The regulation of autophagy in porcine blastocysts: Regulation of PARylation-mediated autophagy via mammalian target of rapamycin complex 1 (mTORC1) signaling
| Autor: | Hoon Taek Lee, Jun Sung Lee, Min Gyeong Kim, Hye Ran Lee, Jeong Ho Hwang, Duk Hyoun Kim |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Swine Poly ADP ribose polymerase Biophysics mTORC1 Mechanistic Target of Rapamycin Complex 1 Biology Biochemistry 03 medical and health sciences Autophagy medicine Animals Blastocyst Molecular Biology Cells Cultured PI3K/AKT/mTOR pathway Sirolimus TOR Serine-Threonine Kinases Gene Expression Regulation Developmental Cell Biology Molecular biology 030104 developmental biology medicine.anatomical_structure Apoptosis Multiprotein Complexes Benzamides embryonic structures Phosphorylation Poly(ADP-ribose) Polymerases |
| Zdroj: | Biochemical and Biophysical Research Communications. 473:899-906 |
| ISSN: | 0006-291X |
| DOI: | 10.1016/j.bbrc.2016.03.148 |
| Popis: | Poly(ADP-ribosyl)ation (PARylation) acts as a modulator of selective autophagic degradation of ubiquitinated aggregates for cellular quality control, functioning in pro-survival role. It was reported previously that the inhibition of PARylation resulted in autophagy defects leading accumulation of ubiquitinated aggregates SQSTM1/p62 and apoptosis in porcine blastocysts. Thus, this study aims to investigate the mechanism between PARylation and autophagy in porcine blastocysts. In vitro produced (IVP) embryos were treated with 3-aminobenzamide (3ABA, poly (ADP-ribose) polymerase inhibitor) and/or rapamycin (RAPA, an mTORC1 inhibitor) during blastocyst formation. Then, these treated blastocysts were analyzed by real-time PCR, immunocytochemistry and TUNEL Assay. We found that the 3ABA treatment increased mTORC1 downstream target, phosphorylation of thr389 p70S6K (p-p70S6K-thr389), suggesting an increase in mTORC1 activity. Co-treatment with rapamycin (RAPA), mTORC1 inhibitor, restored the 3ABA-induced autophagy defects to those of the controls by normalizing mTORC1 activity. Moreover, autophagy induction, with only RAPA treatment, increased the rate of blastocyst development (70.05 ± 0.93 vs. 50.61 ± 3.49%), total cell number (58.48 ± 2.94 vs. 49.58 ± 2.43) and blastomere survival, but decreased the accumulation of SQSTM1/p62 aggregates. In summary, mTORC1 signaling is a key mechanism of PARylation-autophagy and its inhibition improved developmental ability and embryo quality by promoting selective autophagic degradation of ubiquitinated aggregates in porcine blastocysts. Therefore, these findings have significant implications for understanding the importance of autophagy regulation for successful in vitro production of porcine embryos. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect