sq-23377 and Infertility--Male

Artificial oocyte activation (AOA) is considered an effective method to improve clinical outcomes in patients with some forms of male factor infertility and does not increase the risk of birth defects. However, the effects of AOA on patients with multiple morphological abnormalities of the sperm flagella (MMAF) caused by a

Topics: Female; Flagella; Humans; Infertility, Male; Ionomycin; Male; Oocytes; Pregnancy; Pregnancy Rate; Semen; Spermatozoa

To evaluate efficiency of ionomycin on fertilization and cleavage rates, embryo development, and pregnancy rate after intracytoplasmic sperm injection (ICSI) in teratozoospermic patients.. Laboratory study.. Isfahan fertility and infertility Center, and Royan institute, Tehran, Iran.. Eighty-seven couples with male factor etiology (severe teratozoospermia) undergoing ICSI.. Ionomycin for artificial oocyte activation.. After oocyte collection, the oocytes were randomly divided into two groups: control and artificial oocyte activation (AOA). The injected oocytes in the control group were cultured in G1. The remaining oocytes were chemically activated by exposure to 10 muM ionomycin for 10 minutes. Around 16 to 18 hours after ICSI, fertilization was assessed by the presence of pronuclei. The percentage of cleavage and high-quality embryos were calculated 48 and 72 hours after ICSI. Clinical pregnancy also was determined based on ultrasound observation of a fetal heart beat.. There are significant differences in the mean of fertilization and cleavage rates 72 hours after ICSI between AOA and control groups. In patients who had no fertilization in the control group and all the embryos for transfer derived from the AOA group, two pregnancies were recorded. In the patients who had poor fertilization rate (1%-33%) in the control group (14.30%), there was a significant increase in mean fertilization rate (58.31%) because of AOA. The results of the present study also reveal that the samples had motile sperm, whereas in the chemically activated group this difference was not significant. CONCLUSION (S): It can be concluded that in cases with severe teratozoospermia AOA may improve fertilization and cleavage rates, which in turn, affect the implantation and pregnancy rate.

Topics: Cleavage Stage, Ovum; Embryo Implantation; Embryo Transfer; Female; Fertility Agents; Humans; Infertility, Male; Ionomycin; Male; Oocyte Retrieval; Oocytes; Parthenogenesis; Pregnancy; Pregnancy Rate; Severity of Illness Index; Sperm Injections, Intracytoplasmic; Treatment Outcome

Does artificial oocyte activation (AOA) by a calcium ionophore (ionomycin) improve the previous fertilization failure or poor embryo development of intracytoplasmic sperm injection (ICSI) account for male factor infertility or other infertility causes?. This retrospective study involved 114 patients receiving ICSI-AOA in Shanghai First Maternity and Infant Hospital with previous ICSI fertilization failure or poor embryo development. The previous ICSI cycles of the same patients without AOA served as the control group. The fertilization rates, cleavage rates, transferable embryo rates and blastocyst formation rates of the two groups were compared. Additionally, the clinical pregnancy, implantation rate and live birth rates were also compared to assess the efficiency and safety of AOA. Furthermore, two subgroup analyses were performed in this study based on the cause of infertility and the reason for AOA. The fertilization rate, embryonic development potential and clinical outcome were compared among groups.. Among 114 ICSI-AOA cycles, the fertilization rate, top-quality embryo rate, implantation rate, clinical pregnancy per patient and live birth rate per patient were improved significantly compared with previous ICSI cycles (p<0.05 to P< 0.001), and the miscarriage rate in the AOA group was significantly lower than that of the control group (p<0.001). In the AOA subgroups based on the cause of infertility, the fertilization rates of each subgroup were significantly improved compared with previous control cycles except for the mixed factor infertility subgroup (p<0.05 to p<0.001). In the AOA subgroups based on the reason for AOA, the fertilization rates of each subgroup were significantly increased compared with those in their previous ICSI cycle without AOA (p<0.001); however, there was no significant difference in the top-quality embryo rate. No significant improvement was found in the implantation rates and the clinical pregnancy rate in each subgroup except for the poor embryo development subgroup. In the 114 AOA cycles, 35 healthy infants (21 singletons and 7 twins) were delivered without major congenital birth defects or malformations.. This study showed that AOA with the calcium ionophore ionomycin can improve the reproductive outcomes of patients with previous fertilization failure and poor embryo development after ICSI.

Topics: Calcium Ionophores; China; Embryonic Development; Female; Fertilization; Humans; Infertility, Male; Ionomycin; Ionophores; Male; Pregnancy; Retrospective Studies; Semen; Sperm Injections, Intracytoplasmic

Fertilization involves a series of molecular events immediately following egg-sperm fusion; Ca2+ oscillations are the earliest signaling event, and they initiate the downstream reactions including pronucleus formation. Successful human reproduction requires normal fertilization. In clinical IVF or ICSI attempts, some infertile couples suffer from recurrent fertilization failure. However, the genetic reasons for fertilization failure are largely unknown. Here, we recruited several couples diagnosed with fertilization failure even though their gametes are morphologically normal. Through whole-exome sequencing and Sanger sequencing, we identified biallelic mutations in gene-encoding phospholipase C zeta 1 (PLCZ1) in four independent males in couples diagnosed with fertilization failure. Western blotting showed that missense mutations decreased the level of PLCZ1 and that nonsense or frameshift mutations resulted in undetectable or truncated proteins. Expression of these mutations in mice significantly reduced the levels of oocyte activation. Artificial oocyte activation in patient oocytes could rescue the phenotype of fertilization failure and help establish pregnancy and lead to live birth. Our findings expand the spectrum of PLCZ1 mutations that are responsible for human fertilization failure and provide a potentially feasible therapeutic treatment for these patients.

Topics: Adult; Amino Acid Sequence; Animals; Calcium Signaling; Codon, Nonsense; Conserved Sequence; Exome Sequencing; Fertilization; Fertilization in Vitro; Frameshift Mutation; Genetic Vectors; HEK293 Cells; Humans; Infertility, Male; Ionomycin; Male; Mammals; Mice; Mice, Inbred ICR; Middle Aged; Mutation, Missense; Oocytes; Pedigree; Phosphoinositide Phospholipase C; Sequence Alignment; Sequence Homology, Amino Acid; Sperm Injections, Intracytoplasmic; Treatment Failure

Oxidative stress contributes importantly to the aetiology of male infertility, impairing sperm function. The protective effect of antioxidants on seminal parameters has been established, and the antioxidant penicillamine has shown beneficial effects; however, its protective effect on human spermatozoa exposed to oxidative stress has not been reported. The objective of this work was to evaluate the effect of penicillamine on human spermatozoa exposed in vitro to oxidative stress. First, the effect of penicillamine on spermatozoa from normozoospermic donors was evaluated. Then, the effect of penicillamine on spermatozoa exposed to oxidative stress induced separately by ionomycin and hydrogen peroxide (H

Topics: Antioxidants; Culture Media; Humans; Hydrogen Peroxide; Infertility, Male; Ionomycin; Male; Membrane Potential, Mitochondrial; Oxidative Stress; Penicillamine; Reactive Oxygen Species; Reproductive Techniques, Assisted; Semen Preservation; Sperm Motility; Spermatozoa

Artificial oocyte activation (AOA) has shown to improve fertility in severe male infertility following intracytoplasmic sperm insemination (ICSI). However, the effect of AOA on the health status of children has not been studied. This pilot historical cohort study aims to evaluate physical and mental health of 79 and 89 children from 275 and 406 couples undergoing ICSI-AOA using ionomycin and conventional ICSI, respectively. The outcomes assessed were clinical pregnancy, abortion, type of delivery, and health of children (major birth defect, mental and behavior status). No significant differences were observed between the ICSI-AOA and the ICSI groups for these parameters, and the rate of major birth defects were not significantly different between the 2 groups. In this study, AOA has not imposed a greater risk on physical and mental health of children born through AOA, but for such a solid conclusion, further trails with higher number of cases are required and conclusions drawn are limited to this study.

Topics: Age Factors; Calcium Ionophores; Calcium Signaling; Child Behavior; Child Development; Child, Preschool; Female; Health Status; Humans; Infant; Infertility, Male; Ionomycin; Male; Mental Health; Oocytes; Pilot Projects; Pregnancy; Pregnancy Complications; Risk Factors; Sperm Injections, Intracytoplasmic; Treatment Outcome

Artificial oocyte activation to overcome failed fertilization after intracytoplasmic sperm injection (ICSI) in human oocytes typically employs Ca(2+) ionophores to produce a single cytosolic Ca(2+) increase. In contrast, recombinant phospholipase Czeta (PLCζ) causes Ca(2+) oscillations indistinguishable from those occurring during fertilization, but remains untested for its efficacy in a scenario of ICSI fertilization failure. Here, we compare PLCζ with other activation stimuli in a mouse model of failed oocyte activation after ICSI, in which heat-treated sperm are injected into mouse oocytes. We show that increasing periods of 56 °C exposure of sperm produces a progressive loss of Ca(2+) oscillations after ICSI. The decrease in Ca(2+) oscillations produces a reduction in oocyte activation and embryo development to the blastocyst stage. We treated such oocytes that failed to activate after ICSI either with Ca(2+) ionophore, or with Sr(2+) media which causes Ca(2+) oscillations, or we injected them with recombinant human PLCζ. All these treatments rescued oocyte activation, although Sr(2+) and PLCζ gave the highest rates of development to blastocyst. When recombinant PLCζ was given to oocytes previously injected with control sperm, they developed normally to the blastocyst stage at rates similar to that after control ICSI. The data suggest that recombinant human PLCζ protein is an efficient means of rescuing oocyte activation after ICSI failure and that it can be effectively used even if the sperm already contains endogenous Ca(2+) releasing activity.

Topics: Animals; Blastocyst; Calcium Signaling; Disease Models, Animal; Embryonic Development; Female; Fertilization; Hot Temperature; Humans; Infertility, Male; Ionomycin; Ionophores; Male; Mice; Mice, Inbred Strains; Oocytes; Phosphoinositide Phospholipase C; Recombinant Fusion Proteins; Sperm Injections, Intracytoplasmic; Strontium

Does the application of three different artificial activating stimuli lead to a difference in pre- and post-implantation embryo development in the wobbler mouse, a mouse model with oocyte activation deficient round-headed sperm cells similar to human globozoospermia?. No gross differences were found between strontium chloride, electrical pulses or ionomycin with respect to the pre- and post-implantation development in the wobbler mouse.. Fertilization failure following intra-cytoplasmic sperm injection (ICSI) occurs in 1-3% of the ICSI cycles in human assisted reproduction technology (ART) and has been successfully overcome by different artificial activating stimuli. No comparison has been made yet in terms of their efficiency and safety.. Calcium release and embryo development were compared between oocytes fertilized by wobbler and wild-type (WT) sperm following ICSI with or without three different artificial activating agents. Preimplantation development was assessed on 70 injected oocytes on average per group. On average, 10 foster mothers were used per activating group to compare post-implantation development.. We used the wobbler mouse model that possesses oocyte activation deficient round-headed sperm cells. First, the calcium release following ICSI using wobbler sperm was compared with that of WT sperm. Outcome measures were the percentage of oocytes that showed calcium release and their mean amount of calcium rises. Secondly, the pre- and post-implantation development was assessed following ICSI with wobbler sperm plus artificial oocyte activation using either: (i) strontium chloride (Wob-Sr), (ii) electrical pulses (Wob-E) or (iii) ionomycin (Wob-I). Outcome measures were the activation, cleavage and blastocyst rates and the assessment of blastocyst quality by differential staining. Following mouse embryo transfer, pregnancy and birth rates as well as mean litter sizes were examined. Finally, pups were followed up until 8 weeks of age and then mated with fertile controls to assess their fertility.. The percentage of oocytes showing calcium rises as well as the number of calcium rises per oscillating oocyte were significantly lower in the wobbler group when compared with the WT group (9.3 versus 96% and 2.1 calcium rises versus 31 calcium rises) (P < 0.001). The fertilization rate was significantly lower in the wobbler group (11.4%) when compared with the WT group (92.1%) and the artificial activation groups (strontium chloride: 99%, electrical pulses: 99% and ionomycin: 81%, respectively) (P < 0.001). Post-implantation development did not differ significantly between the WT and artificial activation groups, with pregnancy rates in favor of strontium chloride and electrical pulses. The weight of the male pups did not differ between the study groups, whereas the weight of the female pups originating from Wob-Sr embryos was significantly lower at weeks 2, 3 and 4 when compared with female pups originating from WT embryos. However, the latter difference was not observed at later time points, nor in the other artificial activating groups. All offspring mated successfully with fertile controls.. Results in animal models should be extrapolated with caution to a subfertile human population. Also, ionomycin is currently the most widely used artificial oocyte activating agent in human ART.. The low frequency of observed calcium rises and the low activation rate make the wobbler mouse a highly suitable model to study oocyte activation deficiency. Strontium chloride and electrical pulses were more efficient means to restore fertilization rates and to support pre- and post-implantation embryonic development than ionomycin.. This work was supported by the Flemish foundation of Scientific Research (FWO-Vlaanderen) (aspirant clinical research mandate to F.V.M., fundamental clinical research mandate to P.D.S.); and Ghent University grant (KAN-BOF E/01321/01 to B.H.). The authors have no competing interests to declare.

Topics: Animals; Blastocyst; Calcium; Disease Models, Animal; Electrophysiology; Embryo Implantation; Embryo Transfer; Female; Infertility, Male; Ionomycin; Male; Mice; Oocytes; Pregnancy; Pregnancy, Animal; Sperm Injections, Intracytoplasmic; Spermatozoa; Strontium

Failed fertilization after intracytoplasmic sperm injection (ICSI) can be due to a reduced oocyte-activation capacity caused by reduced concentrations and abnormal localization of the oocyte-activation factor phospholipase C (PLC) zeta. Patients with this condition can be helped to conceive by artificial activation of oocytes after ICSI with calcium ionophore (assisted oocyte activation; AOA). However some concern still exists about this approach. Mouse models could help to identify potential oocyte-activation strategies and evaluate their safety. In this study, the fertilizing capacity of wobbler sperm cells was tested and the efficiency of AOA with two exposures to ionomycin to restore fertilization and embryo development was studied. The quality of the obtained blastocysts was assessed and embryo transfer was performed to evaluate post-implantation development. The presence of PLCzeta in the spermatozoa and testis of the wobbler mouse was evaluated by PLCzeta immunostaining and quantitative reverse-transcription polymerase chain reaction. Sperm cells from wobbler mice had reduced fertilizing capacity and abnormalities in PLCzeta localization, but not in its expression. Artificially activating the oocytes restored fertilization and embryo development. Therefore, the wobbler mouse can be a model for failed fertilization after ICSI to study PLCzeta dynamics and aid in optimization of the AOA method.

Topics: Animals; Embryo Transfer; Fertilization; Infertility, Male; Ionomycin; Male; Mice; Oocytes; Phosphoinositide Phospholipase C; Reverse Transcriptase Polymerase Chain Reaction; Sperm Injections, Intracytoplasmic; Spermatozoa; Treatment Failure

To report a successful pregnancy after intracytoplasmic sperm injection (ICSI) with artificial oocyte activation (AOA) on a patient whose fertilization rate after ICSI was extremely low; and to report on cytologic analyses of the fertilization failure (FF) eggs after ICSI and a biologic assessment of the sperm of this patient.. Case report with an assessment of gamete function.. University hospital and an experimental laboratory.. A couple with severe oligoasthenozoospermia, whose seventh attempt at ICSI ended in the failure.. Cytologic analyses of FF eggs after ICSI, AOA after ICSI, and analyses of human sperm oocyte activation ability and centrosomal function.. Fertilization arrest after ICSI was observed in FF eggs at various stages of fertilization. After artificial oocyte activation by exposure to ionomycin, clinical pregnancy was confirmed, and a healthy baby was born. As assessed by heterologous ICSI of human sperm into bovine oocytes, there was no significant difference in the oocyte activation rates between the patient's and control sperm, but the sperm centrosomal function was low in the patient's sperm (48.5% vs. 69.6%).. We report a successful pregnancy after ICSI with AOA using a calcium ionophore, after critical cytologic analyses of the FF eggs. Furthermore, sperm centrosomal function was low, indicating that sperm's ability to process the events of fertilization after the oocyte activation was poor in this patient.

Topics: Adult; Animals; Biological Assay; Calcium; Cattle; Centrosome; Embryo Culture Techniques; Embryo Transfer; Female; Humans; Infant, Newborn; Infertility, Male; Ionomycin; Ionophores; Live Birth; Male; Oocyte Retrieval; Oocytes; Ovulation Induction; Pregnancy; Sperm Injections, Intracytoplasmic; Sperm-Ovum Interactions; Spermatozoa; Treatment Failure

To determine if a low acrosin activity in otherwise normal ejaculate from infertile patients is associated with an impairment of sperm functions involved in the hamster egg penetration.. A tertiary care center, the Andrology Clinic, Department of Internal Medicine, University of L'Aquila.. Nine infertile patients with low acrosin activity in otherwise normal ejaculate (including normal immunoreactivity for acrosin) were studied; nine fertile men served as a control group. The two groups were homogeneous for seminal parameters.. The hamster egg penetration assay and the acrosome reaction rate assessment were performed on capacitated sperm suspensions in basal conditions and after ionophore challenge.. The penetration rate, the penetration index, and the acrosome reaction rate were measured and compared between the groups.. No penetration was achieved in seven patients and a low penetration was achieved in two cases. The difference with the control group was significant. The ionophore challenge was associated to penetration of hamster eggs in seven of nine patients, but the penetration index was significantly lower than the controls. Acrosin activity was correlated to hamster egg penetration. Both spontaneous and induced acrosome reaction rate were similar in the two groups.. A low sperm acrosin activity in otherwise normal ejaculate is associated with an impaired hamster egg penetration. This impairment does not seem to be due to altered dynamics of acrosome reaction.

Topics: Acrosin; Acrosome; Animals; Cricetinae; Female; Fluorescent Antibody Technique; Humans; Infertility, Male; Ionomycin; Male; Pregnancy; Sperm Capacitation; Sperm-Ovum Interactions; Spermatozoa