Failure of oocyte activation,including polyspermy and defects in pronuclear(PN)formation,triggers early embryonic developmental arrest.Many studies have shown that phospholipase C zeta 1(PLCZ1)mutations cause failure ...Failure of oocyte activation,including polyspermy and defects in pronuclear(PN)formation,triggers early embryonic developmental arrest.Many studies have shown that phospholipase C zeta 1(PLCZ1)mutations cause failure of PN formation following intracytoplasmic sperm injection(ICSI);however,whether PLCZ1 mutation is associated with polyspermy during in vitro fertilization(IVF)remains unknown.Whole-exome sequencing(WES)was performed to identify candidate mutations in couples with primary infertility.Sanger sequencing was used to validate the mutations.Multiple PLCZ1-mutated sperm were injected into human and mouse oocytes to explore whether PN formation was induced.Assisted oocyte activation(AOA)after ICSI was performed to overcome the failure of oocyte activation.We identified three PLCZ1 mutations in three patients who experienced polyspermy during IVF cycles,including a novel missense mutation c.1154C>T,p.R385Q.PN formation failure was observed during the ICSI cycle.However,injection of multiple PLCZ1-mutated sperm induced PN formation,suggesting that the Ca2+oscillations induced by the sperm exceeded the necessary threshold for PN formation.AOA after ICSI enabled normal fertilization,and all patients achieved successful pregnancies.These findings expand the mutational spectrum of PLCZ1 and suggest an important role for PLCZ1 in terms of blocking polyspermy.Furthermore,this study may benefit genetic diagnoses in cases of abnormal fertilization and provide potential appropriate therapeutic measures for these patients with sperm-derived polyspermy.展开更多
目的:研究卵胞浆内精子注射(ICSI)周期卵母细胞滑面内质网(sER)聚集对早期胚胎体外发育及妊娠结局的影响并分析其原因。方法:99例患者ICSI治疗107个周期,于HCG日测定血清激素E2、P、LH浓度以及子宫内膜厚度。根据所获MⅡ期卵母细胞是否...目的:研究卵胞浆内精子注射(ICSI)周期卵母细胞滑面内质网(sER)聚集对早期胚胎体外发育及妊娠结局的影响并分析其原因。方法:99例患者ICSI治疗107个周期,于HCG日测定血清激素E2、P、LH浓度以及子宫内膜厚度。根据所获MⅡ期卵母细胞是否出现sER聚集分为A、B两组,A组为所获MⅡ期卵母细胞均未出现sER聚集周期共90例;B组为有至少一枚MⅡ期卵母细胞出现sER聚集周期,共17例。比较A、B两组HCG日E2、P、LH水平、子宫内膜厚度,及治疗周期数、年龄、病程、Gn总量、Gn天数、获卵数、受精率、可用胚胎率、优质胚胎率、移植胚胎数、周期临床妊娠率、种植率、流产率和原发、继发不孕患者比例的差异。并且对比B组sER聚集阳性[sER(+)]与sER聚集阴性[sER(-)]卵母细胞的受精率、可用胚胎率和优质胚胎率。结果:B组HCG日E2水平明显高于A组(P<0.05,3141.18±604.47 vs 2635.12±825.46),而两组间P和LH水平以及子宫内膜厚度均无显著差异(P>0.05)。B组优质胚胎率显著低于A组(P<0.05,47.83%vs 57.67%),而B组Gn天数(P<0.01,13.35±1.66 vs 11.83±2.4)和流产率(P<0.05,100%vs 17.86%)均明显高于A组,两组在年龄、治疗周期数、病程、Gn总量、获卵数、受精率、可用胚胎率、移植胚胎数、临床妊娠率、种植率以及原发、继发不孕患者比例等各方面均无显著差异(P>0.05)。B组sER(+)卵母细胞的优质胚胎率显著低于sER-卵母细胞(P<0.01,20.69%vs 52.9%),而受精率和可用胚胎率无显著性差异(P>0.05)。结论:ICSI周期卵母细胞sER聚集对早期胚胎体外发育及妊娠结局均有不良影响,sER聚集可能与HCG日高E2水平及长时间Gn刺激有关。展开更多
基金the General Project of Chongqing Natural Science foundation of China(cstc2021jcyj-msxmX0877)the Chongqing Medical Scientific Research Project(Joint Project of Chongqing Health Commission and Science and Technology Bureau,2023MSXM054)the General Project of Chongqing Health Center for Women and Children(2020YJMS01 and 2021YJMS05).
文摘Failure of oocyte activation,including polyspermy and defects in pronuclear(PN)formation,triggers early embryonic developmental arrest.Many studies have shown that phospholipase C zeta 1(PLCZ1)mutations cause failure of PN formation following intracytoplasmic sperm injection(ICSI);however,whether PLCZ1 mutation is associated with polyspermy during in vitro fertilization(IVF)remains unknown.Whole-exome sequencing(WES)was performed to identify candidate mutations in couples with primary infertility.Sanger sequencing was used to validate the mutations.Multiple PLCZ1-mutated sperm were injected into human and mouse oocytes to explore whether PN formation was induced.Assisted oocyte activation(AOA)after ICSI was performed to overcome the failure of oocyte activation.We identified three PLCZ1 mutations in three patients who experienced polyspermy during IVF cycles,including a novel missense mutation c.1154C>T,p.R385Q.PN formation failure was observed during the ICSI cycle.However,injection of multiple PLCZ1-mutated sperm induced PN formation,suggesting that the Ca2+oscillations induced by the sperm exceeded the necessary threshold for PN formation.AOA after ICSI enabled normal fertilization,and all patients achieved successful pregnancies.These findings expand the mutational spectrum of PLCZ1 and suggest an important role for PLCZ1 in terms of blocking polyspermy.Furthermore,this study may benefit genetic diagnoses in cases of abnormal fertilization and provide potential appropriate therapeutic measures for these patients with sperm-derived polyspermy.
文摘目的:研究卵胞浆内精子注射(ICSI)周期卵母细胞滑面内质网(sER)聚集对早期胚胎体外发育及妊娠结局的影响并分析其原因。方法:99例患者ICSI治疗107个周期,于HCG日测定血清激素E2、P、LH浓度以及子宫内膜厚度。根据所获MⅡ期卵母细胞是否出现sER聚集分为A、B两组,A组为所获MⅡ期卵母细胞均未出现sER聚集周期共90例;B组为有至少一枚MⅡ期卵母细胞出现sER聚集周期,共17例。比较A、B两组HCG日E2、P、LH水平、子宫内膜厚度,及治疗周期数、年龄、病程、Gn总量、Gn天数、获卵数、受精率、可用胚胎率、优质胚胎率、移植胚胎数、周期临床妊娠率、种植率、流产率和原发、继发不孕患者比例的差异。并且对比B组sER聚集阳性[sER(+)]与sER聚集阴性[sER(-)]卵母细胞的受精率、可用胚胎率和优质胚胎率。结果:B组HCG日E2水平明显高于A组(P<0.05,3141.18±604.47 vs 2635.12±825.46),而两组间P和LH水平以及子宫内膜厚度均无显著差异(P>0.05)。B组优质胚胎率显著低于A组(P<0.05,47.83%vs 57.67%),而B组Gn天数(P<0.01,13.35±1.66 vs 11.83±2.4)和流产率(P<0.05,100%vs 17.86%)均明显高于A组,两组在年龄、治疗周期数、病程、Gn总量、获卵数、受精率、可用胚胎率、移植胚胎数、临床妊娠率、种植率以及原发、继发不孕患者比例等各方面均无显著差异(P>0.05)。B组sER(+)卵母细胞的优质胚胎率显著低于sER-卵母细胞(P<0.01,20.69%vs 52.9%),而受精率和可用胚胎率无显著性差异(P>0.05)。结论:ICSI周期卵母细胞sER聚集对早期胚胎体外发育及妊娠结局均有不良影响,sER聚集可能与HCG日高E2水平及长时间Gn刺激有关。
