Morphological and molecular markers are correlated with maturation-competence of human oocytes

Levi, Mattan; Ghetler, Yehudith; Shulman, Adrian; Shalgi, Ruth

doi:10.1093/humrep/det261

Cited by 33 publications

(42 citation statements)

References 55 publications

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“…A direct effect of Fyn on A-type lamins would require the presence of Fyn in the nucleus, which was detected in somatic cells 64, 65 . However, in mouse oocytes Fyn was detected only in the GV of oocytes exhibiting a lower maturation competence 66 . Fyn co-localizes with and phosphorylates β-adducin, an actin filaments associated cytoskeletal protein 24 , supporting the possible action of Fyn on actin filaments.…”

Section: Discussionmentioning

confidence: 94%

Regulation of GVBD in mouse oocytes by miR-125a-3p and Fyn kinase through modulation of actin filaments

Grossman

Har-Paz

Gindi

et al. 2017

Sci Rep

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Meiotically arrested oocytes are characterized by the presence of the nuclear structure known as germinal-vesicle (GV), the breakdown of which (GVBD) is associated with resumption of meiosis. Fyn is a pivotal factor in resumption of the first meiotic division; its inhibition markedly decreases the fraction of oocytes undergoing GVBD. Here, we reveal that in mouse oocytes Fyn is post-transcriptionally regulated by miR-125a-3p. We demonstrate that in oocytes resuming meiosis miR-125a-3p and Fyn exhibit a reciprocal expression pattern; miR-125a-3p decreases alongside with an increase in Fyn expression. Microinjection of miR-125a-3p inhibits GVBD, an effect that is markedly reduced by Fyn over-expression, and impairs the organization of the actin rim surrounding the nucleus. Lower rate of GVBD is also observed in oocytes exposed to cytochalasin-D or blebbistatin, which interfere with actin polymerization and contractility of actin bundles, respectively. By down-regulating Fyn in HEK-293T cells, miR-125a-3p reduces the interaction between actin and A-type lamins, which constitute the nuclear-lamina. Our findings suggest a mechanism, by which a decrease in miR-125a-3p during oocyte maturation facilitates GVBD by allowing Fyn up-regulation and the resulting stabilization of the interaction between actin and A-type lamins.

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Section: Discussionmentioning

confidence: 94%

Regulation of GVBD in mouse oocytes by miR-125a-3p and Fyn kinase through modulation of actin filaments

Grossman

Har-Paz

Gindi

et al. 2017

Sci Rep

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“…GV oocytes were isolated into M2 medium (Sigma) supplemented with 1 μM Milrinon (Sigma) to avoid oocyte maturation. Cumulus cells were removed mechanically, and oocytes were stained as previously described (64). All experimental samples were visualized and photographed using a laser confocal microscope (SP5; Leica) that was calibrated to a secondary-only control.…”

Section: Methodsmentioning

confidence: 99%

Interleukin-1 deficiency prolongs ovarian lifespan in mice

Uri-Belapolsky

Shaish

Eliyahu

et al. 2014

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

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Oocyte endowment dwindles away during prepubertal and adult life until menopause occurs, and apoptosis has been identified as a central mechanism responsible for oocyte elimination. A few recent reports suggest that uncontrolled inflammation may adversely affect ovarian reserve. We tested the possible role of the proinflammatory cytokine IL-1 in the age-related exhaustion of ovarian reserve using IL-1α and IL-1β-KO mice. IL-1α-KO mice showed a substantially higher pregnancy rate and litter size compared with WT mice at advanced age. The number of secondary and antral follicles was significantly higher in 2.5-mo-old IL-1α-KO ovaries compared with WT ovaries. Serum anti-Müllerian hormone, a putative marker of ovarian reserve, was markedly higher in IL-1α-KO mice from 2.5 mo onward, along with a greater ovarian response to gonadotropins. IL-1β-KO mice displayed a comparable but more subtle prolongation of ovarian lifespan compared with IL-1α-KO mice. The protein and mRNA of both IL-1α and IL-1β mice were localized within the developing follicles (oocytes and granulosa cells), and their ovarian mRNA levels increased with age. Molecular analysis revealed decreased apoptotic signaling [higher B-cell lymphoma 2 (BCL-2) and lower BCL-2-associated X protein levels], along with a marked attenuation in the expression of genes coding for the proinflammatory cytokines IL-1β, IL-6, and TNF-α in ovaries of IL-1α-KO mice compared with WT mice. Taken together, IL-1 emerges as an important participant in the age-related exhaustion of ovarian reserve in mice, possibly by enhancing the expression of inflammatory genes and promoting apoptotic pathways.

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“…Importantly, all of these genes are expressed in human oocytes (1, 3, 5). Moreover, both AURKC (34) and FYN (35) have also recently been studied at the protein level in human oocytes.…”

Section: Methodsmentioning

confidence: 99%

Digital multiplexed mRNA analysis of functionally important genes in single human oocytes and correlation of changes in transcript levels with oocyte protein expression

Riris

Webster

Homer

2014

Fertility and Sterility

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ObjectiveTo investigate functionally important transcripts in single human oocytes with the use of NanoString technology and determine whether observed differences are biologically meaningful.DesignAnalysis of human oocytes with the use of NanoString and immunoblotting.SettingUniversity-affiliated reproductive medicine unit.PatientsWomen undergoing in vitro fertilization.InterventionHuman oocytes were analyzed with the use of NanoString or immunoblotting.Main Outcome MeasuresThe abundance of transcripts for ten functionally important genes—AURKA, AURKC, BUB1, BUB1B (encoding BubR1), CDK1, CHEK1, FYN, MOS, MAP2K1, and WEE2—and six functionally dispensable genes were analyzed with the use of NanoString. BubR1 protein levels in oocytes from younger and older women were compared with the use of immunoblotting.Result(s)All ten functional genes but none of the six dispensable genes were detectable with the use of NanoString in single oocytes. There was 3- to 5-fold variation in BUB1, BUB1B, and CDK1 transcript abundance among individual oocytes from a single patient. Transcripts for these three genes—all players within the spindle assembly checkpoint surveillance mechanism for preventing aneuploidy—were reduced in the same oocyte from an older patient. Mean BUB1B transcripts were reduced by 1.5-fold with aging and associated with marked reductions in BubR1 protein levels.Conclusion(s)The abundance of functionally important transcripts exhibit marked oocyte-to-oocyte heterogeneity to a degree that is sufficient to affect protein expression. Observed variations in transcript abundance are therefore likely to be biologically meaningful, especially if multiple genes within the same pathway are simultaneously affected.

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Morphological and molecular markers are correlated with maturation-competence of human oocytes

Abstract: We suggest that the central location of GV within the oocytes, which is associated with an absence of Fyn at the GV and the presence of thick filamentous MTs in the ooplasm, may serve as a predictor of successful maturation and provide new insights for the use of IVM.

Cited by 33 publications

References 55 publications

Regulation of GVBD in mouse oocytes by miR-125a-3p and Fyn kinase through modulation of actin filaments

Regulation of GVBD in mouse oocytes by miR-125a-3p and Fyn kinase through modulation of actin filaments

Interleukin-1 deficiency prolongs ovarian lifespan in mice

Digital multiplexed mRNA analysis of functionally important genes in single human oocytes and correlation of changes in transcript levels with oocyte protein expression

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