بيان كمي ژن هاي BAX,BAG1 و BCL-2 در جنين هاي انساني با درجات مختلف فراگمنتاسيون حاصل از ART

Quantitative Expression of BAG1, BAX and BCL-2 genes in Human Embryos with Different Fragmentation Grades Derived from ART

Objective: The purpose of this study was to evaluate the quantitative expressions of BAG1, BAX and BCL-2 in human embryos with different fragmentation grades as derived from assisted reproduction technology (ART). Materials and Methods: Fragmented and normal human 8-cell embryos were scored according to the degree of fragmentation with an inverted microscope and divided into four grades (grade I: no or minimal fragmentation (<5%), grade II: embryos with <25% fragmentation, grade III: embryos with >25% fragmentation and grade IV: apoptotic induced embryos with actinomycin D). In this study, TUNEL labeling was initially used to detect apoptosis, and then revers transcription polymerase chain reaction (RT-PCR) and quantitative PCR were used to define the quantitative expressions of experimental genes in human embryos with different fragmentation grades. Results: The results of TUNEL labeling showed that embryos with higher fragmentation had a high number of apoptotic bodies. The results of RT-PCR and q-PCR analyses showed a significantly decreased amount of BAGI transcript expression from group I to group IV. The highest expression of BAX gene was observed in group II, however, the transcript of BCL-2 gene was not observed in any of the experimental groups. The effect of actinomycin D on transcript expression amounts of experimental genes in apoptotic induced embryos (group IV) compared to control embryos (group I) showed a significant decrease. Conclusion: mRNA expression of BAG1 gene can be used as a good marker to detect apoptosis in human embryos. However, the transcript of BCL-2 gene does not play a role in the detection of apoptosis in human embryos at the 8-cell stage.