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在山羊单细胞期体细胞核移植胚胎中诱导自噬可改善植入前胚胎发育。

Induction of autophagy in one-cell stage somatic cell nuclear transfer embryos improves preimplantation embryonic development in goat species.

作者信息

Mahvash Nasrin, Moradi-Hajidavaloo Reza, Jafarpour Farnoosh, Hajian Mehdi, Rahimi Mohsen, Sanei Ata-Abadi Nafiseh, Sadeghi Marjan, Nasr-Esfahani Mohammad Hossein

机构信息

Department of Biology, Faculty of Science and Technology, ACECR Institute of Higher Education (Isfahan), Isfahan, Iran.

Department of Animal Biotechnology, Reproductive Biomedicine Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.

出版信息

PLoS One. 2025 Apr 28;20(4):e0314176. doi: 10.1371/journal.pone.0314176. eCollection 2025.

Abstract

Autophagy is a lysosome-mediated catabolic pathway that is dependent on the mammalian target of rapamycin (mTOR). It plays a crucial role in the degradation of aged organelles and macromolecules. Several studies have explored the role of autophagy in embryonic genome activation and its significance during the early preimplantation development of mammals. In our study, we showed that autophagy is inhibited in one-cell stage SCNT embryos when compared to fertilized counterparts in goats. Notably, we found that 6-DMAP, a kinase inhibitor, reduces the phosphorylation of ERK1/2.This reduction correlates with a decrease in autophagy levels, as indicated by the presence of LC3 puncta in 6-DMAP treated embryos. To address the inhibition of autophagy in goat SCNT embryos, we induced autophagy using Rapamycin at concentrations of 10 and 100 nM for 6 hours, immediately following chemical activation. This induction led to a significant improvement in the development of goat SCNT embryos, as evidenced by an increased blastocyst rate compared to the control group. Our findings suggest that the induction of autophagy during early hours of one-cell stage embryos is critical for pre-implantation development in goat SCNT embryos warrant further investigation. This research opens new avenues for understanding the role of autophagy in embryonic development and its applications in reproductive biotechnology.

摘要

自噬是一种由溶酶体介导的分解代谢途径,其依赖于雷帕霉素的哺乳动物靶点(mTOR)。它在衰老细胞器和大分子的降解中起着关键作用。多项研究探讨了自噬在胚胎基因组激活中的作用及其在哺乳动物植入前早期发育过程中的意义。在我们的研究中,我们发现与山羊体内受精的对应胚胎相比,孤雌激活的单细胞期胚胎中的自噬受到抑制。值得注意的是,我们发现激酶抑制剂6-二甲基氨基嘌呤(6-DMAP)可降低细胞外信号调节激酶1/2(ERK1/2)的磷酸化水平。这种降低与自噬水平的下降相关,6-DMAP处理的胚胎中微管相关蛋白1轻链3(LC3)斑点的存在表明了这一点。为了解决山羊孤雌激活胚胎中自噬的抑制问题,我们在化学激活后立即使用浓度为10和100 nM的雷帕霉素诱导自噬6小时。这种诱导导致山羊孤雌激活胚胎的发育有显著改善,与对照组相比,囊胚率增加证明了这一点。我们的研究结果表明,在单细胞期胚胎的早期诱导自噬对于山羊孤雌激活胚胎的植入前发育至关重要,值得进一步研究。这项研究为理解自噬在胚胎发育中的作用及其在生殖生物技术中的应用开辟了新途径。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1cb0/12036934/2d4adb05b8c0/pone.0314176.g001.jpg

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