula Bessa Santana; Thiago Velasco Guimarães Silva; Nathália Nogueira da Costa; Bruno Barauna da Silva; Timothy Frederick Carter; Marcela da Silva Cordeiro; Bruno José Martins da Silva; Simone do Socorro Damasceno Santos; Anderson Manoel Herculano; Paulo Roberto Adona; Otávio Mitio Ohashi; Moysés dos Santos Miranda Journal: Mol Reprod Dev Date: 2014-09-18 Impact factor: 2.609
Authors: Laura C Kusinski; Joanna L Stanley; Mark R Dilworth; Cassandra J Hirt; Irene J Andersson; Lewis J Renshall; Bernadette C Baker; Philip N Baker; Colin P Sibley; Mark Wareing; Jocelyn D Glazier Journal: Am J Physiol Regul Integr Comp Physiol Date: 2012-05-02 Impact factor: 3.619
Authors: Denise R S Fabri; Sara C Hott; Daniel G Reis; Caroline Biojone; Fernando M A Corrêa; Leonardo B M Resstel Journal: Eur Neuropsychopharmacol Date: 2014-08-11 Impact factor: 4.600
Authors: Renan O Silva; Larisse T Lucetti; Deysi V T Wong; Karoline S Aragão; Eudmar M A Junior; Pedro M G Soares; André Luiz R Barbosa; Ronaldo A Ribeiro; Marcellus H L P Souza; Jand-Venes R Medeiros Journal: Nitric Oxide Date: 2014-05-12 Impact factor: 4.427