The subcortical maternal complex protein Nlrp4f is involved in cytoplasmic lattice formation and organelle distribution

Qin, Dandan; Gao, Zheng; Xiao, Yi; Zhang, Xiaoxin; Ma, Hui; Yu, Xingjiang; Nie, Xiaoqing; Fan, Na; Wang, Xiaoqing; Ouyang, Ying‐Chun; Sun, Qing‐Yuan; Yi, Zhaohong; Li, Lei

doi:10.1242/dev.183616

Cited by 25 publications

(18 citation statements)

References 49 publications

(134 reference statements)

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“… (*) Aspect ratio defined according to Russ [ 51 ]. For each condition, data are mean ± standard deviation of n = 50 microbead measures.…”

Section: Resultsmentioning

confidence: 99%

“…ATP6 and ATP8 are mitochondrial genes coding for two subunits of the complex V of the respiratory chain, namely ATP synthase, involved in ATP production [43][44][45]. The subcortical maternal complex (SCMC) is known to play essential roles in early embryo development and female fertility [46][47][48][49][50][51]. It was first described in the subcortex of mouse oocytes and preimplantation embryos as a 669-2000 KD molecular weight complex composed of several proteins encoded by maternal effect genes (MEG) [46].…”

Section: Introductionmentioning

confidence: 99%

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One-step automated bioprinting-based method for cumulus-oocyte complex microencapsulation for 3D in vitro maturation

Mastrorocco¹,

Cacopardo²,

Martino³

et al. 2020

PLoS ONE

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Three-dimensional in vitro maturation (3D IVM) is a promising approach to improve IVM efficiency as it could prevent cumulus-oocyte complex (COC) flattening and preserve its structural and functional integrity. Methods reported to date have low reproducibility and validation studies are limited. In this study, a bioprinting based production process for generating microbeads containing a COC (COC-microbeads) was optimized and its validity tested in a large animal model (sheep). Alginate microbeads were produced and characterized for size, shape and stability under culture conditions. COC encapsulation had high efficiency and reproducibility and cumulus integrity was preserved. COC-microbeads underwent IVM, with COCs cultured in standard 2D IVM as controls. After IVM, oocytes were analyzed for nuclear chromatin configuration, bioenergetic/oxidative status and transcriptional activity of genes biomarker of mitochondrial activity (TFAM, ATP6, ATP8) and oocyte developmental competence (KHDC3, NLRP5, OOEP and TLE6). The 3D system supported oocyte nuclear maturation more efficiently than the 2D control (P<0.05). Ooplasmic mitochondrial activity and reactive oxygen species (ROS) generation ability were increased (P<0.05). Up-regulation of TFAM, ATP6 and ATP8 and down-regulation of KHDC3, NLRP5 expression were observed in 3D IVM. In conclusion, the new bioprinting method for producing COC-microbeads has high reproducibility and efficiency. Moreover, 3D IVM improves oocyte nuclear maturation and relevant parameters of oocyte cytoplasmic maturation and could be used for clinical and toxicological applications.

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“… (*) Aspect ratio defined according to Russ [ 51 ]. For each condition, data are mean ± standard deviation of n = 50 microbead measures.…”

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

One-step automated bioprinting-based method for cumulus-oocyte complex microencapsulation for 3D in vitro maturation

Mastrorocco¹,

Cacopardo²,

Martino³

et al. 2020

PLoS ONE

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“…However, this is a potential limitation of our study that prompts at least two questions: 1) Why was the depletion partial and 2) how could a partial depletion be so detrimental? To answer the first question, we recall that oocytes' structures persisting during preimplantation embryos and even after extraction with detergents have been known for a long time, such as cytoplasmic lattices and SCMC 29,30,61 , which are subcortically located in the ooplasm similar to COPS3. The cortical rim of COPS3 withstood the extraction with Tween 20 and Triton X-100.…”

Section: Discussionmentioning

confidence: 99%

“…Firstly, COPS3 remained present despite chemical extraction. Detergent Triton X-100 is known to extract the majority of oocytic proteins, except for those bound in fibrillar sheets (cytoplasmic lattices) or stable cortical structures [28][29][30] and is harsher than Tween 20. Consistent with the different strengths of the two detergents, the cortical rim of COPS3 withstood better the chemical extraction with 1 % Tween 20 than with 0.1 % Triton X-100 (Figure 3A).…”

Section: Cops3's Cortical Deposit Is Largely Insoluble and Thereby mentioning

confidence: 99%

A Critical Role for The COP9 Signalosome Subunit 3 as A Gatekeeper of Genome Integrity in 2-Cell Mouse Embryos

Israel

Drexler

Fuellen

et al. 2021

Preprint

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Investigations of genes required in early mammalian development are complicated by protein deposits of maternal products, which continue to operate after the gene locus has been disrupted. This leads to an underestimation of the number of genes known to be needed during the embryonic phase of cellular totipotency (up to the 4-cell stage). Here, we expose a critical role of the gene Cops3 by showing that it protects genome integrity during the 2-cell stage of mouse embryo development, in contrast to the previous functional assignment at postimplantation. This new role is mediated by a large, stable and hitherto overlooked deposit of maternal protein. Since protein abundance and stability defeat prospects of DNA- or RNA-based gene inactivation, we adopted a protein strategy of gene inactivation: antigen masking or TRIM21-mediated proteasomal degradation of COPS3. Both resulted in 2-cell embryo lethality, but the expected degradation remained outstanding, because the major fraction of the total COPS3 is secluded in a submembrane cortical rim that withstands extraction with detergent, thereby exposing a soluble vs. insoluble fraction of COPS3, which we ascribe with distinct functional properties. In mechanistic terms, transcriptomic and metabolic analyses reveal that soluble COPS3 is involved in several processes, which, however, converge on DNA endoreduplication and the accumulation of DNA strand breaks in the 2-cell nucleus, where the minor soluble fraction of COPS3 is placed. Thus, we have shown the critical role of maternal protein deposits in development, and we have also highlighted the distinction between the site of accumulation (cell cortex) and point of use (nucleus), which is a dimension of the protein-based strategies of gene inactivation not yet fully appreciated.

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“…To highlight a few, Nlrp2 KO oocytes had decreased developmental potential, and the gene was found to be part of the subcortical maternal complex (SCMC) [38]. Nlrp4f is a gene also associated with the SCMC, and KO mice had decreased fecundity as well as delayed preimplantation development [39]. Finally, Nlrp5 KO oocytes were found to be arrested at the two-cell stage, thus essential for embryonic development [40].…”

Section: Discussionmentioning

confidence: 99%

CRISPR/Cas9-Mediated Genome Editing Reveals Oosp Family Genes are Dispensable for Female Fertility in Mice

Abbasi

Kodani

Emori

et al. 2020

Cells

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There are over 200 genes that are predicted to be solely expressed in the oocyte and ovary, and thousands more that have expression patterns in the female reproductive tract. Unfortunately, many of their physiological functions, such as their roles in oogenesis or fertilization, have yet to be elucidated. Previous knockout (KO) mice studies have proven that many of the genes that were once thought to be essential for fertility are dispensable in vivo. Therefore, it is extremely important to confirm the roles of all genes before spending immense time studying them in vitro. To do this, our laboratory analyzes the functions of ovary and oocyte-enriched genes in vivo through generating CRISPR/Cas9 KO mice and examining their fertility. In this study, we have knocked out three Oosp family genes (Oosp1, Oosp2, and Oosp3) that have expression patterns linked to the female reproductive system and found that the triple KO (TKO) mutant mice generated exhibited decreased prolificacy but were not infertile; thus, these genes may potentially be dispensable for fertility. We also generated Cd160 and Egfl6 KO mice and found these genes are individually dispensable for female fertility. KO mice with no phenotypic data are seldom published, but we believe that this information must be shared to prevent unnecessary experimentation by other laboratories.

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The subcortical maternal complex protein Nlrp4f is involved in cytoplasmic lattice formation and organelle distribution

Cited by 25 publications

References 49 publications

One-step automated bioprinting-based method for cumulus-oocyte complex microencapsulation for 3D in vitro maturation

One-step automated bioprinting-based method for cumulus-oocyte complex microencapsulation for 3D in vitro maturation

A Critical Role for The COP9 Signalosome Subunit 3 as A Gatekeeper of Genome Integrity in 2-Cell Mouse Embryos

CRISPR/Cas9-Mediated Genome Editing Reveals Oosp Family Genes are Dispensable for Female Fertility in Mice

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