In Vitro Maturation of Mouse Oocytes Increases the Level of Kif11/Eg5 on Meiosis II Spindles

Kovacovicova, Kristina; Awadová, Thuraya; Mikel, Pavel; Anger, Martin

doi:10.1095/biolreprod.115.133900

Cited by 16 publications

(14 citation statements)

References 37 publications

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“…Microinjection was done using an IM-300 microinjector (Narishige) on a Leica DM IL inverted microscope as described previously [Kovacovicova et al, 2016]. cRNAs for microinjection were prepared by in vitro transcription (mMESSAGE mMACHINE T3 in vitro transcription kit, Life Technologies) of plasmids containing ORFs of mouse securin, β-tubulin, and histone 2B (H2B) in the transcription fusion with fluorescent proteins CFP, Venus, and mCherry, respectively.…”

Section: Microinjectionmentioning

confidence: 99%

See 1 more Smart Citation

True Nondisjunction of Whole Bivalents in Oocytes with Attachment and Congression Defects

Šodek

Kovacovicova

Anger

2017

Cytogenet Genome Res

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Chromosome segregation in mammalian oocytes is prone to errors causing aneuploidy with consequences such as precocious termination of development or severe developmental disorders. Aneuploidy also represents a serious problem in procedures utilizing mammalian gametes and early embryos in vitro. In our study, we focused on congression defects during meiosis I and observed whole nondisjoined bivalents in meiosis II as a direct consequence, together with a substantially delayed first polar body extrusion. We also show that the congression defects are accompanied by less stable attachments of the kinetochores. Our results describe a process by which congression defects directly contribute to aneuploidy.

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Section: Microinjectionmentioning

confidence: 99%

“…If not stated otherwise, all chemicals were derived from Sigma Aldrich, and protocols from Lane et al [2012], Kovacovicova et al [2016], and Chmátal et al [2015] were adopted. Briefly, the oocytes were incubated in pre-cooled M2 medium (4-8 ° C) for 7 min before fixation with 2% paraformaldehyde.…”

Section: Cold-stable Microtubulesmentioning

confidence: 99%

True Nondisjunction of Whole Bivalents in Oocytes with Attachment and Congression Defects

Šodek

Kovacovicova

Anger

2017

Cytogenet Genome Res

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“…Eg5 inhibition leads to the collapse of meiosis II spindles and chromosome missegregation in mammalian oocytes [30]. Eg5 is expressed in all developmental stages during porcine oocyte meiosis, and its inhibition strongly delays meiotic progression due to the reduction of Cdc2 phosphorylation with its dose-dependent property [53].…”

Section: Discussionmentioning

confidence: 99%

“…In mammalian oocytes, Eg5 regulates meiotic spindle assembly [26][27][28] and poleward tubulin flux [29]. Eg5 is required for the maintenance of bipolar spindle in mouse oocytes during meiosis II [30]. However, roles and mechanisms of Eg5 in male meiotic division remain largely unknown.…”

Section: Introductionmentioning

confidence: 99%

Kinesin-5 Eg5 is essential for spindle assembly and chromosome alignment of mouse spermatocytes

She

Zhong

et al. 2020

Cell Div

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Background: Microtubule organization is essential for bipolar spindle assembly and chromosome segregation, which contribute to genome stability. Kinesin-5 Eg5 is known to be a crucial regulator in centrosome separation and spindle assembly in mammalian somatic cells, however, the functions and mechanisms of Eg5 in male meiotic cell division remain largely unknown. Results: In this study, we have found that Eg5 proteins are expressed in mouse spermatogonia, spermatocytes and spermatids. After Eg5 inhibition by specific inhibitors Monastrol, STLC and Dimethylenastron, the meiotic spindles of dividing spermatocytes show spindle collapse and the defects in bipolar spindle formation. We demonstrate that Eg5 regulates spindle bipolarity and the maintenance of meiotic spindles in meiosis. Eg5 inhibition leads to monopolar spindles, spindle abnormalities and chromosome misalignment in cultured GC-2 spd cells. Furthermore, Eg5 inhibition results in the decrease of the spermatids and the abnormalities in mature sperms. Conclusions: Our results have revealed an important role of kinesin-5 Eg5 in male meiosis and the maintenance of male fertility. We demonstrate that Eg5 is crucial for bipolar spindle assembly and chromosome alignment in dividing spermatocytes. Our data provide insights into the functions of Eg5 in meiotic spindle assembly of dividing spermatocytes.

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“…Although low concentrations of hCG led to the maturation of the oocytes, the most effective condition for the maturation of such non-induced oocytes was a combination of hCG (100 IU/L) with EGF (5 mg/mL) in Universal IVF medium. The concentration ranges of these hormones were relatively higher than those used in mice [42,43]. Some studies have revealed that the concentrations used for marmosets in pre-gonadotropin-induced IVM ranged from 1 to 10 IU with or without a combination of other factors, such as FSH and estradiol [38,44,45,46].…”

Section: Discussionmentioning

confidence: 99%

Expression of Transcripts in Marmoset Oocytes Retrieved during Follicle Isolation Without Gonadotropin Induction

Kim

Kang

Yun

et al. 2019

IJMS

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The in vitro maturation of oocytes is frequently used as an assisted reproductive technique (ART), and has been successfully established in humans and rodents. To overcome the limitations of ART, novel procedures for the in vitro maturation of early follicles are emerging. During the follicle isolation procedure, the unintended rupture of each follicle leads to a release of extra oocytes. Such oocytes, which are obtained during follicle isolation from marmosets, can be used for early maturation studies. Marmoset (Callithrix jacchus), which is classified as a new-world monkey, is a novel model that has been employed in reproductive biomedical research, as its reproductive physiology is similar to that of humans in several aspects. The ovaries of female marmosets were collected, and the excess oocytes present during follicle isolation were retrieved without pre-gonadotropin induction. Each oocyte was matured in vitro for 48 h in the presence of various concentrations of human chorionic gonadotropin (hCG) and epidermal growth factor (EGF), and the maturity of oocytes and optimal maturation conditions were evaluated. Each oocyte was individually reverse-transcribed, and the expression of mRNAs and microRNAs (miRs) were analyzed. Concentrations of hCG significantly affected the maturation rate of oocytes [the number of metaphase II (MII) oocytes]. The expression of BMP15 and ZP1 was highest when the oocytes were matured using 100 IU/L of hCG without pre-treatment with gonadotropins, and that of Cja-mir-27a was highest when cultured with follicle stimulating hormone. These results suggest that these up-regulated miRs affect the maturation of oocytes. Interactions with other protein networks were analyzed, and a strong association of BMP15 and ZP1 with sperm binding receptor (ACR), anti-Müllerian hormone (AMH), and AMH receptor was demonstrated, which is related to the proliferation of granulosa cells. Collectively, on the basis of these results, the authors propose optimal maturation conditions of excess oocytes of marmoset without in vivo gonadotropin treatment, and demonstrated the roles of miRs in early oocyte maturation at the single-cell level in marmosets.

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In Vitro Maturation of Mouse Oocytes Increases the Level of Kif11/Eg5 on Meiosis II Spindles

Cited by 16 publications

References 37 publications

True Nondisjunction of Whole Bivalents in Oocytes with Attachment and Congression Defects

True Nondisjunction of Whole Bivalents in Oocytes with Attachment and Congression Defects

Kinesin-5 Eg5 is essential for spindle assembly and chromosome alignment of mouse spermatocytes

Expression of Transcripts in Marmoset Oocytes Retrieved during Follicle Isolation Without Gonadotropin Induction

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