Autophagy Regulates Embryonic Survival During Delayed Implantation

Lee, Jong-Eun; Oh, Hyun-Ah; Song, Haengseok; Jun, Jin Hyun; Roh, Cheong‐Rae; Xie, Hua; Dey, Sudhansu K.; Lim, Hyunjung Jade

doi:10.1210/en.2010-1456

Cited by 90 publications

(103 citation statements)

References 36 publications

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“…5A. This pattern is similar to the punctate pattern exhibited in blastocysts obtained from GFP-LC3 transgenic mice (Lee et al 2011). We also confirmed by western blotting that the internalized LDP12-EGFP-LC3 fusion proteins undergo PE conjugation and produce LC3-II forms.…”

Section: Ldp12-mediated Protein Transduction In Blastocystssupporting

confidence: 85%

Application of a novel cell-permeable peptide-driven protein delivery in mouse blastocysts

et al. 2013

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Cell-permeable peptides (CPPs) mediate the delivery of macromolecules into cells. However, whether CPPs are usable in mammalian oocytes and embryos for the modulation of protein expression has not been widely investigated. We have previously designed a novel 12-mer CPP from the conserved region of the human papillomavirus L1 capsid protein.In this study, we tested whether this peptide, LDP12, effectively delivers a protein cargo to mouse oocytes and preimplantation embryos. We prepared a LDP12-EGFP fusion protein having LDP12 as an N-terminal tag. This fusion protein readily enters HeLa cells, a cervical cancer cell line. The entry of LDP12-EGFP was partially blocked by amiloride, while cytochalasin D or methyl-b-cyclodextrin slightly increased the uptake. LDP12-EGFP shows efficient transduction in mouse blastocysts, but not in oocytes, two-cell-stage, or morula-stage-preimplantation embryos. LDP12-mediated delivery of EGFP-LC3, a widely used marker of autophagic activation, is successful in HeLa cells and mouse blastocysts, as it enters cells and exhibits a signature punctate pattern. The lipidation of EGFP-LC3 also normally occurs after transduction, suggesting that the transduced protein retains the functional characteristics. Collectively, we show that LDP12-driven protein delivery is a fast and convenient method applicable to mouse blastocysts and reproductive cancer cells.

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Section: Ldp12-mediated Protein Transduction In Blastocystssupporting

confidence: 85%

Application of a novel cell-permeable peptide-driven protein delivery in mouse blastocysts

et al. 2013

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“…By immunofluorescence staining, LC3-II form appears as puncta in the cytoplasm, indicating their accumulation in autophagosomes and autophagolysosomes (Lee et al 2011). As shown in Fig.…”

Section: Resultsmentioning

confidence: 67%

“…We previously showed that mouse blastocysts show heightened autophagy when implantation is delayed by ovariectomy (OVX) on day 4 of pregnancy (Lee et al 2011). Under conditions of estrogen deprivation, dormant blastocysts survive in utero while the uterine quiescence is maintained by daily P 4 injection (Dey & Lim 2006).…”

Section: Introductionmentioning

confidence: 99%

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Suppression of autophagic activation in the mouse uterus by estrogen and progesterone

Choi¹,

Shin²,

Song

et al. 2014

Journal of Endocrinology

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Autophagy is a major cellular catabolic pathway tightly associated with cell survival. The involvement of autophagy in the prolonged survival of blastocysts in the uterus is well established, and it was assumed that ovarian steroid hormones -progesterone (P 4 ) and estrogens -have important roles in the regulation of autophagy. However, information is scarce regarding whether these hormones regulate autophagy in certain hormoneresponsive cellular systems. In this study, we investigated the effects of estrogen and P 4 on autophagic response in the uteri of pregnant mice and in ovariectomized (OVX) mice treated with hormones. During pregnancy, autophagic response is high on days 1 and 2 when the uterus shows an inflammatory response to mating, but it subsides around the time of implantation. Dexamethasone treatment to day 1 pregnant mice reduced autophagy in the uterus. In OVX mouse uteri, estrogen or P 4 reduces autophagic response within 6 h. Glycogen content in OVX uteri was increased by 3-methyladenine treatment, suggesting that autophagy is involved in glycogen breakdown in the hormone-deprived uterus. The classical nuclear receptor antagonists, ICI 182 780 or mifepristone, lead to the recovery of the autophagic response in OVX uteri. The suppression of autophagy by 17b-estradiol is inversely correlated with the accumulation of phospho-mouse target of rapamycin, and rapamycin treatment is moderately effective in the upregulation of autophagic response in OVX mouse uteri. Collectively, this study establishes that the uterine autophagy is induced in hormone-derived environment and is suppressed by hormone treatment. Uterine autophagy may have multiple functions as a responsive mechanism to acute inflammation and as an energy provider by breaking down glycogen under hormone deprivation.

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“…10 It was also hypothesized that autophagy in trophectoderm might be a surviving mechanism of dormant blastocysts in an experimentally delayed implantation model. 11 These findings imply that autophagy is involved in the placental pathophysiology in addition to apoptosis.…”

Section: Introductionmentioning

confidence: 95%

Autophagy Induced by Tumor Necrosis Factor α Mediates Intrinsic Apoptosis in Trophoblastic Cells

et al. 2014

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To investigate the interconnection of apoptosis and autophagy in trophoblastic cells, we treated JEG-3 cells with tumor necrosis factor a (TNF-a) after transfecting LC3 or Beclin 1 or calpain small interfering RNA (siRNA), which blocks cleavage of autophagyrelated gene 5 (Atg5) into N-terminal truncated Atg5 (tAtg5), a mediator between apoptosis and autophagy, and assessed the changes in LC3-II, caspase 9, caspase 3, and tAtg5. We also assessed the TNF-a-induced changes in LC3-II, caspase 9, and caspase 3 in primary trophoblasts from term placentae after transfecting siRNA for LC3 or Beclin 1. In both types of cells, transfection of LC3 or Beclin 1 siRNA significantly attenuated TNF-a-induced increases in LC3-II and activations of caspase 9 and caspase 3. There was significant abrogation of TNF-a-induced expression of tAtg5 after transfection with LC3 or Beclin 1 siRNA. Moreover, transfection with calpain siRNA significantly decreased TNF-a-induced changes in caspase 3 and caspase 9 in addition to tAtg5 in JEG-3 cells. Our data suggest that TNF-a-induced autophagy mediates intrinsic apoptosis, probably through tAtg5, in trophoblastic cells.

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Autophagy Regulates Embryonic Survival During Delayed Implantation

Cited by 90 publications

References 36 publications

Application of a novel cell-permeable peptide-driven protein delivery in mouse blastocysts

Application of a novel cell-permeable peptide-driven protein delivery in mouse blastocysts

Suppression of autophagic activation in the mouse uterus by estrogen and progesterone

Autophagy Induced by Tumor Necrosis Factor α Mediates Intrinsic Apoptosis in Trophoblastic Cells

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