돼지 초기배 발달에 있어서 ATF5 발현과 미토콘드리아 분열과의 상관관계 분석에 대한 연구

• 영문명: Analysis of the Correlation between ATF5 Expression and Mitochondrial Fission during Early Embryonic Development in Pigs
• 발행기관: 한국동물보건학회
• 저자명: 이민형(Min-Hyung Lee) 구덕본(Deog-Bon Koo)
• 간행물 정보: 『한국동물보건학회지』4권 2호, 139~159쪽, 전체 21쪽
• 주제분류: 의약학 > 기타의약학
• 파일형태: PDF
• 발행일자: 2025.06.30

국문 초록

Activating transcription factor 5 (ATF5) is a key factor of the mitochondrial unfolded protein response and plays a crucial role in maintaining mitochondrial function. Mitochondrial fission is one of the essential functions of mitochondria and plays an important role in cell survival. However, the relationship between ATF5 expression and mitochondrial fission during early porcine embryonic development remains unclear. In this study, mitochondrial-specific superoxide production and mitochondrial length during early embryonic development in pigs were analyzed by using Mito-SOX and MitoTracker Orange staining. As a result, mitochondrial reactive oxygen stress (ROS, p < 0.05) and fragmented mitochondria (p < 0.01) decreased during the cleavage stage. Additionally, the expression pattern of ATF5 was examined at different embryonic stages, and its co-localization with the nucleus and mitochondria was confirmed. Furthermore, to investigate the effect of mitochondrial fission inhibition, porcine zygotes were treated with the mitochondrial fission inhibitor 1 (Mdivi-1, 10 and 50 μM) for 2 days after in vitro fertilization. As expected, blastocyst developmental rate and expanded blastocyst formation were decreased in the Mdivi-1 treated porcine embryos. Treatment of embryos with Mdivi-1 led to a significant increase in DNA fragmentation and mitochondrial-specific superoxide production in blastocysts, as confirmed by the TUNEL assay (p < 0.01) and Mito-SOX staining (p < 0.001). Mitochondrial morphology analysis revealed a decrease in the number of fragmented mitochondria (<1 μm) and an increase in the number of elongated mitochondria (>3 μm) in the 10 μM and 50 μM Mdivi-1 treatment groups. Additionally, co-localization of ATF5 with the nucleus and mitochondria indicated that Mdivi-1 treatment enhanced ATF5 expression. Nuclear co-localization of ATF5 increased (p < 0.01), whereas its co-localization with mitochondria downregulated (p < 0.01). To further investigate the effect of mitochondrial fission, we analyzed the protein levels of mitochondrial fission-related proteins (DRP1, p-DRP1Ser616) and ATF5. As a result, the expression of p-DRP1Ser616 decreased in a dose-dependent manner, while ATF5 expression exhibited an increasing trend (p < 0.05). Finally, to examine the effect of ATF5 inhibition on blastocyst development and mitochondrial fission, we designed porcine-specific ATF5 siRNA and transfected it into porcine zygotes. In the ATF5 siRNA-treated group, mitochondrial superoxide generation (p < 0.001) and the number of elongated mitochondria increased (p < 0.05), while the co-localization of ATF5 with the nucleus and mitochondria decreased (p < 0.01). In conclusion, our findings suggest that ATF5 regulates mitochondrial fission, thereby maintaining mitochondrial function and influencing blastocyst development.

목차

1. 서론

2. 재료 및 방법

3. 결과

4. 고찰

References

키워드

ATF5 Mitochondrial fission DRP1 Blastocyst development Pig