IGF-1 treatment during <i>in vitro</i> maturation improves developmental potential of ovine oocytes through the regulation of PI3K/Akt and apoptosis signaling

Javvaji, Pradeep Krishna; Dhali, A.; Francis, Joseph Rabinson; Kolte, Atul P.; Roy, Sudhir C.; Selvaraju, Sellappan; Mech, A.; Sejian, Veerasamy

doi:10.1080/10495398.2020.1752703

Cited by 18 publications

(16 citation statements)

References 43 publications

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“…5 a), which revealed that the regulatory effects of ARHGAP10 can be partially attributed to the PI3K/Akt/GSK3β signaling pathways. Moreover, to clarify whether the PI3K/Akt signaling pathway directly mediates the effects of ARHGAP10 on cell metastasis, A549 and NCI-H1299 cells treated with or without ARHGAP10 were exposed to insulin-like growth factors-1 (IGF-1) (100 ng/mL), an activator of PI3K/Akt pathway in tumor progression [ 16 , 17 ], and evaluated the expression level of EMT biomarkers by Western blot analysis. The results indicated that IGF-1 (100 ng/mL) could reverse the suppression of EMT via ARHGAP10 treatment (Fig.…”

Section: Resultsmentioning

confidence: 99%

ARHGAP10 inhibits the epithelial–mesenchymal transition of non-small cell lung cancer by inactivating PI3K/Akt/GSK3β signaling pathway

et al. 2021

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Background Rho GTPase activating protein 10 (ARHGAP10) has been implicated as an essential element in multiple cellular process, including cell migration, adhesion and actin cytoskeleton dynamic reorganization. However, the correlation of ARHGAP10 expression with epithelial–mesenchymal transition (EMT) in lung cancer cells is unclear and remains to be elucidated. Herein, we investigated the relationship between the trait of ARHGAP10 and non-small cell lung cancer (NSCLC) pathological process. Methods Immunohistochemistry was conducted to evaluate the expression of ARHGAP10 in NSCLC tissues. CCK-8 assays, Transwell assays, scratch assays were applied to assess cell proliferation, invasion and migration. The expression levels of EMT biomarkers and active molecules involved in PI3K/Akt/GSK3β signaling pathway were examined through immunofluorescence and Western blot. Results ARHGAP10 was detected to be lower expression in NSCLC tissues compared with normal tissues from individuals. Moreover, overexpression of ARHGAP10 inhibited migratory and invasive potentials of A549 and NCI-H1299 cells. In addition, ARHGAP10 directly mediated the process of EMT via PI3K/Akt/GSK3β pathway. Meanwhile, activation of the signaling pathway of insulin-like growth factors-1 (IGF-1) reversed ARHGAP10 overexpression regulated EMT in NSCLC cells. Conclusion ARHGAP10 inhibits the epithelial–mesenchymal transition in NSCLC via PI3K/Akt/GSK3β signaling pathway, suggesting agonist of ARHGAP10 may be an optional remedy for NSCLC patients than traditional opioids.

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Section: Resultsmentioning

confidence: 99%

ARHGAP10 inhibits the epithelial–mesenchymal transition of non-small cell lung cancer by inactivating PI3K/Akt/GSK3β signaling pathway

et al. 2021

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“…Culture conditions such as media composition and presence of growth factors are critical to oocyte development during IVM, and subsequent embryo development competence [ 34 , 35 ]. In recent years, many studies have been attempted to establish the optimal condition of IVM by supplementation of growth factors [ 36 , 37 ], extracellular vesicles [ 38 ], follicular fluid [ 5 ], and mimicking the in vivo microenvironment using a coculture system with fresh oocyte [ 39 ], denuded oocyte [ 40 ], and somatic cells such as cumulus cells [ 41 ] and oviduct cells [ 42 , 43 ]. As the coculture system provides COCs with a similar in vivo microenvironment as closely as possible by transporting multiple paracrine factors into the culture medium [ 13 ], the secreted factors derived from the conditioned medium can positively effect on in vitro oocyte development, which is validated by our findings.…”

Section: Discussionmentioning

confidence: 99%

Human Adipose-Derived Stem Cells’ Paracrine Factors in Conditioned Medium Can Enhance Porcine Oocyte Maturation and Subsequent Embryo Development

Lee

2021

IJMS

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An essential requirement for the success of in vitro maturation (IVM) of the oocyte is to provide an optimal microenvironment similar to in vivo conditions. Recently, somatic cell-based coculture or supplementation of a conditioned medium during IVM has been performed to obtain better quality of oocytes, because they mimic the in vivo reproductive tract by secreting paracrine factors. In this study, human adipose-derived stem cells (ASC) and their conditioned medium (ASC-CM) were applied to IVM of porcine oocytes to evaluate the effectiveness of ASC on oocyte development and subsequent embryo development. In results, both ASC and ASC-CM positively influence on oocyte maturation and embryo development by regulating growth factor receptors (VEGF, FGFR, and IGFR), apoptosis (BCL2), cumulus expansion (PTGS2, HAS2, and TNFAIP6), and oocyte maturation-related genes (GDF9 and BMP15). In particular, the fluorescence intensity of GDF9 and BMP15 was markedly upregulated in the oocytes from the ASC-CM group. Furthermore, significantly high levels of growth factors/cytokine including VEGF, bFGF, IGF-1, IL-10, and EGF were observed in ASC-CM. Additionally, the ASC-CM showed active scavenging activity by reducing the ROS production in a culture medium. Consequently, for the first time, this study demonstrated the effect of human ASC-CM on porcine oocyte development and the alteration of mRNA transcript levels in cumulus–oocyte complexes.

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“…Ovine COC were collected and subjected to IVM as described previously ( Javvaji et al, 2020 ). Briefly, ovine ovaries were collected from a local abattoir and COC were aspirated from the 2–6 mm follicles in aspiration medium (HEPES-buffered M199 supplemented with 50 IU/ml heparin, 50 μg/ml gentamicin and 4 mg/ml fatty acid free BSA fraction V).…”

Section: Methodsmentioning

confidence: 99%

“…Ovine embryos were produced in vitro according to the methods described previously (Javvaji et al, 2020). Briefly, COC were matured in vitro as described above without any supplementation.…”

Section: In Vitro Embryo Productionmentioning

confidence: 99%

An Efficient Nitroblue Tetrazolium Staining and Bright-Field Microscopy Based Method for Detecting and Quantifying Intracellular Reactive Oxygen Species in Oocytes, Cumulus Cells and Embryos

Javvaji

Dhali

Francis

et al. 2020

Front. Cell Dev. Biol.

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Assessment of intracellular reactive oxygen species (ROS) is important for evaluating the developmental ability of cumulus-oocyte complexes (COC) and embryos. Although, fluorescence-based 2 ,7-dichlorodihydrofluorescein diacetate (DCFH-DA) staining method is used widely for detecting intracellular ROS in COC and embryos, it is associated with several limitations. This study aimed to develop an alternative method for detecting and quantifying intracellular ROS in oocytes, cumulus cells and embryos based on nitroblue tetrazolium (NBT) staining and bright-field microscopy. Nitroblue tetrazolium reacts with ROS and forms formazan precipitate that can be detected as dark purple/blue spots under bright-field microscope. Ovine COC were matured in vitro without (control) or with the supplementation of Interleukin-7 (IL-7; for stimulating intracellular ROS), Tempol (superoxide scavenger) or combination of IL-7 and Tempol. The matured COC were stained with NBT and the formation of intracellular formazan precipitates was assessed. Additionally, the matured COC were stained with DCFH-DA to compare the level of intracellular ROS. Further, ovine embryos (8-cell, morula, and degenerating) were generated in vitro and stained with NBT for assessing intracellular ROS. The level of intracellular ROS was expressed as the proportion (%) of the NBT stained area of oocytes, compact cumulus cell masses or embryos. The proportions of NBT stained area in the matured oocytes and cumulus cells was found significantly lesser in the control as compared to the IL-7 (1 and 5 ng/ml) treated groups. A similar trend in the intracellular ROS level was also observed in the matured COC, when assessed based on the DCFH-DA staining. Following the treatment with Tempol (100 mM), negligible NBT stained area in oocytes and cumulus cells was observed. The NBT staining patterns of the oocytes and cumulus cells following the combined

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IGF-1 treatment during in vitro maturation improves developmental potential of ovine oocytes through the regulation of PI3K/Akt and apoptosis signaling

Cited by 18 publications

References 43 publications

ARHGAP10 inhibits the epithelial–mesenchymal transition of non-small cell lung cancer by inactivating PI3K/Akt/GSK3β signaling pathway

ARHGAP10 inhibits the epithelial–mesenchymal transition of non-small cell lung cancer by inactivating PI3K/Akt/GSK3β signaling pathway

Human Adipose-Derived Stem Cells’ Paracrine Factors in Conditioned Medium Can Enhance Porcine Oocyte Maturation and Subsequent Embryo Development

An Efficient Nitroblue Tetrazolium Staining and Bright-Field Microscopy Based Method for Detecting and Quantifying Intracellular Reactive Oxygen Species in Oocytes, Cumulus Cells and Embryos

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