calcimycin and Infertility--Male

Are pregnancy and birth rates affected by artificial oocyte activation (AOA) with SrCl2 or calcimycin after ICSI for couples with male-factor infertility linked to abnormal sperm morphology or for couples with previous ICSI cycles of unexplained low fertilization or inadequate fertilization associated with impaired oocyte morphology?. AOA with either SrCl2 or calcimycin can improve the rates of clinical pregnancy, ongoing pregnancy and live birth compared with ICSI alone, and the two agents have diverse effects for different subgroups of patients.. ICSI is a successful treatment for infertility, but not in all individuals. AOA has potential to overcome inadequate fertilization in ICSI. Calcimycin and SrCl2 are candidate agents for AOA, but their effectiveness remains to be compared.. This study was a randomized, open-label, three-arm, parallel-group, double-centre, superiority trial conducted between April 2015 and January 2016. The study evaluated the effects of AOA with calcimycin or SrCl2 for clinical pregnancy rates after ICSI and included 343 couples divided into three groups.. Couples were included if they had two previous ICSI cycles of no or low fertilization (0-30%) with unknown causes or impaired oocyte morphology. Male-factor infertility cycles (frozen-thawed sperm, surgically retrieved sperm or ejaculates contained <10 millions spermatozoa/ml) undergoing their first ICSI attempt were also included if they had 100% abnormal sperm morphology (including globozoospermia and tapered-head). Couples were randomized to undergo ICSI with SrCl2 AOA, ICSI with calcimycin AOA or ICSI alone, with clinical pregnancy as the primary endpoint. Effect sizes were summarized as absolute rate differences (ARDs) and odds ratios (ORs), with precision evaluated by 95% CIs.. Both SrCl2 and calcimycin AOA improved clinical pregnancy rates compared to ICSI alone (49, 42 and 27%; ARD 22, 95% CI: 9-33; P = 0.0007 and ARD 16, 95% CI: 3-27; P = 0.014). SrCl2 and calcimycin AOA were also superior to ICSI alone on the rates of ongoing pregnancy (42, 36 and 23%; P = 0.0019 and P = 0.023) and live birth (40, 33 and 18%; P = 0.0002 and P = 0.012). Among couples with previous ICSI cycles of low fertilization, AOA with SrCl2 (but not with calcimycin) was superior to ICSI alone for rates of clinical pregnancy (ARD 35 percentage points (pp), P = 0.0007), ongoing pregnancy (ARD 27 pp, P = 0.009) and live birth (ARD 37 pp, P = 0.002). Among couples affected by male-factor infertility, AOA with calcimycin (but not with SrCl2) was superior to ICSI alone for rates of clinical pregnancy (ARD 22 pp, P = 0.006), ongoing pregnancy (ARD 19 pp, P = 0.013) and live birth (ARD 17 pp, P = 0.02).. This study was an open-label trial, and this design might have introduced bias, although randomization methods were used. The study did not include a longitudinal follow-up, so further evidence is required to demonstrate the safety of AOA.. The decision to use SrCl2 or calcimycin for AOA after ICSI may depend on whether the activation failure originates in the oocyte or the sperm.. The study received no funding and the authors have no conflicts of interest to declare.. NCT02424214.. 22 April 2015.. 27 April 2015.

Topics: Adult; Birth Rate; Calcimycin; Female; Humans; Infertility, Male; Male; Oocyte Retrieval; Oocytes; Pregnancy; Pregnancy Rate; Sperm Injections, Intracytoplasmic; Strontium

To investigate the pathogenesis of globozoospermia, fertilization ability of round-headed sperm, and the application value of assisted oocyte activation in intracytoplasmic sperm injection (ICSI) for the wives of glohozoospermia men.. We collected oocytes from the wives of 2 globozoospermia patients and randomly divided them into two groups after ICSI to receive calcium ionophore A23187-activation and conventional treatment, respectively. We reviewed the relevant literature published at home and abroad, and discussed the etiology of globozoospermia, fertilization ability of round-headed sperm, and treatment options for this disease.. Quality embryos were obtained in the A23187-activation group while no fertilized oocytes, oocyte cleavage, quality embryos, or blastular formation were found in the conventional treatment group. Both women achieved pregnancy and gave birth to healthy neonates after transfer of the quality embryos from the A23187-activation group.. Calcium ionophore A23187 can be applied to ICSI for the wives of globozoospermia men and bring about desirable clinical outcomes. Meanwhile, attention should be paid to its safety.

Topics: Calcimycin; Calcium Ionophores; Female; Humans; Infertility, Male; Male; Oocytes; Pregnancy; Sperm Injections, Intracytoplasmic; Spermatozoa

To determine if fertilization antigen (FA)-1 will remove autoantibodies from the surface of sperm cells of immunoinfertile men by immune adsorption and permit an increased acrosome reaction (AR).. Prospective analytic study.. University medical center.. Men from 18 infertile couples with autoantibodies present on their spermatozoa.. Sperm samples after processing were examined for antibody binding and AR before and after adsorption with control medium or FA-1.. Sperm-bound antibody was assessed by the immunobead assay (immunoglobulin [Ig] A and IgG) and the AR by induction with ionophore A23187.. Adsorption with FA-1 compared with control medium increased immunobead-free swimming sperm an average of 50% and 76% for IgA and IgG antisperm antibodies, respectively, with 78% and 100% of the 18 semen specimens increasing significantly. The AR rate increased an average of 10.3% compared with control medium and showed improvement in 78% of the sperm samples after FA-1 adsorption.. The FA-1 sperm antigen appears to significantly free sperm cells coated with autoantibodies in the semen of most infertile men examined. Reducing sperm-bound antibodies that inhibited the AR allowed the sperm cells to undergo successful AR induction by calcium ionophore.

Topics: Acrosome Reaction; Antigen-Antibody Reactions; Antigens; Autoantibodies; Calcimycin; Humans; Immunoglobulin A; Immunoglobulin G; Immunosorbent Techniques; Infertility, Male; Ionophores; Male; Prospective Studies; Spermatozoa

To determine whether or not acrosome evaluation can enhance the prediction of IVF results when associated to conventional semen parameters.. Acrosome reaction, sperm concentration, motility, and morphology were recorded in 131 semen samples from patients undergoing an IVF attempt.. Spontaneous acrosome loss after a 24-hour incubation in B2 medium and after induction by calcium ionophore A23187 and phorbol ester, phorbol 12-myristate 13-acetate 4-O-methyl ether (TPA).. Statistically significant differences between fertilization failures and successes were found for concentration, viability, spontaneous and induced acrosome reaction, and most parameters of motility and morphology. However, none of the parameters could predict > 64% of IVF results when studied alone. A progressive discriminant analysis allowed to predict up to 83% of IVF results, by classifying sperms through their normal forms, rapid motility, spontaneous acrosome loss, enlarged heads, multiflagellar forms, vitality, linear motility, and acrosome response to TPA. The other parameters, including concentration and response to calcium ionophore, had no additive value.. The study of acrosome function, through spontaneous acrosome loss and response to TPA, is of great interest in clinical practice when associated to some parameters of motility and morphology. However, it appears that response to calcium ionophore, one of the most studied parameters, is of poor practical interest.

Topics: Acrosome; Calcimycin; Discriminant Analysis; Female; Fertilization in Vitro; Humans; Infertility, Female; Infertility, Male; Male; Semen; Sperm Capacitation; Sperm Count; Sperm Motility; Sperm-Ovum Interactions; Spermatozoa; Tetradecanoylphorbol Acetate; Treatment Outcome

To assess the utility of the acrosome reaction (AR) to ionophore challenge test in determining the sperm treatment protocols for patients undergoing assisted reproduction.. One hundred twenty-one couples undergoing an IVF-ET or GIFT procedure from January to July 1992 were included in this prospective study. All cases had a preliminary semen analysis within the previous 3 months and an AR to ionophore challenge test was carried out unless an acceptable fertilization rate occurred on previous IVF. For those patients whose AR to ionophore challenge score was below the accepted fertile range of > or = 10%, a second AR to ionophore challenge test was performed after exposure of sperm to the stimulant pentoxifylline. Couples then were managed by assisted reproduction with randomized allocation of oocytes for fertilization with a standard sperm preparation or with added sperm stimulants, either 3.6 mM pentoxifylline alone or combined with 3.0 mM 2-deoxyadenosine. The study was double-blind with neither the patients nor the embryologist knowing the AR to ionophore challenge result at the time of the IVF procedure.. Data from the preliminary semen analyses and AR to ionophore challenge scores were correlated with the fertilization rates achieved using control and treated sperm preparations. The rates of total fertilization failure and the numbers of clinical pregnancies occurring in each subgroup were also recorded.. All AR to ionophore challenge groups showed normal sperm counts except the groups with poor AR to ionophore challenge, which demonstrated reduced sperm counts. The group with normal AR to ionophore challenge scores or previous normal fertilization showed satisfactory fertilization rates with either control or treated sperm, although some individual cases showed reduced fertilization with treated sperm. The fertilization rate for the group with low AR to ionophore challenge scores improved significantly with pentoxifylline, and the benefit was greatest when this had been predicted from the AR to ionophore challenge studies. Cases with persisting poor AR to ionophore challenge despite pentoxifylline showed no significant improvement in fertilization rates with sperm exposed to either sperm stimulant regimens. Poor AR to ionophore challenge scores were also predictive of total fertilization failure, but this problem was reduced by sperm stimulation. The AR to ionophore challenge score at 10% cutoff level showed optimal levels of sensitivity (82.1%), highest negative predictive value (82.1%), and lowest false negative rate (17.9%).. The AR to ionophore challenge test is useful in the assessment and management of the male factor in assisted reproduction. It can be used to identify the majority of cases who will benefit from the use of sperm stimulants.

Topics: Acrosome; Calcimycin; Cell Membrane Permeability; Deoxyadenosines; Double-Blind Method; Female; Fertility; Fertilization; Fertilization in Vitro; Gamete Intrafallopian Transfer; Humans; Infertility, Male; Male; Pentoxifylline; Prospective Studies; Reproduction; Spermatozoa

Intracytoplasmic sperm injection (ICSI) is commonly used to treat severe male factor infertility in assisted reproduction. A small percentage of patients face suboptimal fertilisation rate or even fertilisation failure despite having ICSI. Artificial oocyte activation (AOA) has been proposed as a suitable method to overcome their problem. This is a retrospective cohort analysis of ICSI cycles undergoing AOA. Injected metaphase II oocytes were exposed to either calcium ionophore (A23187) after ICSI or injection of calcium chloride during ICSI followed by incubation with A23187 after ICSI. The previous ICSI cycles of the patients formed the historical control group. Thirty-four AOA cycles were analysed. The normal fertilisation rate (52.1%) was significantly improved in the AOA group. The percentage of failed fertilisation cycles (11.8%) were significantly reduced in the AOA group. The cumulative clinical pregnancy rate (47.1%) and live birth rate (29.4%) were significantly increased when compared to the previous cycles. Subgroup analysis revealed that the performance of the A23187 only protocol and the concomitant injection of calcium chloride protocol were comparable in terms of laboratory parameters and pregnancy outcomes. AOA is an effective method to improve the fertilisation rate and pregnancy outcome of infertile couples with previous fertilisation problem after ICSI.IMPACT STATEMENT

Topics: Calcimycin; Calcium Chloride; Female; Fertilization in Vitro; Humans; Infertility, Male; Male; Oocytes; Pregnancy; Pregnancy Rate; Retrospective Studies; Sperm Injections, Intracytoplasmic

Progesterone (P

Topics: Acrosome; Acrosome Reaction; Adult; Calcimycin; DNA Fragmentation; Feasibility Studies; Female; Fertilization in Vitro; Humans; Infertility, Male; Male; Predictive Value of Tests; Pregnancy; Pregnancy Rate; Progesterone; Prognosis; Sensitivity and Specificity; Sperm Motility; Spermatozoa; Treatment Outcome

Given the importance of the chaperone Heat Shock Protein A2 (HSPA2) in the regulation of male fertility, this study aimed to identify and characterize additional proteins that may rely on the activity of this chaperone in human spermatozoa.. In view of the findings in this study we propose that angiotensin converting enzyme (ACE) and protein disulfide isomerase A6 (PDIA6) are novel interacting proteins of HSPA2 and that this multimeric complex may participate in key elements of the fertilization cascade.. The molecular chaperone HSPA2 plays a pivotal role in the remodelling of the sperm surface during capacitation. Indeed, human spermatozoa that are deficient in HSPA2 protein expression lack the ability to recognize human oocytes, resulting in repeated IVF failure in a clinical setting. Moreover, our recent work has shown that defective HSPA2 function induced by oxidative stress leads to the aberrant surface expression of one of its interacting proteins, arylsulfatase A, and thus contributes to a loss of sperm-zona pellucida adhesion.. Human spermatozoa were collected from fertile donors, capacitated and prepared for Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) analysis. Protein complexes resolved via BN-PAGE were excised and their constituents were identified using mass spectrometry. The interactions between ACE, PDIA6 and HSPA2 were then confirmed using immunoprecipitation and proximity ligation assays and the localization of these proteins was assessed in isolated spermatozoa and commercially available human testis tissue sections. Finally, pharmacological inhibition of ACE was performed to assess the role of ACE in human sperm capacitation.. Herein we have identified ACE and PDIA6 as potential HSPA2-interacting proteins and shown that this assemblage resides in membrane raft microdomains located in the peri-acrosomal region of the sperm head. Additionally, the surface expression of PDIA6, but not ACE, was shown to be dynamically regulated during sperm capacitation and, like that of previously characterized HSPA2-interacting proteins, this surface expression proved vulnerable to oxidative stress. In terms of the functional significance of this protein complex, pharmacological inhibition of ACE significantly reduced the ability of human spermatozoa to undergo an agonist induced acrosome reaction (P < 0.01).. While these results provide a descriptive analysis of the PDIA6/ACE/HSPA2 complex, this study provides the impetus for further investigation into the role of PDIA6 and ACE in human sperm function.. As our research group, and others, have shown that HSPA2 is compromised in the spermatozoa of men with oocyte recognition defects, the characterization of these HSPA2-interacting proteins provides important insight into the complexity of the cellular pathways that may be affected in the spermatozoa of infertile individuals.. Large scale proteomics data can be accessed through the Proteomics Identifications Database (PRIDE).. This work was supported by the National Health and Medical Research Council. Grant # APP1046346. The authors have no competing interests to declare.

Topics: Acrosome Reaction; Calcimycin; Fertility; Gene Expression Regulation; HSP70 Heat-Shock Proteins; Humans; Hydrogen Peroxide; Infertility, Male; Male; Membrane Microdomains; Peptidyl-Dipeptidase A; Protein Binding; Protein Disulfide-Isomerases; Protein Multimerization; Signal Transduction; Sperm Capacitation; Spermatozoa; Testis

Mammalian sperm acquire fertilizing capacity in the female tract in a process called capacitation. At the molecular level, capacitation requires protein kinase A activation, changes in membrane potential and an increase in intracellular calcium. Inhibition of these pathways results in loss of fertilizing ability in vivo and in vitro. We demonstrated that transient incubation of mouse sperm with Ca(2+) ionophore accelerated capacitation and rescued fertilizing capacity in sperm with inactivated PKA function. We now show that a pulse of Ca(2+) ionophore induces fertilizing capacity in sperm from infertile CatSper1 (Ca(2+) channel), Adcy10 (soluble adenylyl cyclase) and Slo3 (K(+) channel) KO mice. In contrast, sperm from infertile mice lacking the Ca(2+) efflux pump PMACA4 were not rescued. These results indicate that a transient increase in intracellular Ca(2+) can overcome genetic infertility in mice and suggest this approach may prove adaptable to rescue sperm function in certain cases of human male infertility.

Topics: Adenylyl Cyclases; Animals; Calcimycin; Calcium Channels; Calcium Ionophores; Disease Models, Animal; Fertilization; Fertilization in Vitro; Infertility, Male; Large-Conductance Calcium-Activated Potassium Channels; Male; Mice, Inbred C57BL; Mice, Knockout; Models, Genetic; Spermatozoa

A strong positive correlation exists between teratozoospermia and reactive oxygen species production, which in turn has negative effects on their in vitro fertilisation outcome. Our aim of this study was to determine potential protective effects of α-tocopherol on teratozoospermia motility, viability, acrosome reaction and DNA integrity after 1-h in vitro incubation. Teratozoospermic semen samples were obtained from 15 volunteers aged between 20 and 30 years after 3-5 days of sexual abstinence. Samples were washed, centrifuged and incubated in 37 °C and 5% CO(2) until sperm swimmed-up. Spermatozoa were counted in the supernatant and divided into four groups, each contained 2 × 10(6) sperm/ml(-1). Groups one to four were incubated for 1 h with Ham's F-10 solution as control group, 10 μm A23187, 40 μmα-tocopherol and 10 μm A23187 + 40 μmα-tocopherol respectively. The results indicated that α-tocopherol has ability to enhance teratozoospermia viability and motility, while there were no ameliorative effects on acrosome reaction and DNA fragmentation. A23187 induced acrosome reaction in teratozoospermia and α-tocopherol significantly diminished this effect. In conclusion, although α-tocopherol could improve teratozoospermia motility and viability, its effects on DNA integrity and acrosome reaction ability as supplementation IVF culture media are not obvious.

Topics: Acrosome Reaction; Adult; alpha-Tocopherol; Calcimycin; DNA Fragmentation; Humans; In Vitro Techniques; Infertility, Male; Male; Semen; Sperm Motility

To evaluate the effect of artificial oocyte activation (AOA) on intracytoplasmic sperm injection (ICSI) cycles using surgically retrieved sperm.. Laboratory study.. Fertility/assisted fertilization center.. Couples undergoing surgical sperm retrieval for ICSI (n = 204).. Application of calcium ionophore A23187 for AOA.. Cycles were divided into experimental groups according to the origin of the sperm used for injection and the type of azoospermia: [1] testicular sperm aspiration in nonobstructive-azoospermic patients (TESA-NOA group, n = 58), [2] TESA in obstructive-azoospermic patients (TESA-OA group, n = 48), [3] and percutaneous epididymal sperm aspiration in obstructive-azoospermic patients (PESA-OA, n = 98). For each experimental group, cycles where AOA was applied (subgroup: activation) were compared with cycles in which AOA was not applied (subgroup: control). The fertilization, high-quality embryo, implantation, and pregnancy rates were compared among the subgroups.. For patients undergoing TESA, AOA did not improve ICSI outcomes for either type of azoospermia. However, for cases in which the injected sperm were retrieved from the epididymis, a statistically significantly increased rate of high-quality embryos was observed with AOA.. Artificial oocyte activation may improve ICSI outcomes in azoospermic patients when epididymal, but not testicular spermatozoa, are injected.

Topics: Azoospermia; Biopsy, Needle; Brazil; Calcimycin; Epididymis; Female; Fertilization in Vitro; Humans; Infertility, Male; Informed Consent; Male; Oocyte Retrieval; Oocytes; Pregnancy; Sperm Injections, Intracytoplasmic; Sperm Retrieval

To present the effectiveness of diagnostic heterologous intracytoplasmic sperm injection (ICSI), mouse oocyte activation test (MOAT), and ICSI combined with assisted oocyte activation (AOA) in a globozoospermic patient.. A case report.. A private IVF center, Japan.. A patient with globozoospermia.. MOAT in a mouse and ICSI combined with AOA in a human.. Ultrastructure, MOAT, fertilization, and pregnancy.. The transmission electron micrographs showed 100% round-headed spermatozoa lacking an acrosome. MOAT showed that the fertilization rate was 68.4% (13/19) when AOA was used but 0% (0/19) when AOA was not used. After the diagnosis of globozoospermia and sperm-related activation deficiency, 17 human mature oocytes were activated with calcium ionophore after ICSI was performed. The fertilization rate was 88.2% (15/17), and 11 blastocysts were cryopreserved using the vitrification method to prevent severe ovarian hyperstimulation syndrome. A single vitrified-warmed blastocyst was transferred. A gestational sac with fetal heart movements was recognized, and a healthy boy weighing 3180 g was born at 40 weeks of gestation by cesarean section without any congenital abnormality.. MOAT allows discrimination between sperm- and oocyte-related fertilization failures and shows the effectiveness of AOA.

Topics: Adult; Animals; Biological Assay; Calcimycin; Calcium; Cesarean Section; Cryopreservation; Embryo Transfer; Female; Humans; Infertility, Male; Ionophores; Live Birth; Male; Mice; Microscopy, Electron, Transmission; Oocyte Retrieval; Oocytes; Ovulation Induction; Pregnancy; Sperm Injections, Intracytoplasmic; Sperm-Ovum Interactions; Spermatozoa

This study attempted to explain the mechanisms regulating boar fertility by examining seasonal changes in semen characteristics, the composition of seminal plasma and responsiveness of sperm acrosomes to Ca(2+) and the Ca(2+) ionophore A23187 (Ca(2+)/A23187). Sperm-rich and sperm-poor fractions were separately collected from 3 mature fertile Large White boars once a month over a one-year period. During the period of study, ambient temperature and relative humidity were recorded for within the stall in which the boars were kept and the semen characteristics, composition of the seminal plasma of sperm-rich fractions, and occurrence of the acrosome reaction in response to Ca(2+) (3 mM)/A23187 (0.3 microM) were examined. The highest mean maximum and minimum ambient temperatures were recorded in August-September, whereas the lowest mean maximum and minimum ambient temperatures were recorded in December and January, respectively. There was a moderate peak in relative humidity from July to October. The lowest percentages of motile spermatozoa and of spermatozoa with intact acrosomes and highest percentage of spermatozoa with abnormal morphology and strongest agglutination were seen in August-September. The total protein and albumin concentrations were lowest in August-September. Testosterone levels increased gradually as day length decreased after the summer solstice (June) and peaked in October-November. The percentage of acrosome reactions in response to Ca(2+)/A23187 was highest with the quickest response in August-September, as shown by the shortest time required for 50% of relative acrosome reactions. The farrowing rates were lowest in these same 2 months. These results suggest that seasonal infertility in Large White boars may be due, at least in part, to a combination of low motility, abnormal morphology including acrosomal abnormality, and early occurrence of the acrosome reaction in response to stimulus, possibly resulting from a decrease in acrosomal stabilizing proteins in the seminal plasma during summer. These changes may be modulated by heat/humidity stress and/or photoperiod-regulated testosterone.

Topics: Acrosome Reaction; Albumins; Animals; Calcimycin; Calcium; Chlorides; Fertility; Infertility, Male; Ionophores; Male; Seasons; Semen; Sodium; Sus scrofa; Temperature

New methods are needed for rapid and sensitive assessment of sperm function. As the ability to fertilize an oocyte is acquired during the capacitation process, assessments of sperm function have to be performed under fertilizing conditions. In this study, we monitored the dynamics of the temporal response of sperm from ejaculates of both fertile and subfertile boars to capacitating conditions in vitro (responsiveness) by following the changes in the response to calcium ionophore treatment and in [Ca(2+)](i). The differences between individual males were also investigated. Ionophore-induced changes and increased intracellular calcium ion content in boar spermatozoa were found to progress as a function of time during incubation under capacitating conditions. After primary kinetic analysis, 120 min was chosen as the point in time for assessment of responsiveness. Intra-boar variability in responsiveness parameters was relatively high (variation coefficient CV>30%), especially in the response to ionophore treatment, indicating that an isolated test may be inadequate for the evaluation of sperm function. Despite this high variability, there were markedly significant individual differences with respect to changes during capacitation, and there were significant correlations between conventional and responsiveness sperm parameters. The population of samples from subfertile boars, was found to be heterogeneous in regard to sperm responsiveness to capacitating conditions. There were two significantly different classes of subfertile boars ("low" and "high" responders), indicating that fertility may be associated with suboptimal rather than maximal response (both too rapid and too slow membrane changes). Therefore, criteria for quality judgement should include both the low and upper limits of responsiveness. The use of responsiveness parameters together with conventional spermatological parameters improved the prediction level of multiple regression models for farrowing rate and litter size. It can be concluded that the combination of sperm responsiveness parameters applied here is a suitable tool for the evaluation of sperm function.

Topics: Acrosome Reaction; Animals; Calcimycin; Calcium; Cell Death; Infertility, Male; Insemination, Artificial; Ionophores; Kinetics; Male; Regression Analysis; Sperm Capacitation; Sperm Motility; Spermatozoa; Swine; Swine Diseases

Topics: Acrosome; Animals; Calcimycin; Horse Diseases; Horses; Infertility, Male; Ionophores; Male

Sperm preparation and acrosomic reaction have been largely studied to explain the physiology of sperm as reproductive cell.. To identify the morphological changes on in vitro prepared human sperm cells and in spontaneous and ionophore A23187 induced acrosome reaction of infertile males, through the application of the chlortetracycline test.. Experimental prospective study.. Sperm samples of 17 subfertile males were processed as routine for a sperm analysis, we carried out sperm preparation and induction of the acrosomic reaction by means of calcium ions A23187. The prepared sperm with direct swim-up was assessed at one hour and at four hours. Direct aliquots were taken and processed for the test with CTC, we considered as positive result obtaining more than 15% increase in the induced AR versus spontaneous.. The whole population was classified as asthenozoospermic. Eight training morphological patterns were identified and we found four types of responses for the induced AR: Type I had an increase at one and at four hours incubation (47%); type II had an increase only after 1 hour incubation (23.5%); type III had an increase only at 4 hours incubation (17.7%), and type IV did not have increase (11.8%).. These results suggest that in astenozoospermic patients the training process has variations during the incubation time, demonstrated by the response at A23187. These findings could let us to select better spermatozoa in this group of males at the moment when capacitance is actually attained, thus being able to carry out more effective techniques in assisted reproduction.

Topics: Acrosome Reaction; Calcimycin; Chlortetracycline; Humans; Infertility, Male; Ionophores; Male; Spermatozoa

To describe a successful pregnancy and delivery from frozen-thawed embryos after intracytoplasmic sperm injection (ICSI) and assisted oocyte activation in a globozoospermic patient with mosaic Down syndrome.. Controlled clinical study.. IVF Laboratory, PL Infertility Clinic, Seoul, Korea.. A couple with infertility resulting from globozoospermia with mosaic Down syndrome: 47,XY,+21[7]/46,XY[33].. Semen analysis, karyotyping, ICSI, assisted oocyte activation, assisted hatching, and frozen-thawed ET.. Fertilization rate, implantation, pregnancy, and delivery.. Thirty-eight oocytes were aspirated, and round-headed spermatozoa were injected into 35 oocytes in metaphase II. Assisted oocyte activation with calcium ionophore A23187 after ICSI resulted in a high fertilization rate (21 of 35, 60%; 2 pronuclei in 18 of 21; 3 pronuclei in 3 of 21) and good embryo development. At 3 days after ICSI, 5 embryos of good quality were surgically transferred to the endometrium after assisted hatching, but no pregnancy occurred. After 2 months, the surgical transfer of 4 frozen-thawed embryos after assisted hatching led to an ongoing pregnancy. A female infant weighing 3,000 g was delivered at 38 weeks of gestation by cesarean section.. We report the first successful pregnancy and delivery from frozen-thawed embryos after ICSI and assisted oocyte activation in a globozoospermic patient with mosaic Down syndrome.

Topics: Adult; Calcimycin; Cesarean Section; Cryopreservation; Down Syndrome; Embryo Implantation; Embryo Transfer; Embryo, Mammalian; Female; Humans; Infertility, Male; Ionophores; Male; Mosaicism; Oocytes; Pregnancy; Pregnancy Outcome; Sperm Head; Sperm Injections, Intracytoplasmic; Spermatozoa

The aim of this study was to determine the relationship between calcium ionophore A23187-induced acrosome reaction (AR) and sperm fertilizing ability. Semen samples remaining after preparation for standard IVF were studied in 109 patients who had sperm concentrations > or =20 x 10(6)/ml. Ionophore-induced AR was performed on motile spermatozoa selected by centrifugation on a Percoll gradient. Semen analysis was performed using standard methods. Patients with higher (>50%, n = 76) fertilization rates had significantly higher ionophore-induced AR than patients with lower (<50%, n = 33) fertilization rates (49 +/- 14 versus 38 +/- 21%, P < 0.05). When the data from all patients were analysed by logistic regression, only the percentage sperm motility in insemination medium and ionophore-induced AR were significantly related to fertilization rates. Similar results were also obtained when the data from a subgroup of patients with poor (<15% normal) sperm morphology were analysed. However, when patients with normal sperm morphology > or =15% were analysed separately, only sperm count and the percentage of spermatozoa with progressive motility in semen were significantly related to fertilization rates. In conclusion, ionophore-induced AR was significantly related to fertilization rates in vitro mainly in patients with teratozoospermic semen. Tests for ionophore-induced AR may provide additional information about sperm fertilizing ability but may not indicate specific defects of the physiological AR.

Topics: Calcimycin; Female; Fertilization; Fertilization in Vitro; Humans; Infertility, Male; Ionophores; Male; Semen; Sperm-Ovum Interactions; Spermatozoa; Treatment Failure

To examine the outcome of intracytoplasmic sperm injection (ICSI) with round-headed sperm (globozoospermia).. Retrospective analysis.. In vitro fertilization laboratory with extensive ICSI experience.. A patient couple with infertility because of globozoospermia seeking ICSI treatment.. Fertilization, cleavage, and pregnancy rates.. Intracytoplasmic sperm injection and calcium ionophore.. This couple experienced only 7% fertilization after ICSI in their first cycle. Treatment of the unfertilized oocytes with calcium ionophore 20 hours after ICSI-induced fertilization and cleavage of 70% of the oocytes. Embryo quality was fair to good. On the second cycle, 8 of the injected oocytes were treated with ionophore immediately after ICSI and the remaining 20 oocytes were untreated. Normal fertilization was achieved in 75% of the treated and 10% of the untreated oocytes. Treatment of these unfertilized oocytes with ionophore 20 hours after ICSI resulted in fertilization in 73%. Pregnancy was not achieved after either ICSI cycle. Ultrastructural analysis indicated multiple structural abnormalities in the sperm.. These results indicate that the round-headed sperm from this patient were incapable of oocyte activation after ICSI. This may be the reason for the frequent ICSI fertilization failure seen with this condition. Current ICSI procedures may not always overcome the infertility associated with globozoospermia, and further study of the etiology of this condition is needed.

Topics: Adult; Calcimycin; Female; Fertilization in Vitro; Humans; Infertility, Male; Ionophores; Male; Microinjections; Oocytes; Retrospective Studies; Spermatozoa

To investigate a method of assisted activation of human oocytes for the treatment of infertility resulting from globozoospermia associated with deficient oocyte activation capacity.. The mouse oocyte activation test was used to analyze the oocyte activation capacity of the sperm cells of a patient with globozoospermia. Fresh donor human oocytes were used for determining the most appropriate procedure for oocyte activation.. Infertility Center, University Hospital of Ghent.. A couple with infertility resulting from globozoospermia.. Intracytoplasmic sperm injection, assisted oocyte activation, and embryo transfer.. Oocyte activation and fertilization rates, implantation, and pregnancy.. Deficiency in oocyte activation capacity was found in the sperm of a patient with globozoospermia. This deficiency was proven by the mouse oocyte activation test and was confirmed further by lack of activation of human oocytes after simple sperm injection. Only human oocytes that were injected with sperm and subjected to calcium chloride and ionophore treatment underwent activation. Transfer of embryos obtained by this procedure of assisted oocyte activation resulted in an ongoing pregnancy.. Assisted oocyte activation of human oocytes is useful when globozoospermia is associated with absence of oocyte activation capacity in the sperm. These cases can be identified by the mouse oocyte activation test.

Topics: Adult; Animals; Calcimycin; Cytoplasm; Female; Fertilization in Vitro; Humans; Infertility, Male; Ionophores; Male; Mice; Microinjections; Oocyte Donation; Oocytes; Pregnancy; Spermatozoa

To study the effect of sperm-immobilizing antibodies from male sera on spontaneous and A23187-induced acrosome reactions (AR).. Swim-up spermatozoa obtained from three fertile donors were incubated with 13 sera with sperm-immobilizing antibodies obtained from infertile men and three control sera obtained from healthy fertile males. Sperm acrosomes were examined by staining with pisum sativum agglutinin labeled with fluorescein isothiocyanate (30 micrograms/ml; Sigma Chemical Co., St. Louis. MO) as spontaneous and A23187 (used at a final concentration of 10 microM; Sigma Chemical Co.) induced.. The incidence of spontaneous AR of spermatozoa incubated with antisperm antibody positive male sera (6.2 +/- 0.7) was significantly (P < 0.001) lower than that of spermatozoa incubated with control sera (10.7 +/- 0.5). And the incidence of A23187-induced and -inducible (incidence of induced minus spontaneous) ARs of spermatozoa incubated with sperm antibody-positive male sera (12.4 +/- 1.9 and 6.2 +/- 1.9) was significantly lower (P < 0.001) than that of spermatozoa incubated with control sera (31.0 +/- 0.5 and 20.3 +/- 0.9). Sperm-immobilizing antibody-positive sera decreased spontaneous, A23187-induced, and inducible ARs.. Sperm-immobilizing antibodies from male sera interfere with fertilization by inhibiting the AR.

Topics: Acrosome; Antibodies; Calcimycin; Humans; Infertility, Male; Male; Spermatozoa

It is believed that during the process of human fertilization, acrosome-intact spermatozoa bind to the surface of the zona pellucida which triggers the acrosome reaction and the enzymes released facilitate sperm penetration through the zona pellucida. We describe here reduced frequency of the acrosome reaction on the zona pellucida as a cause of infertility in 10 couples with long durations of infertility (average 6 years) and low (< 15%, n = 3) or zero (n = 7) fertilization rates in vitro. Sperm concentration, motility, velocity (Hamilton-Thorn), morphology and DNA normality were within the normal range in all the patients. Electron microscopy of spermatozoa did not reveal any specific ultrastructural defects. All couples were negative for antisperm antibodies by immunobead tests. Oocytes from other patients which failed to fertilize in in-vitro fertilization and normal donor spermatozoa were used as controls for sperm-zona pellucida binding and penetration experiments. Acrosome status of spermatozoa bound to the zona pellucida was assessed with a fluorescent lectin and electron microscopy. The mean number of spermatozoa bound to the zona pellucida was not significantly different between patients and controls. However, the acrosome reaction of spermatozoa bound to the zona pellucida after 2 h incubation was significantly lower (P < 0.001) in the patients (mean 5%, range 0-16) than in the controls (mean 68%, range 44-96). No zona pellucida (out of 40) was penetrated by patient spermatozoa whereas most (39/40) zonae were penetrated by control spermatozoa (average 27 spermatozoa/four zonae pellucidae).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Acrosin; Acrosome; Adult; Calcimycin; Female; Fertilization in Vitro; Humans; In Vitro Techniques; Infertility, Male; Male; Microscopy, Electron; Middle Aged; Sperm-Ovum Interactions; Zona Pellucida

To reveal the usefulness of Ca ionophore A23187-induced acrosome reactions (AR) in evaluating sperm fertilizing ability, the relationship between the percentage of AR and the results of the hamster test was examined in 65 men. Acrosome reacted spermatozoa were classified as type A (with FITC-PSA stain at the equatorial segment) or type B (without stain) and as alive or dead by staining with Hoechst 33258. The main results were as follows. 1. During the insemination period of 3 hours with zona-free hamster oocytes, sperm motility and the percentage of live type A spermatozoa (%AR) decreased significantly (p < 0.01). 2. Among the parameters examined, the %AR in the inseminating suspension exhibited the closest correlation with the sperm penetration rate (SPR) in the hamster test. 3. Among the 27 men with a %AR of less than 5%, 20 men (74.1) had SPRs less than 30%, indicating abnormal sperm fertilizing ability. In contrast, among the 19 men with %AR of 10% or more, 16 men (84.2%) had a SPR of 100%. These results suggest that the ionophore-induced acrosome reaction (AR) well predicts sperm fertilizing ability. We propose new criteria using %AR as follows: abnormal, < 5%; borderline, 5-10%; normal, > or = 10%.

Topics: Acrosome; Animals; Calcimycin; Cricetinae; Female; Fertilization; Humans; Infertility, Male; Male; Sperm Motility; Sperm-Ovum Interactions; Spermatozoa

To study the molecular origin and functionality of the plasma membrane of round-head spermatozoa in the human.. Clinical and laboratory study.. Patients in a clinical and academic environment.. Men with round-head spermatozoa.. Pisum sativum lectin homogeneously stains the surface of round sperm; however, the staining pattern and transmission electron microscopy show that the plasma membrane does not alter after exposure to the calcium ionophore A23187. In a clinical program, round-head spermatozoa injected subzonally into metaphase II oocytes with or without pretreatment with the fusogen polyethylene glycol did not bind or fuse to the oocyte surface.. The data suggests that plasma membrane fusion in human gametes is regulated by specific surface molecules and that exposure of these molecules on the sperm surface cannot be triggered by elevating intracellular calcium alone.

Topics: Adult; Calcimycin; Cell Membrane; Humans; Infertility, Male; Lectins; Male; Membrane Fusion; Microscopy, Electron; Microscopy, Electron, Scanning; Plant Lectins; Polyethylene Glycols; Sperm Motility; Spermatozoa

To evaluate the acrosome reaction and its prerequisite, a calcium influx, in spermatozoa of infertile men with a high incidence of abnormal sperm forms.. Prospective, controlled study.. Academic tertiary assisted reproduction center.. Patients (n = 14) were allocated in the study after semen evaluation showed teratozoospermia (< 14% normal sperm forms) as diagnosed by strict criteria.. After swim-up separation of the motile fraction, acrosome reactions were evaluated using Pisum sativum agglutinin (both spontaneously and exogenously induced with P and the calcium ionophore A23187, both at 10 microM); the intracellular-free [Ca2+]i was assessed by the fluorescent fura-2 indicator (basal and after P).. Patients did not show the typical P-induced wave of [Ca2+]i that was observed in controls but rather a blunted response, no response at all, or abnormal basal [Ca2+]i levels. The percent of basal acrosome reaction was significantly lower for patients versus controls postswim-up, and at 1 hour and 3 hours. Furthermore, there was a significant difference in the response of acrosome reaction to P both at 1 hour and 3 hours, with patients showing almost no response at all. However, patients' acrosome reaction response to the calcium ionophore was similar to those of fertile men.. Infertile patients with a high incidence of abnormal sperm forms as diagnosed by strict criteria have a low incidence of spontaneous acrosome reaction and a diminished P-stimulated acrosome reaction, whereas the nonspecific response to a calcium ionophore is conserved. Parallel abnormalities of [Ca2+]i were observed in patients, suggesting that these sperm populations may have a defective nongenomic P sperm receptor and/or abnormalities of other membrane transduction systems.

Topics: Acrosome; Calcimycin; Calcium; Humans; Infertility, Male; Male; Progesterone; Prospective Studies; Spermatozoa

The fertilizing potential of sperm depends on numerous properties which influence its functional competence. In order to understand the causes of IVF failure, we studied some of these properties-movement characteristics, hyperactivated motility, acrosome reaction-for infertile sperm (0% in vitro fertilization) and compared to those of fertile sperm (> or = 50% fertilization, control group). Movement characteristics, assessed by video-micrographic analysis, are significantly different for the 2 groups: infertile sperm register the lowest values for motility characteristics and hyperactivation. The addition of ionophore A23187 to the medium improves the hyperactivation and the acrosome reaction, the values for the latter, however, remain significantly lower compared to the control. These results are in favor of a detailed study of motility and induced acrosome reaction as complement for the diagnosis of male infertility.

Topics: Acrosome; Calcimycin; Female; Fertilization in Vitro; Humans; Infertility, Male; Male; Retrospective Studies; Semen Preservation; Sperm Count; Sperm Motility; Sperm-Ovum Interactions; Treatment Failure; Videotape Recording

To test whether pentoxifylline, a drug previously shown to sensitize spermatozoa from normal samples to the action of acrosome reaction stimuli, can be used to improve the fertilizing ability of spermatozoa from patients with acrosome reaction insufficiency.. Prospective analysis of pentoxifylline effects on the acrosome reaction and zona-free egg penetration; retrospective comparison of IVF results with and without the use of pentoxifylline.. Private hospital, public research center, and university-based laboratory.. In vitro fertilization patients selected on the basis of a previous acrosome reaction test.. None in the experimental part; IVF-ET in the clinical part of this study.. Frequency of the acrosome reaction, rate of penetration of zona-free eggs, normal fertilization in IVF attempts.. Pentoxifylline improves the acrosome reaction scores and zona-free egg penetration rates in patients with acrosome reaction insufficiency. Preliminary clinical experience shows an improvement of IVF results in these patients.. In vitro pentoxifylline treatment of spermatozoa to be used in IVF improves the sperm fertilizing ability in patients with acrosome reaction insufficiency. However, the effect of pentoxifylline on the acrosome reaction should be tested individually in each patient before the application of the drug in this new indication.

Topics: Acrosome; Calcimycin; Female; Fertilization in Vitro; Humans; Infertility, Male; Male; Pentoxifylline; Prospective Studies; Retrospective Studies; Sperm-Ovum Interactions; Spermatozoa

Spontaneous and ionophore-induced ability of spermatozoa to acrosome-react was examined in asthenozoospermic infertile patients and fertile donors. Spermatozoa were washed free of seminal plasma and capacitated in B2 medium for 2 h at 37 degrees C. Subsequently 5, 10, 20 and 30 microM A23187 (final concentrations) were added to equal aliquots of these samples and incubated for an additional 30 min. The acrosome reaction was then determined by the triple stain technique. The percentage of spontaneous reaction (no ionophore) in asthenozoospermic samples was similar to that in fertile samples (4.2 and 3.8, respectively). However, the ionophore-induced reaction rate remained significantly lower in asthenozoospermic samples than in normozoospermic samples.

Topics: Acrosome; Calcimycin; Humans; In Vitro Techniques; Infertility, Male; Male; Reference Values; Semen; Spermatozoa

Sperm obtained from groups of men with various semen profiles were incubated for 8 h in BWW medium containing human serum albumin to promote capacitation. Capacitation and the acrosome reaction were monitored by a chlortetracycline (CTC) fluorescence assay. Four distinct CTC patterns were observed on the sperm head. No significant difference was observed in the time-course curve of these CTC patterns in sperm obtained from normozoospermic, asthenozoospermic and oligozoospermic men. Spontaneous and A23187-induced acrosome reactions were also comparable in these groups. However, in sperm obtained from teratozoospermic and polyzoospermic men, the increase in CTC pattern associated with capacitation appeared slower and sluggish. In these two groups, the induced acrosome reaction was also significantly lower when compared to that in the other three groups of men. In polyzoospermia, the spontaneous acrosome reaction was significantly lower when compared to all the other groups. Fresh sperm would not undergo the acrosome reaction following A23187 treatment. The results of this study indicate sluggish (defective) capacitation and inability of capacitated sperm to undergo induced acrosome reaction in teratozoospermic and polyzoospermic men as evaluated by the CTC method.

Topics: Acrosome; Calcimycin; Chlortetracycline; Evaluation Studies as Topic; Humans; In Vitro Techniques; Infertility, Male; Male; Microscopy, Fluorescence; Semen; Sperm Capacitation

We have undertaken a prospective analysis of the diagnostic significance of three different criteria of human sperm function including the conventional semen profile, measurements of hamster-oocyte fusion, and determinations of reactive oxygen species generation in 139 couples. The latter, who were characterized by a lack of detectable abnormalities in the female partner, were followed up for a maximum of 4 years to determine the incidence of spontaneous pregnancy in the absence of therapeutic intervention. Assessments of monthly fecundity with life table analysis techniques revealed a highly significant, positive relationship between fertility and hamster-oocyte fusion rates that were measured in the presence of the ionophore, A23187. Conversely, reactive oxygen species generation was shown to be negatively associated with both the outcome of the sperm-oocyte fusion assay and fertility in vivo. The clinical significance of these diagnostic techniques was emphasized by the fact that within the same data set, the conventional semen profile was of no significant diagnostic value.

Topics: Animals; Calcimycin; Chronic Disease; Cricetinae; Female; Follow-Up Studies; Humans; Infertility, Male; Luminescent Measurements; Male; Oxygen Consumption; Pregnancy; Prognosis; Prospective Studies; Semen; Sperm-Ovum Interactions

Hyperactivated motility of capacitated sperm was studied before and after contact for 30 min with the Ca2+ ionophore A23187. Sperm from fertile donors and asthenozoospermic infertile patients were incubated in the presence of increasing concentrations of A23187 in the medium. At 5-10 microM, the ionophore induced a significant increase (P less than 0.05) in the percentage of hyperactivation of sperm from asthenozoospermic subjects. Higher concentrations (20 and 30 microM) were needed to enhance hyperactivation of sperm in the fertile population. In the latter, extended contact with the ionophore induced no significant change compared to a 30-min incubation period. Since this type of movement is an essential feature of the fertilizing gamete, a low hyperactivation rate may partly explain fertilization failure in the asthenozoospermic group.

Topics: Calcimycin; Fertility; Humans; Image Processing, Computer-Assisted; Infertility, Male; Male; Photomicrography; Sperm Motility; Statistics as Topic; Time Factors; Videotape Recording

The capacity to generate reactive oxygen species (ROS), both basally and after stimulation with the calcium ionophore A23187, was examined in the motile fraction of sperm isolated after swim-up from the semen of 10 naturally fertile men and three groups of infertile patients. The latter included: (1) men with a non-bacterial inflammation of the genital tract (n = 10); (2) men unable to impregnate their partners during an intra-uterine insemination programme (IUI) (n = 8) and their matched controls (n = 6); and (3) men with hypogonadotrophic hypogonadism (HH) who remained infertile after induction of spermatogenesis with gonadotrophin or gonadotrophin-releasing hormone therapy (n = 3) and their matched controls (n = 3). The levels of ROS production were elevated in the sperm of some infertile men with inflammation of the genital tract compared to those found in 10 naturally fertile men. In addition, sperm from those patients who remained infertile after an IUI programme produced higher amounts of ROS compared to their control group who became fertile. Similarly, the production of ROS by sperm from three patients with HH who remained infertile was significantly higher than those of the three men who became fertile. These data suggest that an excessive production of ROS by sperm may explain some cases of idiopathic male infertility.

Topics: Adult; Calcimycin; Cohort Studies; Humans; Hypogonadism; Infertility, Male; Luminol; Male; Oxygen; Prostatitis; Semen; Spermatozoa

In 1981, P. Talbot developed a triple-stain technique to estimate the number of human sperm undergoing normal acrosome reaction in fixed smears. In this method, live and dead sperm are first differentiated using the vital stain trypan blue. Sperm are then fixed in glutaraldehyde, dried onto slides, and the postacrosomal region and acrosome are differentiated using Bismark brown and Rose Bengal. Slides are examined at 1,000x with a bright-field microscope and assessed for the percentage of sperm that have undergone the normal acrosome reaction. Using this method we examined the time course of the acrosome reaction of human sperm incubated in mBWW the influence of human serum albumin, the calcium concentration of incubation media, Ca ionophore A 23187, and trypsin on the acrosome reaction of human sperm and the difference between the percentage of sperm undergoing normal acrosome reaction of fertile and infertile males. We got the following results: The percentage of human sperm undergoing acrosome reaction increased for the first six hours of incubation. Without human serum albumin the acrosome reaction did not occur. Ca ion could be one of the triggers of the acrosome reaction. Ca ionophore A23187 and trypsin induced the acrosome reaction in vitro. Sperm from oligozoospermic males, especially poorly motile sperm, could not undergo acrosome reaction so easily as sperm from fertile males could.

Topics: Acrosin; Acrosome; Calcimycin; Humans; In Vitro Techniques; Infertility, Male; Male; Oligospermia; Serum Albumin; Sperm Count; Sperm Motility; Spermatozoa; Staining and Labeling; Trypsin

Topics: Acrosome; Animals; Calcimycin; Cricetinae; Female; Humans; Infertility, Male; Male; Oocytes; Sperm Capacitation; Sperm-Ovum Interactions; Spermatozoa