Time-lapse observation and transcriptome analysis of a case with repeated multiple pronuclei after IVF/ICSI

Dai, Jing; Leng, Lizhi; Lu, Can‐Zhong; Gong, Fei; Zhang, S. P.; Zheng, Wei; Lu, Guangxiu; Lin, Ge

doi:10.1007/s10815-017-0972-9

Cited by 9 publications

(6 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The MPN therefore can cause infertility and recurrent failure IVF/ICSI. In 2017, transcription levels of several genes in oocytes have been identified to alter by analysing differentially expressed gene profiles between normal and MPN zygotes 5. However, no genes have been reported to cause infertility characterised by MPN.…”

Section: Introductionmentioning

confidence: 99%

Homozygous mutations in REC114 cause female infertility characterised by multiple pronuclei formation and early embryonic arrest

et al. 2019

View full text Add to dashboard Cite

BackgroundAbnormal pronuclear formation during fertilisation and subsequent early embryonic arrest results in female infertility. In recent years, with the prevalence of assisted reproductive technology, a few genes have been identified that are involved in female infertility caused by abnormalities in oocyte development, fertilisation and embryonic development. However, the genetic factors responsible for multiple pronuclei formation during fertilisation and early embryonic arrest remain largely unknown.ObjectiveWe aim to identify genetic factors responsible for multiple pronuclei formation during fertilisation or early embryonic arrest.MethodsWhole-exome sequencing was performed in a cohort of 580 patients with abnormal fertilisation and early embryonic arrest. Effects of mutations were investigated in HEK293T cells by western blotting and immunoprecipitation, as well as minigene assay.ResultsWe identified a novel homozygous missense mutation (c.397T>G, p.C133G) and a novel homozygous donor splice-site mutation (c.546+5G>A) in the meiotic gene REC114. REC114 is involved in the formation of double strand breaks (DSBs), which initiate homologous chromosome recombination. We demonstrated that the splice-site mutation affected the normal alternative splicing of REC114, while the missense mutation reduced the protein level of REC114 in vitro and resulted in the loss of its function to protect its partner protein MEI4 from degradation.ConclusionsOur study has identified mutations in REC114 responsible for human multiple pronuclei formation and early embryonic arrest, and these findings expand our knowledge of genetic factors that are responsible for normal human female meiosis and fertility.

show abstract

Section: Introductionmentioning

confidence: 99%

Homozygous mutations in REC114 cause female infertility characterised by multiple pronuclei formation and early embryonic arrest

et al. 2019

View full text Add to dashboard Cite

show abstract

“…Furthermore, as learned from rat OSA, certain OSA-oocytes could show minimal morphological signs (e.g., sister chromatids separated or even scattered in cytoplasm but without 2nd PB or pronucleus formation). Therefore, we propose OSA should be considered for those unexplained abnormal fertilization with repeated triploid pronuclei (3PN) ( Grigoryan et al, 2019 ) or even more pronuclei (e.g., up to 8PN) ( Dai et al, 2017 ) after ICSI. Other lessons we can learn from animal models and issues that should be addressed are as follows: (1) time interval between oocyte pickup and IVF/ICSI.…”

Section: Discussionmentioning

confidence: 99%

“…Instead, OSA-oocytes enter a so-called “metaphase III-like” (M-III) arrest ( Zernicka-Goetz, 1991 ). Notably, these OSA-oocytes can be re-activated by sperm or chemicals, and once re-activated, those scattered chromatids will form multiple pronuclei (MPN), which may recapitulate certain MPN and aneuploidy cases observed in human fertility clinics ( Van Blerkom et al, 1984 ; Dozortsev et al, 1998 ; Hayes et al, 2001 ; Dai et al, 2017 ; Grigoryan et al, 2019 ). Different from rat, some hamster and human OSA-oocytes can reach interphase with visible pronuclei ( Longo, 1974 ; Van Blerkom et al, 1994 ; Sun et al, 2002 ; Jiang et al, 2015 ; Osman et al, 2019 ; Ye et al, 2020 ).…”

Section: First Type Of Osamentioning

confidence: 99%

Oocyte Spontaneous Activation: An Overlooked Cellular Event That Impairs Female Fertility in Mammals

Cui

2021

Front. Cell Dev. Biol.

View full text Add to dashboard Cite

In mammals, including humans, mature oocytes are ovulated into the oviduct for fertilization. Normally, these oocytes are arrested at metaphase of the second meiosis (MII), and this arrest can be maintained for a certain period, which is essential for fertilization in vivo and oocyte manipulations in vitro, such as assisted reproduction in clinics and nuclear/spindle transfer in laboratories. However, in some species and under certain circumstances, exit from MII occurs spontaneously without any obvious stimulation or morphological signs, which is so-called oocyte spontaneous activation (OSA). This mini-review summarizes two types of OSA. In the first type (e.g., most rat strains), oocytes can maintain MII arrest in vivo, but once removed out, oocytes undergo OSA with sister chromatids separated and eventually scattered in the cytoplasm. Because the stimulation is minimal (oocyte collection itself), this OSA is incomplete and cannot force oocytes into interphase. Notably, once re-activated by sperm or chemicals, those scattered chromatids will form multiple pronuclei (MPN), which may recapitulate certain MPN and aneuploidy cases observed in fertility clinics. The second type of OSA occurs in ovarian oocytes (e.g., certain mouse strains and dromedary camel). Without ovulation or fertilization, these OSA-oocytes can initiate intrafollicular development, but these parthenotes cannot develop to term due to aberrant genomic imprinting. Instead, they either degrade or give rise to ovarian teratomas, which have also been reported in female patients. Last but not the least, genetic models displaying OSA phenotypes and the lessons we can learn from animal OSA for human reproduction are also discussed.

show abstract

“…2) Failure of the second polar body (PBII) extrusion. 3) Abnormal pronucleus formation (17,20). In the Hong study(1), the authors speculated that the high MPN rate is due to the damage on oocytes, when the cumulus cells are completely removed at 2 ~ 4 h post-insemination.…”

Section: Discussionmentioning

confidence: 99%

“…In addition, the damage of the cytoskeleton organization also affects the pronucleus formation. And assembly error of pronucleus can result in MPN too (20). Therefore, abnormal pronucleus formation is another main cause of the highest MPN rate in the time ≤ 4 group.…”

Section: Discussionmentioning

confidence: 99%

Different Timings of Early Cumulus Cells Removal Have Different Multiple Pronuclei Rates in Human in Vitro Fertilization

Liu

Jiang

et al. 2021

Preprint

View full text Add to dashboard Cite

Background: Cumulus cells removal 4 h post-insemination has a significantly higher multiple pronuclei (MPN) rate than cumulus cells removal 20 h post-insemination. And, cumulus cells removal 6 h post-insemination has a significantly lower MPN rate than cumulus cells removal 20 h post-insemination. However, it remains unclear whether the different timings of early cumulus cells removal, such as the timings of 4, 5 and 6 h post-insemination, have significantly different MPN rates.Methods: This was a retrospective study. The included cycles were early cumulus cells removal cycles (n=752) at our center from January 2015 to August 2020. The included cycles were divided into two groups according to whether MPN exist (MPN=0% and MPN>0%). The patient and cycle stimulation characteristics of the two groups were compared. Binary logistic regression was performed to investigate the correlation between the timing of early cumulus cells removal and MPN. The cohort study was also performed to compare the patient characteristics, cycle stimulation characteristics, fertilization outcomes, and cultivation outcomes.Results: In the population of our study, the timing of early cumulus cells removal had a significant effect on the MPN. The cumulus cells removal ≤4 h post-insemination group had a high MPN rate, and the 5.5<time≤6 h group had a high fertilization failure rate. However, 2PN rate was not significantly different among the different timings of early cumulus cells removal. In addition, the ≤4 h post-insemination group had a high grade 1–2 embryo rate at day 3.Conclusion(s): Even if all the timings of cumulus cells removal are early, the different timings of early cumulus cells removal still have a significant effect on the MPN.

show abstract

Time-lapse observation and transcriptome analysis of a case with repeated multiple pronuclei after IVF/ICSI

Abstract: We identified several candidate genes affecting pronucleus formation as a new cause of infertility.

Cited by 9 publications

References 26 publications

Homozygous mutations in REC114 cause female infertility characterised by multiple pronuclei formation and early embryonic arrest

Homozygous mutations in REC114 cause female infertility characterised by multiple pronuclei formation and early embryonic arrest

Oocyte Spontaneous Activation: An Overlooked Cellular Event That Impairs Female Fertility in Mammals

Different Timings of Early Cumulus Cells Removal Have Different Multiple Pronuclei Rates in Human in Vitro Fertilization

Contact Info

Product

Resources

About