Distinct roles of Cep192 and Cep152 in acentriolar MTOCs and spindle formation during mouse oocyte maturation

Lee, Inwon; Jo, Yu-Jin; Jung, Seung-Min; Wang, Hai Yang; Kim, Nam‐Hyung; Namgoong, Suk

doi:10.1096/fj.201700559rr

Cited by 19 publications

(26 citation statements)

References 44 publications

(83 reference statements)

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“…To assess the involvement of TP53BP1 in the reorganization of MTOCs in more detail, we monitored the reorganization of Cep192, a MTOC marker, in TP53BP1‐knockdown oocytes (Clift & Schuh, ; Lee et al, ). Live‐cell imaging was used to monitor the control and TP53BP1‐knockdown oocytes by labeling the microtubules with SiR‐tubulin (Lukinavicius et al, ) and injecting them with GFP‐Cep192 (Lee et al, ) In the control oocytes, we observed typical patterns of MTOC clustering and bipolar spindle assembly, as reported previously (Figure a, Movie S1; Schuh & Ellenberg, ); during spindle elongation in prometaphase, MTOCs were localized at both ends of the spindle poles (5 hr after GVBD). In contrast, the TP53BP1‐knockdown oocytes showed several MTOC foci remaining at the center of the spindle axis (Figure a, Movies S2‐S4), indicating that the TP53BP1 knockdown causes defects in MTOC localization and stability after prometaphase I.…”

Section: Resultsmentioning

confidence: 99%

“…Oocytes, that undergo meiosis, lack the classic centrosome structure and instead contain microtubule‐organizing centers (MTOCs), which act as centers for microtubule nucleation (Clift & Schuh, ; Schatten & Sun, ). Mouse oocytes lack centrioles but contain PCM proteins including Cep192 and Cep152 (Lee et al, ), CIP2A (Wang et al, ), Polo‐like kinase 1 (PLK1; Solc et al, ; Tong et al, ), and Aurora Kinase A (AURKA; Shuda, Schindler, Ma, Schultz, & Donovan, ). In mouse oocytes, spindle assembly starts after germinal vesicle breakdown (GVBD) by MTOCs, which forms cytoplasmic microtubule networks.…”

Section: Introductionmentioning

confidence: 99%

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TP53BP1 regulates chromosome alignment and spindle bipolarity in mouse oocytes

Jin

Namgoong

2019

Molecular Reproduction Devel

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Meiotic oocytes lack classic centrosomes; therefore, bipolar spindle assembly depends on the clustering of acentriolar microtubule‐organizing centers (MTOCs) into two poles. The bipolar spindle is an essential cellular component that ensures accurate chromosome segregation during anaphase. If the spindle does not form properly, it can result in aneuploidy or cell death. However, the molecular mechanism by which the bipolar spindle is established is not yet fully understood. Tumor suppressor p53‐binding protein 1 (TP53BP1) is known to mediate the DNA damage response. Several recent studies have indicated that TP53BP1 has noncanonical roles in processes, such as spindle formation; however, the role of TP53BP1 in oocyte meiosis is currently unclear. Our results show that TP53BP1 knockdown affects spindle bipolarity and chromatin alignment by altering MTOC stability during oocyte maturation. TP53BP1 was localized in the cytoplasm and displayed an irregular cloud pattern around the spindle/chromosome region. TP53BP1 was also required for the correct localization of MTOCs into the two spindle poles during pro‐meiosis I. TP53BP1 deletion altered the MTOC‐localized Aurora Kinase A. TP53BP1 knockdown caused the microtubules to detach from the kinetochores and increased the rate of aneuploidy. Taken together, our data show that TP53BP1 plays crucial roles in chromosome stability and spindle bipolarity during meiotic maturation.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

TP53BP1 regulates chromosome alignment and spindle bipolarity in mouse oocytes

Jin

Namgoong

2019

Molecular Reproduction Devel

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“…Control oocytes were microinjected with green fluorescent protein (GFP) dsRNA. To generate GFP‐Cep192 to confirm Cep192 localization, pRN‐GFP‐Cep192 plasmids (I. W. Lee et al, ) were linearized by digestion with Sfi1, transcribed using the T3 mMessage mMachine Kit (Ambion), and purified by phenol–chloroform and ethanol precipitation. To generate Ran T24N and Ran Q69L complementary RNAs (cRNAs), human Ran Q69L (Addgene plasmid #30309; provided as a gift from Jay Brenman, University of North Carolina) and Ran T24N (Addgene plasmid #37396; provided as a gift from Iain Cheeseman, Whitehead Institute, MIT) were cloned into pRN3 vector (Lemaire, Garrett, & Gurdon, ) as mScarlet fusion, and in vitro transcriptions were performed in the same manner as for Cep192 cRNAs.…”

Section: Methodsmentioning

confidence: 99%

“…The composition and roles of acentriolar MTOCs in the formation of meiotic spindles have been studied in mouse oocytes as a model system (Baumann, Wang, Yang, & Viveiros, ; I. W. Lee et al, ; H. Wang et al, ). In mouse oocytes, acentriolar MTOCs are composed of several centriolar proteins, including pericentrin (PCNT), Cep192, and Cep152 (Clift & Schuh, ; I. W. Lee et al, ; Ma & Viveiros, ), and depletion of these components impairs spindle formation. Human oocytes obtained from infertile patients do not have functional MTOCs, and spindle formation relies on the Ran‐dependent pathways of microtubule nucleation (Holubcova, Blayney, Elder, & Schuh, ).…”

Section: Introductionmentioning

confidence: 99%

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Acentriolar microtubule organization centers and Ran‐mediated microtubule formation pathways are both required in porcine oocytes

Kim

Lee

Jin

et al. 2019

Molecular Reproduction Devel

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Mammalian oocytes lack centrioles but can generate bipolar spindles using several different mechanisms. For example, mouse oocytes have acentriolar microtubule organization centers (MTOCs) that contain many components of the centrosome, and which initiate microtubule polymerization. On the contrary, human oocytes lack MTOCs and the Ran‐mediated mechanisms may be responsible for spindle assembly. Complete knowledge of the different mechanisms of spindle assembly is lacking in various mammalian oocytes. In this study, we demonstrate that both MTOC‐ and Ran‐mediated microtubule nucleation are required for functional meiotic metaphase I spindle generation in porcine oocytes. Acentriolar MTOC components, including Cep192 and pericentrin, were absent in the germinal vesicle and germinal vesicle breakdown stages. However, they start to colocalize to the spindle microtubules, but are absent in the meiotic spindle poles. Knockdown of Cep192 or inhibition of Polo‐like kinase 1 activity impaired the recruitment of Cep192 and pericentrin to the spindles, impaired microtubule assembly, and decreased the polar body extrusion rate. When the RanGTP gradient was perturbed by the expression of dominant negative or constitutively active Ran mutants, severe defects in microtubule nucleation and cytokinesis were observed, and the localization of MTOC materials in the spindles was abolished. These results demonstrate that the stepwise involvement of MTOC‐ and Ran‐mediated microtubule assembly is crucial for the formation of meiotic spindles in porcine oocytes, indicating the diversity of spindle formation mechanisms among mammalian oocytes.

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Perfluorooctanoic acid exposure in vivo perturbs mitochondrial metabolic during oocyte maturation

Zhang

et al. 2022

Environmental Toxicology

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Perfluorooctanoic acid (PFOA), a member of a group of polyfluorinated and perfluorinated alkyl substances (PFAS), is associated with adverse pregnancy outcomes in mammals. However, the effects of in vivo exposure to PFOA on the female reproductive system and the underlying mechanisms remain unclear. In our study, we constructed a mouse model to investigate whether low-dose PFOA (1 mg/kg/day) or high-dose PFOA (5 mg/kg/day) affect meiosis maturation of oocytes and the potential mechanisms that may be associated with oocyte maturation disorder. Our results indicate that low-dose and high-dose PFOA can lead to impaired oocyte maturation, which is manifested by decreased rate of embryonic foam rupture and first polar body extrusion. Moreover, PFOA exposure harmed the mitochondrial metabolic, resulting in low levels of ATP contents, high reactive oxygen species, aberrant mitochondrial membrane potential. In addition, the proportion of DNA damage marker γ-H2AX was also significantly increased in PFOA exposure oocytes. These changes lead to abnormal arrangements of the spindle and chromosomes during oocyte maturation. In conclusion, our results for the first time illustrated that exposure to PFOA in vivo in female mice impaired the meiosis maturation of oocytes, which provided a basis for studying the mechanism of PFOA reproductive toxicity in female mammals.

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Distinct roles of Cep192 and Cep152 in acentriolar MTOCs and spindle formation during mouse oocyte maturation

Cited by 19 publications

References 44 publications

TP53BP1 regulates chromosome alignment and spindle bipolarity in mouse oocytes

TP53BP1 regulates chromosome alignment and spindle bipolarity in mouse oocytes

Acentriolar microtubule organization centers and Ran‐mediated microtubule formation pathways are both required in porcine oocytes

Perfluorooctanoic acid exposure in vivo perturbs mitochondrial metabolic during oocyte maturation

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