Differing molecular response of young and advanced maternal age human oocytes to IVM

Reyes, Juan; Silva, E; Chitwood, James L.; Schoolcraft, W.B.; Krisher, Rebecca L.; Ross, Pablo J.

doi:10.1093/humrep/dex284

Cited by 46 publications

(61 citation statements)

References 43 publications

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“…High throughput techniques are widely applied to understand time-dependent global changes in gene expression during oocyte maturation, oocyte-to-embryo transition and early embryogenic development, often with special attention to human infertility [42]. Even though, methodical improvements of NGS methods have led to the establishment of suitable protocols for transcriptomic studies of mammalian oocytes, no such approach is available for deep-sequencing of polysome-bound mRNAs [55,56]. High quality polysome profiling has conventionally required large number of cells to obtain well-defined and interpretable peaks (e.g., for HeLa cells approximately 2-4 × 10 7 ).…”

Section: Discussionmentioning

confidence: 99%

Identifying the Translatome of Mouse NEBD-Stage Oocytes via SSP-Profiling; A Novel Polysome Fractionation Method

Mašek

Llano

Gahurová

et al. 2020

IJMS

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Meiotic maturation of oocyte relies on pre-synthesised maternal mRNA, the translation of which is highly coordinated in space and time. Here, we provide a detailed polysome profiling protocol that demonstrates a combination of the sucrose gradient ultracentrifugation in small SW55Ti tubes with the qRT-PCR-based quantification of 18S and 28S rRNAs in fractionated polysome profile. This newly optimised method, named Scarce Sample Polysome Profiling (SSP-profiling), is suitable for both scarce and conventional sample sizes and is compatible with downstream RNA-seq to identify polysome associated transcripts. Utilising SSP-profiling we have assayed the translatome of mouse oocytes at the onset of nuclear envelope breakdown (NEBD)—a developmental point, the study of which is important for furthering our understanding of the molecular mechanisms leading to oocyte aneuploidy. Our analyses identified 1847 transcripts with moderate to strong polysome occupancy, including abundantly represented mRNAs encoding mitochondrial and ribosomal proteins, proteasomal components, glycolytic and amino acids synthetic enzymes, proteins involved in cytoskeleton organization plus RNA-binding and translation initiation factors. In addition to transcripts encoding known players of meiotic progression, we also identified several mRNAs encoding proteins of unknown function. Polysome profiles generated using SSP-profiling were more than comparable to those developed using existing conventional approaches, being demonstrably superior in their resolution, reproducibility, versatility, speed of derivation and downstream protocol applicability.

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Section: Discussionmentioning

confidence: 99%

Identifying the Translatome of Mouse NEBD-Stage Oocytes via SSP-Profiling; A Novel Polysome Fractionation Method

Mašek

Llano

Gahurová

et al. 2020

IJMS

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“…Although the current studies were performed in the mouse, TRPM7, CACNA1H, and TRPV3 are all expressed in human oocytes and TRPM7 and TRPV3 are expressed in macaque oocytes as indicated by RNA sequencing (45)(46)(47)(48). Furthermore, there are numerous missense or loss of function (LoF) mutations identified in human TRPM7 (480 missense, 27 LoF), CACNA1H (1,045 missense, 10 LoF), and TRPV3 (312 missense, 23 LoF) (see Exome Aggregation Consortium browser at exac.broadinstitute.org; ref.…”

Section: Discussionmentioning

confidence: 99%

TRPM7 and Ca _V 3.2 channels mediate Ca ²⁺ influx required for egg activation at fertilization

Bernhardt

Stein

Carvacho

et al. 2018

Proc. Natl. Acad. Sci. U.S.A.

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The success of mammalian development following fertilization depends on a series of transient increases in egg cytoplasmic Ca2+, referred to as Ca2+ oscillations. Maintenance of these oscillations requires Ca2+ influx across the plasma membrane, which is mediated in part by T-type, CaV3.2 channels. Here we show using genetic mouse models that TRPM7 channels are required to support this Ca2+ influx. Eggs lacking both TRPM7 and CaV3.2 stop oscillating prematurely, indicating that together they are responsible for the majority of Ca2+ influx immediately following fertilization. Fertilized eggs lacking both channels also frequently display delayed resumption of Ca2+ oscillations, which appears to require sperm–egg fusion. TRPM7 and CaV3.2 channels almost completely account for Ca2+ influx observed following store depletion, a process previously attributed to canonical store-operated Ca2+ entry mediated by STIM/ORAI interactions. TRPM7 serves as a membrane sensor of extracellular Mg2+ and Ca2+ concentrations and mediates the effects of these ions on Ca2+ oscillation frequency. When bred to wild-type males, female mice carrying eggs lacking TRPM7 and CaV3.2 are subfertile, and their offspring have increased variance in postnatal weight. These in vivo findings confirm previous observations linking in vitro experimental alterations in Ca2+ oscillatory patterns with developmental potential and offspring growth. The identification of TRPM7 and CaV3.2 as key mediators of Ca2+ influx following fertilization provides a mechanistic basis for the rational design of culture media that optimize developmental potential in research animals, domestic animals, and humans.

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“…In humans, the oocyte MPF complex is not well characterized [309]. CDK1 [25,79,80], CDC25 [25,84], and WEE2 are expressed in human oocytes [80]. Sang et al found WEE2 mutations in four women with fertilization failure [83].…”

Section: Oocyte Maturation Promoting Factor: Cyclin-dependent Kinase mentioning

confidence: 99%

“…Reyes et al studied molecular responses in 10 oocytes (5 GV, 5 MII) from young women and 10 oocytes (5 GV, 5 MII) from older women using RNA-Seq sequencing (HiSeq 2500; Illumina) [79]. Patients were stimulated with FSH and triggered with HCG.…”

Section: Oocyte Quality and Ivmmentioning

confidence: 99%

Luteinizing Hormone Action in Human Oocyte Maturation and Quality: Signaling Pathways, Regulation, and Clinical Impact

Arroyo

Kim²,

Yeh

2020

Reprod. Sci.

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The ovarian follicle luteinizing hormone (LH) signaling molecules that regulate oocyte meiotic maturation have recently been identified. The LH signal reduces preovulatory follicle cyclic nucleotide levels which releases oocytes from the first meiotic arrest. In the ovarian follicle, the LH signal reduces cyclic nucleotide levels via the CNP/NPR2 system, the EGF/EGF receptor network, and follicle/oocyte gap junctions. In the oocyte, reduced cyclic nucleotide levels activate the maturation promoting factor (MPF). The activated MPF induces chromosome segregation and completion of the first and second meiotic divisions. The purpose of this paper is to present an overview of the current understanding of human LH signaling regulation of oocyte meiotic maturation by identifying and integrating the human studies on this topic. We found 89 human studies in the literature that identified 24 LH follicle/oocyte signaling proteins. These studies show that human oocyte meiotic maturation is regulated by the same proteins that regulate animal oocyte meiotic maturation. We also found that these LH signaling pathway molecules regulate human oocyte quality and subsequent embryo quality. Remarkably, in vitro maturation (IVM) prematuration culture (PMC) protocols that manipulate the LH signaling pathway improve human oocyte quality of cultured human oocytes. This knowledge has improved clinical human IVM efficiency which may become a routine alternative ART for some infertile patients.

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Differing molecular response of young and advanced maternal age human oocytes to IVM

Cited by 46 publications

References 43 publications

Identifying the Translatome of Mouse NEBD-Stage Oocytes via SSP-Profiling; A Novel Polysome Fractionation Method

Identifying the Translatome of Mouse NEBD-Stage Oocytes via SSP-Profiling; A Novel Polysome Fractionation Method

TRPM7 and Ca _V 3.2 channels mediate Ca ²⁺ influx required for egg activation at fertilization

Luteinizing Hormone Action in Human Oocyte Maturation and Quality: Signaling Pathways, Regulation, and Clinical Impact

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Differing molecular response of young and advanced maternal age human oocytes to IVM

Cited by 46 publications

References 43 publications

Identifying the Translatome of Mouse NEBD-Stage Oocytes via SSP-Profiling; A Novel Polysome Fractionation Method

Identifying the Translatome of Mouse NEBD-Stage Oocytes via SSP-Profiling; A Novel Polysome Fractionation Method

TRPM7 and Ca V 3.2 channels mediate Ca 2+ influx required for egg activation at fertilization

Luteinizing Hormone Action in Human Oocyte Maturation and Quality: Signaling Pathways, Regulation, and Clinical Impact

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TRPM7 and Ca _V 3.2 channels mediate Ca ²⁺ influx required for egg activation at fertilization