R. Santucci

Effect of sperm morphology and motile sperm count on outcome of intrauterine insemination in oligozoospermia and/or asthenozoospermia * †

Eighty-six couples with long-standing infertility and poor postcoital test, due to oligozoospermia and/or asthenozoospermia (68 cases) or mucus hostility (18 cases), were treated by 411 intrauterine inseminations (IUI) with motile sperm suspensions from the husbands semen. The pregnancy rate per couple in the group with abnormal semen was lower than in the group with mucus hostility (22% versus 38.9%). Influence of seminal and other parameters on outcome of IUI was assessed by discriminant analysis, and a significant correlation with pregnancy rate was found for motile sperm count and sperm morphology. Teratozoospermia (normal morphology less than 50%) affected the outcome of IUI both when associated with moderate oligozoospermia and/or asthenozoospermia (motile sperm count greater than or equal to 5 X 10(6)/mL) (success rate per couple: 11.1%), and, even more, when associated with severe oligozoospermia and/or asthenozoospermia (motile sperm count less than 5 X 10(6)/mL), where no pregnancy was achieved. In the absence of teratozoospermia, the success rate per couple both in severe and in moderate oligozoospermia and/or asthenozoospermia had similar results (33.3% versus 35.7%). In conclusion, the absence of teratozoospermia appears to be an effective criterion for selecting couples with infertility due to oligozoospermia and/or asthenozoospermia who may benefit from IUI.

Nitric Oxide Synthase Inhibition in Human Sperm Affects Sperm-Oocyte Fusion but Not Zona Pellucida Binding

Abstract There is recent evidence that mouse and human spermatozoa contain constitutive nitric oxide synthase (cNOS) and can synthesize nitric oxide. The aim of this study was to investigate whether the inhibition of human sperm cNOS could affect sperm-oocyte fusion and sperm binding to the zona pellucida (ZP). NG-nitro-l-arginine methyl ester (l-NAME) was used as cNOS inhibitor. Sperm-oocyte fusion was evaluated using the hamster egg penetration test (HEPT). The ZP binding was evaluated using the hemizona assay. l-NAME added from the onset of capacitation strongly inhibited sperm-oocyte fusion. This inhibitory effect was dose dependent, stereospecific, and suppressed by l-arginine in a dose-dependent manner. l-NAME also inhibited sperm-oocyte fusion in the HEPT enhanced with progesterone (P), where P (5 μM) was added for 15 min to capacitated sperm. A lesser but significant inhibition was also observed when sperm suspensions were exposed to l-NAME following capacitation in both versions of HEPT. On the contrary, l-NAME did not affect ZP binding. In conclusion, the present study provides the evidence that cNOS plays a role in the human sperms capacity to fuse with oocyte but not in the ZP binding.

Failure of intrauterine insemination in male immunological infertility in cases in which all spermatozoa are antibody-coated * †

OBJECTIVE To determine if the overcoming of the cervical mucus barrier removes the interference of sperm-bound antibodies with fertility. DESIGN Prospective case series. SETTINGS University-based intrauterine insemination (IUI) homologous program. PATIENTS Nineteen patients with all spermatozoa in the ejaculate coated by antisperm antibodies. As control group, 86 consecutive patients without antisperm antibodies, treated for oligoasthenozoospermia or mucus hostility. INTERVENTIONS Intrauterine inseminations (at least 3 attempts per couple). MAIN OUTCOME MEASURES The outcome of IUIs, demographic, and seminal parameters were compared between the two groups. RESULTS No pregnancy occurred in the couples with male immunological infertility, treated by 110 IUIs. Twenty-three pregnancies occurred in 22 (25.6%) of the control group couples who were treated by 411 IUIs. In the group of patients without antisperm antibodies, we demonstrated that the pregnancy rate (PR)/couple in oligoasthenozoospermia without teratozoospermia was similar to that achieved in normozoospermia (35% versus 38.9%), whereas it was significantly affected by teratozoospermia (3.6%). Only three patients with antisperm antibodies had teratozoospermia. Comparing the PR per couple and per cycle between the two groups of patients (with and without antisperm antibodies), excluding the patients with teratozoospermia, significant differences resulted (P less than 0.005 and P less than 0.005, respectively). The motile sperm count was not significantly different between the two groups, which also resulted to be homogeneous for demographic data. Moreover, the motile sperm count was not different between the patients with and without antisperm antibodies, who had successful IUI. CONCLUSIONS The analysis of this trial suggests that the failure of IUI in the treatment of male immunological infertility is imputable to antisperm antibodies when they involve all spermatozoa, regardless of semen quality.

Dynamics of the Global Tyrosine Phosphorylation During Capacitation and Acquisition of the Ability to Fuse with Oocytes in Human Spermatozoa

Abstract Phosphorylation of tyrosine residues in cellular proteins represents a major event during sperm capacitaton, but its relationship with the acquisition of sperm-fertilizing ability is still unclear. In this study we explored the relationship between the kinetics of the global tyrosine phosphorylation, monitored with a flow cytometric assay, and the acquisition of the human sperm ability to fuse with oocytes, evaluated with the progesterone-enhanced hamster egg penetration test. Sperm tyrosine phosphorylation appeared to be an early event in the capacitation process, with a 3.6-fold mean increase within 1 h of capacitation, but at this time sperm-oocyte fusion was extremely poor compared with that observed at 5 h of capacitation. Capacitation in calcium-free medium produced a 2-fold mean increase in tyrosine phosphorylation compared with that seen in complete capacitation medium both at 1 h and 5 h of capacitation, whereas sperm-oocyte fusion significantly increased only at 1 h, remaining unchanged at 5 h of capacitation. The cAMP analog, N,2-O-dibutyryladenosine 3′,5′-cyclic monophosphate (dbcAMP), prevented the inhibitory effect of seminal plasma on tyrosine phosphorylation but not on sperm-oocyte fusion. In conclusion, these results suggest that the acquisition of sperm-fertilizing ability is always associated with an increase of the global tyrosine phosphorylation, but tyrosine phosphorylation does not necessarily reflect the acquisition of the sperm-fertilizing ability. Flow cytometry assay, a reliable technique to quickly quantify the global levels of the human sperm tyrosine phosphorylation, could be useful for a further elucidation of the biological meaning of this process, with the perspective of its clinical use as a measure of the sperm-fertilizing potential.

Effect of Sperm‐Antibodies on Acrosome Reaction of Human Sperm Used for the Hamster Egg Penetration Assay

ABSTRACT: The effect of anti‐sperm antibodies (ASA) on the rate of acrosome reactions (AR) during “in vitro” capacitation of human sperm used for the hamster egg penetration assay (HEPA) was assessed. Motile sperm suspensions from donors were exposed to several sera and seminal plasma with sperm head‐directed ASA, then they were washed and capacitated “in vitro.” After capacitation, the proportion of acrosome‐reacted viable sperm was assessed by staining with Fluoresceinated Pisum Sativum Agglutinin and supravital stain Hoechst 33258. ASA of any immunoglobulin class did not significantly affect either the AR rate, or the hamster egg penetration rate. In conclusion, interference of ASA on spontaneous AR rate during “in vitro” capacitation can not be advocated as an explanation of the impairment of the interaction of human sperm with egg or its vestments, which have been reported in several studies.

Occurrence of the Interference of Sperm-Associated Antibodies on Sperm Fertilizing Ability as Evaluated by the Sperm-Zona Pellucida Binding Test and by the TEST-Yolk Buffer Enhanced Sperm Penetration Assay

PROBLEM: This study was performed to evaluate the occurrence as well as the level of the interference of sperm‐associated antibodies on fertilization process.

Effect of ionophore challenge on hamster egg penetration and acrosome reaction of antibody-coated human sperm.

PROBLEM: Following the demonstration that antisperm antibodies do not affect the spontaneous acrosome reactions (AR) of human sperm used for the hamster egg penetration assay (HEPA), we evaluated the effect of the ionophore challenge on HEPA and AR of antibody‐coated sperm.

Intrauterine insemination with or without mild ovarian stimulation in couples with male subfertility due to oligo/astheno- and/or teratozoospermia or antisperm antibodies: a prospective cross-over trial.

Seventy-three couples with male subfertility, which was due to oligo/astheno- and/or teratozoospermia (n = 63) or antisperm antibodies (n = 10), were randomly assigned to sequential timed natural intercourse, intrauterine insemination (IUI) and IUI + mild ovarian hyperstimulation. From the analysis of 384 observed cycles, IUI was shown to be effective in oligo/asthenozoospermia without severe teratozoospermia, when it was associated with moderate multifollicular induction, and in male immunologic subfertility, IUI was highly effective in nonstimulated cycles also.

Modification of the slide agglutination test for the detection of sperm-agglutinins.

Modifikation des Objektträger‐Agglutinationstests zur Bestimmung der Sperma‐Agglutinine

Detection of Sperm Surface Related Antibodies by Indirect Immunofluorescence Test on Sperm Suspensions. Indirect I FT on Sperm Suspensions

Summary:  An indirect immunofluorescence test (IIFT) using sperm suspensions was carried out on 20 sera with sperm agglutinins (SA) and on 25 negative controls. IIFT gave results highly correlated with the occurrence of sperm agglutinating activity. Moreover, a relation was found among class of Ig involved in IF reactivity, fluorescent stain pattern and type of sperm agglutinations. In all sera with “mixed” or “tail‐tail” sperm agglutinating activity, IgG were involved in IF reactivity; the fluorescent stain constantly appeared in a granular pattern along the sperm tail and most often on the head surface too. In relation to high titres of “head‐head” sperm agglutinating activity, IgM were involved in IF reactivity; here the fluorescent stain appeared to be localized on acrosomal surface. The results indicate that indirect IFT on sperm suspensions specifically detects sperm surface related antibodies.