Microtubule organizing centers regulate spindle positioning in mouse oocytes

Londoño-Vásquez, Daniela; Rodriguez-Lukey, Katherine; Behura, Susanta K.; Balboula, Ahmed Z.

doi:10.1016/j.devcel.2021.12.011

Cited by 19 publications

(19 citation statements)

References 55 publications

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“…Germinal vesicle oocytes were cultured in pre‐warmed and equilibrated maturation medium and imaged over time under a 40× immersion oil objective using a Leica TCS SP8 confocal microscope equipped with a microenvironmental chamber to regulate the temperature and CO 2 at 37°C and 5% in humidified air. SiR‐tubulin (Cytoskeleton NC0958386) was added to the maturation medium to label microtubules 30,31 . Bright‐field and 647 nm wavelength images acquisition were started at 1.5 h after collection (30–45 min collection time), in which the oocytes were at the GVBD stage.…”

Section: Methodsmentioning

confidence: 99%

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Adverse PFAS effects on mouse oocyte in vitro maturation are associated with carbon‐chain length and inclusion of a sulfonate group

et al. 2022

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Objectives: Per-and polyfluoroalkyl substances (PFAS) are man-made chemicals that are widely used in various products. PFAS are characterized by their fluorinated carbon chains that make them hard to degrade and bioaccumulate in human and animals. Toxicological studies have shown PFAS toxic effects: cytotoxicity, immunotoxicity, neurotoxicity, and reproductive toxicity. However, it is still unclear how the structures of PFAS, such as carbon-chain length and functional groups, determine their reproductive toxicity.Methods and Results: By using a mouse-oocyte-in-vitro-maturation (IVM) system, we found the toxicity of two major categories of PFAS, perfluoroalkyl carboxylic acid (PFCA) and perfluoroalkyl sulfonic acid (PFSA), is elevated with increasing carbonchain length and the inclusion of the sulfonate group. Specifically, at 600 μM, perfluorohexanesulfonic acid (PFHxS) and perfluorooctanesulfonic acid (PFOS) reduced the rates of both germinal-vesicle breakdown (GVBD) and polar-body extrusion (PBE) as well as enlarged polar bodies. However, the shorter PFSA, perfluorobutanesulfonic acid (PFBS), and all PFCA did not show similar adverse cytotoxicity. Further, we found that 600 μM PFHxS and PFOS exposure induced excess reactive oxygen species (ROS) and decreased mitochondrial membrane potential (MMP). Cytoskeleton analysis revealed that PFHxS and PFOS exposure induced chromosome misalignment, abnormal F-actin organization, elongated spindle formation, and symmetric division in the treated oocytes. These meiotic defects compromised oocyte developmental competence after parthenogenetic activation. Conclusions:Our study provides new information on the structure-toxicity relationship of PFAS.

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Section: Methodsmentioning

confidence: 99%

“…SiR‐tubulin (Cytoskeleton NC0958386) was added to the maturation medium to label microtubules. 30 , 31 Bright‐field and 647 nm wavelength images acquisition were started at 1.5 h after collection (30–45 min collection time), in which the oocytes were at the GVBD stage. Time‐lapse images were taken every 30 min.…”

Section: Methodsmentioning

confidence: 99%

Adverse PFAS effects on mouse oocyte in vitro maturation are associated with carbon‐chain length and inclusion of a sulfonate group

et al. 2022

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“…SiR-tubulin (Cytoskeleton NC0958386) was added to the maturation medium to label microtubules. 30,31 Bright-field and 647 nm wavelength images acquisition were started at 1.5 hours after collection (30-45 minutes collection time), in which the oocytes were at the GVBD stage. Time-lapse images were taken every 30 minutes.…”

Section: Methodsmentioning

confidence: 99%

Adverse PFAS effects on mouse oocyte in vitro maturation are associated with carbon-chain length and inclusion of a sulfonate group

Feng

Soto‐Moreno

Prakash

et al. 2022

Preprint

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Per- and polyfluoroalkyl substances (PFAS) are man-made chemicals that are used in products such as non-stick cookware, stain-resistant coating, and food packaging. PFAS are characterized by their fluorinated carbon chains that make them hard to degrade and bioaccumulate in human and animals. Toxicological studies have shown PFAS toxic effects: cytotoxicity, immunotoxicity, neurotoxicity, and reproductive toxicity. Two major categories of PFAS are perfluoroalkyl carboxylic acid (PFCA) and perfluoroalkyl sulfonic acid (PFSA). In this study, we used a mouse-oocyte-in-vitro-maturation (IVM) system to study how the structures of PFAS, such as carbon-chain length and functional groups, determine their reproductive toxicity. We found the toxicity of PFAS is elevated with increasing carbon-chain length and the inclusion of the sulfonate group. Specifically, at 600 μM, perfluorohexanesulfonic acid (PFHxS) and perfluorooctanesulfonic acid (PFOS) reduced the rates of both germinal vesicle breakdown (GVBD) and polar body extrusion (PBE) as well as induced the formation of relatively large polar bodies. However, the shorter PFSA, perfluorobutanesulfonic acid (PFBS), and all PFCA did not show similar adverse cytotoxicity. We further examined mitochondria and cytoskeleton, two essential factors for cell division, in PFOS- and PFHxS-treated oocytes. We found that 600 μM PFHxS and PFOS exposure induced excess reactive oxygen species (ROS) and decreased mitochondrial membrane potential (MMP). Cytoskeleton analysis revealed that PFHxS and PFOS exposure induced chromosome misalignment, abnormal F-actin organization, elongated the spindle formation, and symmetric division in the treated oocytes. Together, our study provides new information on the structure-toxicity relationship of PFAS.

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“…Prophase I-arrested oocytes were cultured in vitro in milrinone-free CZB medium supplemented with 100 nM SiR-tubulin (Cytoskeleton #NC0958386) in a humidified, microenvironmental chamber (5% CO 2 and 37° C) equipped to a Leica TCP SP8 inverted microscope. After culturing cells for 11 hours, mMB caps were partially ablated using a multi-photon laser as previously described 42 . In brief, a 4µm 2 square region of interest within the mMB cap was exposed to a 740 nm wavelength and 60-70 mW power laser beam at the sample plane.…”

Section: Methodsmentioning

confidence: 99%

A meiotic midbody structure in mouse oocytes acts as a barrier for nascent translation to ensure developmental competence

Jung

Londoño-Vásquez

Park

et al. 2022

Preprint

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Successful embryo development is dependent upon maternally deposited components. During egg formation, developmental competence is acquired through regulated translation of maternal mRNA stores. In addition, egg precursors undergo two rounds of chromosome segregation, each coupled to an asymmetric cytokinesis that produces two non-functional polar bodies. In somatic cells, cytokinesis produces two daughter cells and one midbody remnant (MBR), a signaling organelle assembled from the midbody (MB), which first appears in Telophase. MBs contain transcription and translation factors, and epigenetic modifiers. Once MBs mature to MBRs by abscission, they can be subsequently phagocytosed by another cell and influence cellular function or fate. Although the significance of MBs is elucidated in several cell types like neurons, cancer cells and stem cells, the presence and function of MBs in gametes and their roles in reproductive fitness are unknown. Here, we examined the formation and regulation of meiotic midbodies (mMB) in mouse oocytes. We find that although mouse oocyte mMBs contain analogous structures to somatic MBs, they also have a unique cap-like structure composed of the centralspindlin complex, and that cap formation depends upon an asymmetric microtubule abundance in the egg compared to the polar body. Furthermore, our results show that mMBs are translationally active ribonucleoprotein granules, supported by detection of ribosomes, polyadenylated mRNAs and nascent translation. Finally, by pharmacological and laser ablation-based approaches, we demonstrate that the mMB cap is a barrier to prevent translated products from leaving the egg and escaping into the polar body. Crucially, this barrier is critical for successful early embryonic development. Here, we document an evolutionary adaptation to the highly conserved process of cytokinesis in mouse oocytes and describe a new structure and new mechanism by which egg quality and embryonic developmental competence are regulated.

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Microtubule organizing centers regulate spindle positioning in mouse oocytes

Cited by 19 publications

References 55 publications

Adverse PFAS effects on mouse oocyte in vitro maturation are associated with carbon‐chain length and inclusion of a sulfonate group

Adverse PFAS effects on mouse oocyte in vitro maturation are associated with carbon‐chain length and inclusion of a sulfonate group

Adverse PFAS effects on mouse oocyte in vitro maturation are associated with carbon-chain length and inclusion of a sulfonate group

A meiotic midbody structure in mouse oocytes acts as a barrier for nascent translation to ensure developmental competence

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