Earticle

초록

영어

Background: The 75-kDa glucose-regulated protein (GRP75) plays a crucial role in regulating the formation of mitochondria-associated endoplasmic reticulum (ER) membranes (MAMs), facilitating the transfer of Ca2+ ions, and is essential for lipid metabolism, mitochondrial function, calcium homeostasis, and apoptosis in mammalian cells. However, the relationship between GRP75 expression and preimplantation embryonic development in pigs remains unknown. Methods: In this study, we investigated whether GRP75 influences ER–mitochondrial junctions and mitochondrial Ca2+ levels in porcine embryos in vitro . We examined the expression of GRP75 at the zygote, cleavage, and blastocyst stages using immunofluorescence staining and western blot analysis. Results: GRP75 fluorescence and mitochondrial Ca2+ levels were significantly lower (p < 0.01) in the blastocysts than in the zygotes. Western blot analysis revealed a decline in the expression of mitochondrial fusion factors mitofusin 2, GRP75, and the mitochondrial calcium uniporter complex MICU1 protein at the blastocyst stage. To investigate the effects of GRP75 on blastocyst developmental competence, porcinespecific GRP75-siRNA (25 nM) was microinjected at the zygote stage. The results showed a significant decrease in the development capacity until the blastocyst stage (Control: 31.2 ± 2.0%, N.C. siRNA (25 nM): 29.8 ± 3.1%, vs. GRP75-siRNA (25 nM): 24.1 ± 1.6%; p < 0.05). GRP75 in the mitochondria and ER-localized GRP75 were both significantly reduced in blastocysts of pigs microinjected with GRP75 siRNA. Along with ER–mitochondrial colocalization, the MAM formation ratio was significantly reduced in the GRP75-siRNA group compared with that in the control (Control: 29.3% vs. GRP75- siRNA (25 nM): 15.7%, p < 0.001). Conclusions: These findings demonstrate that the GRP75-derived MAM region is involved in the development of early embryos in porcine blastocysts.

목차

ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
Chemicals and animals
In vitro maturation (IVM) of oocyte
In vitro fertilization (IVF) and IVC
Transfection of siRNA for porcine BIP/GRP75
Immunofluorescence and Mito-Tracker and ER-TrackerTM staining
Measurement of the mitochondrial Ca2+ level
Protein extraction and western blot analysis
Statistical analysis
RESULTS
Expression of GRP75 and mitochondrial Ca2+ in porcine embryos during IVC
Effects of GRP75 on blastocyst developmental capacity through MAM in porcine embryos
DISCUSSION
CONCLUSION
REFERENCES

키워드

저자

• Hyo-Jin Park [ Department of Biotechnology, Daegu University/DU Center for Infertility, Daegu University, Gyeongsan 38453, Korea ]
• Young-Seo Jo [ Department of Biotechnology, Daegu University/DU Center for Infertility, Daegu University, Gyeongsan 38453, Korea ]
• Seul-Gi Yang [ DU Center for Infertility, Daegu University/Department of Companion Animal Industry, Daegu University, Gyeongsan 38453, Korea ]
• Deog-Bon Koo [ Department of Biotechnology, Daegu University/DU Center for Infertility, Daegu University/Department of Companion Animal Industry, Daegu University, Gyeongsan 38453, Korea ] Corresponding Author

참고문헌

발행기관

간행물

관심저널 등록

• 간행물명

  Journal of Animal Reproduction and Biotechnology

• 간기

  계간

• pISSN

  2671-4639

• eISSN

  2671-4663

• 수록기간

  2019~2026

• 십진분류

  KDC 527 DDC 636