scholarly journals DNA methylation and gene expression changes in mouse pre- and post-implantation embryos generated by intracytoplasmic sperm injection with artificial oocyte activation

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Mingru Yin ◽  
Weina Yu ◽  
Wenzhi Li ◽  
Qianqian Zhu ◽  
Hui Long ◽  
...  
Keyword(s):  
Gene Expression ◽  
Birth Weight ◽  
Intracytoplasmic Sperm Injection ◽  
Oocyte Activation ◽  
Blastocyst Formation ◽  
Developmental Potential ◽  
Imprinted Gene ◽  
Reproduction Biology ◽  
Artificial Oocyte Activation ◽  
Post Implantation

Abstract Background The application of artificial oocyte activation (AOA) after intracytoplasmic sperm injection (ICSI) is successful in mitigating fertilization failure problems in assisted reproductive technology (ART). Nevertheless, there is no relevant study to investigate whether AOA procedures increase developmental risk by disturbing subsequent gene expression at different embryonic development stages. Methods We used a mouse model to explore the influence of AOA treatment on pre- and post-implantation events. Firstly, the developmental potential of embryos with or without AOA treatment were assessed by the rates of fertilization and blastocyst formation. Secondly, transcriptome high-throughput sequencing was performed among the three groups (ICSI, ICSI-AOA and dICSI-AOA groups). The hierarchical clustering and Principal Component Analysis (PCA) analysis were used. Subsequently, Igf2r/Airn methylation analysis were detected using methylation-specific PCR sequencing following bisulfite treatment. Finally, birth rate and birth weight were examined following mouse embryo transfer. Results The rates of fertilization and blastocyst formation were significantly lower in oocyte activation-deficient sperm injection group (dICSI group) when compared with the ICSI group (30.8 % vs. 84.4 %, 10.0 % vs. 41.5 %). There were 133 differentially expressed genes (DEGs) between the ICSI-AOA group and ICSI group, and 266 DEGs between the dICSI-AOA group and ICSI group. In addition, the imprinted gene, Igf2r is up regulated in AOA treatment group compared to control group. The Igf2r/Airn imprinted expression model demonstrates that AOA treatment stimulates maternal allele-specific mehtylation spreads at differentially methylated region 2, followed by the initiation of paternal imprinted Airn long non-coding (lnc) RNA, resulting in the up regulated expression of Igf2r. Furthermore, the birth weight of newborn mice originating from AOA group was significantly lower compared to that of ICSI group. The pups born following AOA treatment did not show any other abnormalities during early development. All offspring mated successfully with fertile controls. Conclusions AOA treatment affects imprinted gene Igf2r expression and mehtylation states in mouse pre- and post-implantation embryo, which is regulated by the imprinted Airn. Nevertheless, no significant differences were found in post-natal growth of the pups in the present study. It is hoped that this study could provide valuable insights of AOA technology in assisted reproduction biology.

scholarly journals Risk of birth defects in children conceived by artificial oocyte activation and intracytoplasmic sperm injection: a meta-analysis

2020 ◽  
Vol 18 (1) ◽  
Author(s):  
Rui Long ◽  
Meng Wang ◽  
Qi Yu Yang ◽  
Shi Qiao Hu ◽  
Li Xia Zhu ◽  
...  
Keyword(s):  
Birth Defects ◽  
Chromosomal Aberrations ◽  
Intracytoplasmic Sperm Injection ◽  
Safety Concern ◽  
Final Analysis ◽  
Oocyte Activation ◽  
Patient Counseling ◽  
China National Knowledge Infrastructure ◽  
Significant Difference ◽  
Artificial Oocyte Activation

Abstract Background Whether artificial oocyte activation (ICSI-AOA) will increase the risk of birth defects remains controversial. Thus, we performed this study to evaluate the risk of birth defects and further compare the incidence of different birth defects types (chromosomal aberrations and non-chromosomal aberrations) in children conceived by ICSI-AOA and conventional intracytoplasmic sperm injection (ICSI) in an enlarged sample size. Method A comprehensive review of the literatures comparing birth defects in children conceived by ICSI-AOA and conventional ICSI by October 2020 was performed in PubMed, Embase, Cochrane Libraries, Web of Science, and Chinese databases including China National Knowledge Infrastructure, China Biology Medicine disc and Wan Fang. Risk ratios (RR) and 95% confidence intervals (CI) were calculated. Results Five studies were included in the final analysis. Compared with conventional ICSI, ICSI-AOA did not increase the birth defects rate (RR = 1.27, 95%CI 0.70–2.28) of children. Furthermore, in a subgroup analysis, birth defects were classified into two types (chromosomal aberrations and non-chromosomal aberrations) in four studies and no statistical difference were revealed. Conclusion Our analysis indicates that ICSI-AOA represents no significant difference in the prevalence of major birth defects or types of birth defects (chromosomal aberrations and non-chromosomal aberrations) comparing with conventional ICSI. This conclusion may provide clinicians evidence-based support in patient counseling and instruction of the application and safety concern about ICSI-AOA.

scholarly journals The employment of strontium to activate mouse oocytes: effects on spermatid-injection outcome

Reproduction ◽  
2006 ◽  
Vol 131 (2) ◽  
pp. 259-267 ◽  
Author(s):  
Jean Loren ◽  
Orly Lacham-Kaplan
Keyword(s):  
Birth Outcomes ◽  
Round Spermatid ◽  
Oocyte Activation ◽  
Embryo Viability ◽  
Blastocyst Development ◽  
Incubation Periods ◽  
Artificial Oocyte Activation ◽  
The Impact ◽  
Post Implantation ◽  
Round Spermatid Injection

The present research investigated the effects of various strontium concentrations, in combination with different incubation periods, on mouse parthenogentic oocyte activation and blastocyst development. The results for blastocyst development showed a trend indicating that 10 mM strontium for 3 h was the optimal strontium protocol. Ethanol, an agent that incites oocyte activation via a monotonic rise in calcium, was employed as a control. The outcome of blastocyst formation arising from parthenogenic ethanol activation was significantly less (P < 0.001) than that achieved by the optimal strontium protocol. To assess the impact of strontium oocyte activation on embryo viability following fertilization with immature germ cells, the protocol of 10 mM strontium for 3 h was applied to oocytes injected with round spermatids and then compared with other protocols. The results indicate that following round-spermatid injection the benefits derived from strontium artificial oocyte activation are evident during both pre- and post-implantation development. However, in order to adjust the protocol to the most effective round-spermatid injection in relation to the oocyte cell cycle, injection was done 1.5 h after strontium activation followed by another 1.5 h activation in strontium. The implementation of round-spermatid injection in combination with this oocyte-activation protocol led to live-birth outcomes not significantly different to those outcomes obtained by mature spermatozoa.

151 A NOVEL METHOD TO INCREASE THE DEVELOPMENTAL POTENTIAL OF ACTIVATED OOCYTES BY USING THE Zn2+ CHELATOR TPEN [N,N,N',N'-TETRAKIS(2-PYRIDYLMETHYL)ETHYLENEDIAMINE]

2014 ◽  
Vol 26 (1) ◽  
pp. 189
Author(s):  
K. Lee ◽  
A. Davis ◽  
C. N. Murphy ◽  
R. S. Prather
Keyword(s):  
Positive Feedback ◽  
Feedback Loop ◽  
Cell Number ◽  
Total Cell ◽  
Control Group ◽  
Oocyte Activation ◽  
Blastocyst Formation ◽  
High Concentration ◽  
Positive Feedback Loop ◽  
Developmental Potential

An increase in intracellular Ca2+ concentration is essential for oocyte activation. Thus most artificial oocyte-activation methods focus on increasing the Ca2+ concentration in the oocytes. Recently, full-term development was reported in mice when oocytes were activated with no increase in intracellular Ca2+ using the Zn2+ chelator N,N,N′,N′-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN). During oocyte maturation, Zn2+ is responsible for Cdc25c recognition. Once attached, the Cdc25c-zinc complex dephosphorylates maturation-promoting factor (MPF)/cyclin-dependent kinase 1 (cdk1), thus activating the Cdc25c-MPF positive feedback loop and keeping the oocyte suspended in metaphase II. The TPEN can inhibit the Cdc25c-MPF positive feedback loop, indirectly giving TPEN the power to degrade MPF, allowing the oocyte to exit metaphase II. First, we tested if incubation of porcine oocytes with TPEN could induce oocyte activation. Second, we examined whether the combination of TPEN with conventional activation methods could increase the developmental potential of activated oocytes. Last, based on the results, somatic cell nuclear transfer (SCNT) embryos were further activated with the optimum condition of TPEN to produce clones to confirm developmental competence. Frequencies of blastocyst formation were recorded and analysed by using ANOVA following arcsin transformation. Total cell numbers in blastocysts were counted and compared by using the Student's t-test. Differences at P < 0.05 were considered significant. When oocytes were incubated with a high concentration of TPEN (100–250 μM) for 10 to 120 min, blastocyst formation was comparable with conventional activation methods; however, the total cell number in the blastocysts was significantly lower (31.3 ± 3.1 v. 24.8 ± 1.9). When oocytes were activated with conventional methods and then incubated with the high concentration of TPEN, embryo development was drastically decreased; no blastocyst development was achieved from TPEN-treated oocytes. Interestingly, when activated oocytes were incubated with a low concentration of TPEN (5–10 μM), surprisingly, the TPEN-treated group showed higher developmental potential compared with the control group. Specifically, the average percent blastocyst formation of TPEN-treated oocytes (5 μM for 30 min) was 27.2 ± 1.7%, but only 10.6 ± 2.5% developed to blastocyst in the control group. Moreover, the average cell number in blastocysts was significantly higher in TPEN-treated oocytes compared with the control group (33.1 ± 2.6 v. 28.2 ± 2.1, respectively). When 290 chemically activated SCNT embryos were treated with 5 μM TPEN for 30 min and transferred into a surrogate, 2 healthy piglets were born. The results indicate that incubation of oocytes with TPEN alone can activate porcine oocytes. Also, when activated oocytes are incubated with the right concentration of TPEN, it can increase embryo quality in vitro. Embryo transfer results also show that TPEN-incubated SCNT embryos are developmentally competent. Additional studies would guide us to develop more efficient way to use TPEN in the activation of SCNT embryos.

Artificial oocyte activation with calcium ionophore A23187 in intracytoplasmic sperm injection cycles using surgically retrieved spermatozoa

2009 ◽  
Vol 92 (1) ◽  
pp. 131-136 ◽  
Author(s):  
Edson Borges Jr. ◽  
Daniela Paes de Almeida Ferreira Braga ◽  
Tatiana Carvalho de Sousa Bonetti ◽  
Assumpto Iaconelli Jr. ◽  
José Gonçalves Franco Jr.
Keyword(s):  
Intracytoplasmic Sperm Injection ◽  
Calcium Ionophore ◽  
Calcium Ionophore A23187 ◽  
Ionophore A23187 ◽  
Oocyte Activation ◽  
Artificial Oocyte Activation

Reproductive Outcomes of Artificial Oocyte Activation after ICSI Undergoing Blastocyst Transfer: A 6-year Retrospective Cohort Study

2021 ◽  
Author(s):  
Xuehua Yan ◽  
Xitong Liu ◽  
Wennan Chen ◽  
Xueli Yan ◽  
Yating Zhu ◽  
...  
Keyword(s):  
Logistic Regression ◽  
Regression Analysis ◽  
Pregnancy Rate ◽  
Logistic Regression Analysis ◽  
Intracytoplasmic Sperm Injection ◽  
Clinical Pregnancy ◽  
Blastocyst Transfer ◽  
Multivariable Logistic Regression Analysis ◽  
Oocyte Activation ◽  
Artificial Oocyte Activation

Abstract Research question: Is the pregnancy rate affected by artificial oocyte activation (AOA) with A23187 after intracytoplasmic sperm injection (ICSI) in infertile patients?Design: Our retrospective study included 308 patients who transferred blastocyst from routine intracytoplasmic sperm injection (ICSI) and 82 patients who transferred blastocyst from routine ICSI combined with AOA (ICSI-AOA) from January 2014 to April 2020. Pregnancy outcomes of couples who received routine ICSI or ICSI-AOA were analyzed after the first blastocyst transfer, which covered frozen-thawed blastocyst transfer and fresh blastocyst transfer. AOA was performed with A237817. We used multivariable logistic regression analysis to determine which variables could be independently associated with the pregnancy rate. Effect sizes were summarized as odds ratios (ORs), with precision evaluated by 95% CIs. Results: The clinical pregnancy rate was 71.95% in the AOA group and 57.47% in the routine ICSI group. The effect size of the AOA on clinical rate was evaluated in prespecified and exploratory subgroups in each subgroup. And multivariable logistic regression analysis was performed to identify factors associated with the clinical rate. The AOA group had a higher chance of clinical pregnancy in all subgroups: female age at oocyte retrieval, female BMI, protocol in the fresh cycle, female infertility type, MII oocyte numbers, fresh or frozen blastocyst transfer, No. of blastocyst transfer and blastocyst quality. Multivariable analysis showed AOA to be associated with an increased likelihood of clinical pregnancy compared with routine ICSI (p=0.03; adjusted OR 1.89, 95% CI 1.09–3.27).Conclusions: This study suggested that AOA can increase the rate of clinical pregnancy obviously, which helps clinicians to advise patients on AOA risks.

Artificial oocyte activation in severe teratozoospermia undergoing intracytoplasmic sperm injection

2008 ◽  
Vol 90 (6) ◽  
pp. 2231-2237 ◽  
Author(s):  
Mohammad H. Nasr-Esfahani ◽  
Shahnaz Razavi ◽  
Zeinab Javdan ◽  
Marzeyeh Tavalaee
Keyword(s):  
Intracytoplasmic Sperm Injection ◽  
Oocyte Activation ◽  
Artificial Oocyte Activation

scholarly journals Effects of Assisted Reproduction Technology on Placental Imprinted Gene Expression

2010 ◽  
Vol 2010 ◽  
pp. 1-4 ◽  
Author(s):  
Yukiko Katagiri ◽  
Chizu Aoki ◽  
Yuko Tamaki-Ishihara ◽  
Yusuke Fukuda ◽  
Mamoru Kitamura ◽  
...  
Keyword(s):  
Gene Expression ◽  
Birth Weight ◽  
Assisted Reproduction ◽  
Expression Patterns ◽  
Placental Tissue ◽  
Gene Expression Patterns ◽  
Fetal Growth Retardation ◽  
Imprinted Gene ◽  
Assisted Reproduction Technology ◽  
Significant Difference

We used placental tissue to compare the imprinted gene expression of IGF2, H19, KCNQ1OT1, and CDKN1C of singletons conceived via assisted reproduction technology (ART) with that of spontaneously conceived (SC) singletons. Of 989 singletons examined (ART ; SC ), neonatal weight was significantly lower in the ART group than in the SC group, but placental weight showed no significant difference. Gene expression analyzed by real-time PCR was similar for both groups with appropriate-for-date (AFD) birth weight. H19 expression was suppressed in fetal growth retardation (FGR) cases in the ART and SC groups compared with AFD cases ( and , resp.). In contrast, CDKN1C expression was suppressed in FGR cases in the ART group , while KCNQ1OT1 expression was hyperexpressed in FGR cases in the SC group . As imprinted gene expression patterns differed between the ART and SC groups, we speculate that ART modifies epigenetic status even though the possibilities always exist.

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