Abstract:LC3 − the mammalian homolog of Atg8 − was found as autophagosome membrane binding protein
in mammals and widely used as an autophagosomal marker. LC3A, B and C show different
expression patterns in each tissue. The aim of this study was to reveal the differences of
expression patterns among LC3 families in mouse placenta under normal condition and
nutrient starving condition. LC3A and B were highly expressed in decidual cells. LC3A and
B were increased in D14 compared with D12 and D16 in mouse placenta, while … Show more
“…S5). This is consistent with the common view that LC3s and GABARAPs have concerted but also can exhibit distinct, non-overlapping intracellular localizations, which is accompanied with at least partial functional divergences 2,16,22,24,25 .Figure 5Anti-GABARAP (8H5) antibody detects endogenous GABARAP within some but not all LC3B + structures. ( A ) HAP1 parental and GABARAP KO cells were incubated for 3 h with 100 nM BafA1 in growth factor depleted medium.…”
The determination of unique functions of GABARAP (gamma-aminobutyric acid type A receptor-associated protein), a member of the highly conserved protein family of mammalian autophagy-related 8 protein (mATG8), within diverse cellular processes remains challenging. Because available anti-GABARAP antibodies perform inadequate, especially within various microscopy-based applications, we aimed to develop an antibody that targets GABARAP but not its close orthologs. Following the latest recommendations for antibody validation including fluorescence protein tagging, genetic and orthogonal strategies, we characterized the resulting anti-GABARAP (8H5) antibody during confocal immunofluorescence imaging in-depth. We compared the antibody staining pattern with that obtained for fluorescence protein tagged GABARAP, GABARAPL1 or GABARAPL2 each ectopically expressed in GABARAP knockout cells. Furthermore, we imaged cells expressing all mATG8 family members at endogenous levels and checked GABARAP knockout cells for unspecific staining under fed or macroautophagy-inducing conditions. Finally, we simultaneously stained cells for endogenous GABARAP and the common autophagosomal marker LC3B. Summarized, the presented antibody shows high specificity for GABARAP without cross-reactivity to other mATG8 family members in immunofluorescence imaging making it a valuable tool for the identification of unique GABARAP functions.
“…S5). This is consistent with the common view that LC3s and GABARAPs have concerted but also can exhibit distinct, non-overlapping intracellular localizations, which is accompanied with at least partial functional divergences 2,16,22,24,25 .Figure 5Anti-GABARAP (8H5) antibody detects endogenous GABARAP within some but not all LC3B + structures. ( A ) HAP1 parental and GABARAP KO cells were incubated for 3 h with 100 nM BafA1 in growth factor depleted medium.…”
The determination of unique functions of GABARAP (gamma-aminobutyric acid type A receptor-associated protein), a member of the highly conserved protein family of mammalian autophagy-related 8 protein (mATG8), within diverse cellular processes remains challenging. Because available anti-GABARAP antibodies perform inadequate, especially within various microscopy-based applications, we aimed to develop an antibody that targets GABARAP but not its close orthologs. Following the latest recommendations for antibody validation including fluorescence protein tagging, genetic and orthogonal strategies, we characterized the resulting anti-GABARAP (8H5) antibody during confocal immunofluorescence imaging in-depth. We compared the antibody staining pattern with that obtained for fluorescence protein tagged GABARAP, GABARAPL1 or GABARAPL2 each ectopically expressed in GABARAP knockout cells. Furthermore, we imaged cells expressing all mATG8 family members at endogenous levels and checked GABARAP knockout cells for unspecific staining under fed or macroautophagy-inducing conditions. Finally, we simultaneously stained cells for endogenous GABARAP and the common autophagosomal marker LC3B. Summarized, the presented antibody shows high specificity for GABARAP without cross-reactivity to other mATG8 family members in immunofluorescence imaging making it a valuable tool for the identification of unique GABARAP functions.
“…Impairment of autophagy by soluble endoglin weakens the invasion and vascular remodelling ability of EVTs, leading to poor placentation. In support of these findings, another study found that autophagy is highly induced in maternal decidual cells and the placenta (Hiyama et al ., ).…”
Section: The Roles Of Autophagy In the Mammalian Life Cyclementioning
Autophagy is primarily an efficient intracellular catabolic pathway used for degradation of abnormal cellular protein aggregates and damaged organelles. Although autophagy was initially proposed to be a cellular stress responder, increasing evidence suggests that it carries out normal physiological roles in multiple biological processes. To date, autophagy has been identified in most organs and at many different developmental stages, indicating that it is not only essential for cellular homeostasis and renovation, but is also important for organ development. Herein, we summarize our current understanding of the functions of autophagy (which here refers to macroautophagy) in the mammalian life cycle.
“…The expression levels of LC3A and LC3B, but not LC3C, are higher in the decidua basalis than in the labyrinth layer. 30 LC3B expression primarily increases in the labyrinth and decidua basalis in response to starvation, which is a general inducer of autophagy in the murine placenta. Although the underlying mechanisms by which starvation increases LC3B expression are not known, LC3B could be responsible for autophagy response in the murine placenta.…”
Section: Role Of Autophagy In In Vivo Embryogenesis and Placentationmentioning
confidence: 99%
“…The LC3 family of proteins is divided into three types: LC3A, LC3B and LC3C. The expression levels of LC3A and LC3B, but not LC3C, are higher in the decidua basalis than in the labyrinth layer 30 . LC3B expression primarily increases in the labyrinth and decidua basalis in response to starvation, which is a general inducer of autophagy in the murine placenta.…”
Section: Role Of Autophagy In In Vivo Embryogenesis and Placentationmentioning
Hypertensive disorders of pregnancy, including preeclampsia, directly affect maternal and perinatal morbidity and mortality. As the pathophysiology of preeclampsia is multi-factorial and has been studied using different approaches, we have demonstrated that impaired autophagy is an intertwined risk factor for preeclampsia. This concept has been verified in both in vitro and in vivo experiments. Autophagy is primarily involved in maintaining cellular homeostasis, and in immune regulation, longevity, cytokines secretion and a variety of other biological functions. Here, we review the role of autophagy in normal embryogenesis and placentation. Once placental autophagy is impaired by metabolic stress such as hypoxia, endoplasmic reticulum stress or starvation, placental development could be disrupted, resulting in functional maladaptations at the maternal-fetal interface. These malfunctions may result in fetal growth restriction or preeclampsia.
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