RESUMO

The use of emergency contraception (EC) methods is increasing worldwide as it constitutes an effective way to prevent unplanned pregnancy after unprotected sexual intercourse. During the last decade, ulipristal acetate (UPA), a selective progesterone receptor modulator, has emerged as the most effective EC pill, and it is now recommended as first-line hormonal treatment for EC in several countries. Its principal mechanism of action involves inhibition or delay of follicular rupture, but only when administered during the follicular phase before the luteinizing hormone (LH) peak. However, considering the high efficacy of UPA, it is possible that it also exerts contraceptive effects besides ovulation. In the present review, we summarize and discuss the existing evidence obtained on the effect of UPA on sperm function and post-ovulatory events as potential additional mechanisms to prevent pregnancy. The bulk of evidence collected so far indicates that UPA would not affect gamete function; however, it could impair embryo-uterine interaction. Thus, besides the described effects on ovarian function, UPA contraceptive effectiveness might also be attributed to post-ovulatory effects, depending on the moment of the female cycle in which the drug is administered.

Assuntos

Anticoncepção Pós-Coito , Contraceptivos Hormonais/farmacologia , Norpregnadienos/farmacologia , Oviductos/efeitos dos fármacos , Útero/efeitos dos fármacos , Animais , Implantação do Embrião/efeitos dos fármacos , Desenvolvimento Embrionário/efeitos dos fármacos , Feminino , Humanos , Masculino , Ovulação/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos

RESUMO

OBJECTIVE: Ulispristal acetate (UPA) is a selective progesterone receptor modulator widely used for emergency contraception (EC). The described main mechanism of action is by inhibiting or delaying ovulation; however, the postovulatory effects of the drug are still on debate. Therefore, the aim of this study was to determine whether UPA could interfere with human sperm fertilizing ability. STUDY DESIGN: Human motile spermatozoa were incubated under capacitating conditions with or without UPA, and then used to inseminate human tubal explants, mouse cumulus-oocyte complexes and zona-free hamster eggs. The ability of UPA to interact with human sperm progesterone (P)-binding sites was investigated by incubating the cells with fluorescent-labeled P and analyzing them by fluorescence microscopy. RESULTS: UPA did not affect the ability of human sperm to bind to human tubal tissue explants surface or to penetrate the mouse cumulus mass and the zona-free hamster eggs. In addition, concentrations of UPA much higher than those present in the plasma of EC pill users were required to bind to human sperm P-binding sites. CONCLUSIONS: Our study supports a lack of an agonist or antagonist action of UPA on different functional parameters associated with the fertilizing ability of human sperm. IMPLICATIONS: This study provides new functional evidence supporting that the contraceptive action of UPA is not related to effects on human sperm cells, contributing to a better understanding of the mechanism of action of UPA as EC.

Assuntos

Anticoncepcionais Femininos/farmacologia , Tubas Uterinas/metabolismo , Norpregnadienos/farmacologia , Interações Espermatozoide-Óvulo/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Espermatozoides/fisiologia , Animais , Sítios de Ligação/efeitos dos fármacos , Anticoncepção Pós-Coito , Cricetinae , Células do Cúmulo/fisiologia , Feminino , Humanos , Masculino , Camundongos , Norpregnadienos/metabolismo , Progesterona/metabolismo , Receptores de Progesterona/efeitos dos fármacos

RESUMO

OBJECTIVE: Ulipristal acetate (UPA) acts as an emergency contraceptive by inhibiting ovulation. This study explores possible additional effects on the fragmentation of sperm DNA during in vitro incubation. METHODS: Motile spermatozoa from healthy donors were selected by swim-up and incubated under capacitating conditions in control medium or with UPA (1, 10, 100, 1,000 or 10,000 ng/ml). In some experiments, 200 µM of H2O2 were added to induce oxidative stress. The sperm chromatin dispersion test was performed to analyse DNA integrity (400 cells; 1000×). Lipid peroxidation (thiobarbituric acid assay), induced-acrosome reaction (AR) and sperm vitality (Eosin Y) were also evaluated in spermatozoa exposed to UPA and/or H2O2. RESULTS: During sperm incubation, the percentage of fragmented DNA increased significantly, from 15.0 ± 1.3 to 41.0 ± 4.5% (p < 0.001). In the presence of UPA, DNA fragmentation decreased significantly (p < 0.05), in a dose-dependent manner. At 100 and 1000 ng/ml, UPA also counteracted the effect of H2O2 and prevented DNA fragmentation. No effect on sperm vitality, lipid peroxidation or induced-AR was found with any treatment. CONCLUSIONS: During in vitro sperm capacitation DNA fragmentation increased but the latter was counteracted in the presence of UPA, which possibly acted as a scavenger of reactive oxygen species produced by spermatozoa.

Assuntos

Anticoncepcionais Sintéticos Pós-Coito/farmacologia , Fragmentação do DNA/efeitos dos fármacos , Norpregnadienos/farmacologia , Espermatozoides/efeitos dos fármacos , Reação Acrossômica/efeitos dos fármacos , Relação Dose-Resposta a Droga , Humanos , Peróxido de Hidrogênio/farmacologia , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Oxidantes/farmacologia , Estresse Oxidativo , Espermatozoides/fisiologia

RESUMO

OBJECTIVE: A pill containing ulipristal acetate (UPA) is used for emergency contraception (EC). Considering that, following its intake, spermatozoa may be exposed to UPA in the female genital tract we intended to evaluate sperm functions after incubation with this compound. METHODS: Motile spermatozoa were selected by swim-up and were incubated under capacitating conditions with UPA (at concentrations of 1, 10, 100, 1,000, and 10,000 ng/ml) or control medium. The main outcome measures were sperm vitality, sperm protein tyrosine phosphorylation (TyrP), spontaneous acrosomal reaction (AR), and human follicular fluid (hFF)-induced AR. RESULTS: Sperm vitality and TyrP pattern were similar between spermatozoa exposed to UPA or control. In addition, spontaneous AR ranged from 14.0 ±1.5% to 18.0 ±1.9% after exposure to UPA or control medium without significant differences, and UPA did not prevent hFF-induced AR. CONCLUSIONS: Incubation of sperm with UPA at concentrations around the expected plasma levels after ingestion of this EC pill (˜100-200 ng/ml) did not modify the signal transduction of TyrP involved in sperm capacitation. Moreover, UPA showed no agonist effect on progesterone receptors because it did not induce AR. Considering that progesterone in hFF is essential for AR induction, and UPA did not prevent the hFF-induced AR, an antagonist action of UPA on the AR is unlikely.

Assuntos

Reação Acrossômica/efeitos dos fármacos , Anticoncepcionais Sintéticos Pós-Coito/farmacologia , Norpregnadienos/farmacologia , Espermatozoides/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Feminino , Líquido Folicular/fisiologia , Humanos , Técnicas In Vitro , Masculino , Fosforilação/efeitos dos fármacos , Capacitação Espermática/efeitos dos fármacos , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/fisiologia

RESUMO

The aim of this study was to elucidate the mechanism involved in the acrosome reaction (AR) induced by follicular fluid (FF) in spermatozoa previously exposed to peritoneal fluid (PF). The influence of progesterone was also investigated. Semen samples were from 18 normozoospermic donors. PF samples were from 13 women with unexplained infertility and from a woman treated with synthetic progestagen. FF samples were collected from six women undergoing IVF/embryo transfer and pooled. Motile spermatozoa were capacitated overnight and a kinetic and inhibition study on the FF-induced AR was performed. Spermatozoa pretreated with PF were challenged with either FF or progesterone. The ability of progesterone- and progestagen-supplemented PF to induce AR was analysed. Enzyme-digested PF was also tested. Pre-incubation with PF for 60 min completely prevented the FF-induced AR; spermatozoa treated with PF were unable to respond to FF or progesterone and this effect was not reversible. Progesterone- and progestagen-supplemented PF stimulated the AR relative to controls. Enzyme-digested PF did not have an inhibitory capacity. These data strongly suggest that there are one or more inhibitory proteins in PF that interact with spermatozoa so as to prevent access of progesterone to its receptor and thus inhibit the occurrence of the AR. The oviduct, or Fallopian tube, provides a place for spermatozoa and egg transport and storage, fertilization and early embryo development. If ovulation has not occurred, spermatozoa may reside in the oviduct for several hours or even a few days, awaiting oocyte arrival. It is assumed that fluids present in the female genital tract may have a role in synchronizing the timing required to guarantee the success of fertilization. We previously observed that the peritoneal fluid that bathes the peritoneal cavity is a suitable medium for sperm survival and we also reported that this fluid could stabilize spermatozoa. In this study we show further evidence that the exposure to peritoneal fluid modifies the response of spermatozoa to oocyte signals.

Assuntos

Líquido Ascítico/fisiologia , Líquido Folicular/fisiologia , Espermatozoides/fisiologia , Reação Acrossômica/efeitos dos fármacos , Reação Acrossômica/fisiologia , Adulto , Líquido Ascítico/patologia , Regulação para Baixo , Feminino , Líquido Folicular/química , Líquido Folicular/metabolismo , Humanos , Técnicas In Vitro , Infertilidade Feminina/patologia , Cinética , Masculino , Progesterona/análise , Progesterona/metabolismo , Progesterona/farmacologia , Capacitação Espermática/efeitos dos fármacos , Capacitação Espermática/fisiologia , Motilidade dos Espermatozoides/efeitos dos fármacos , Motilidade dos Espermatozoides/fisiologia , Espermatozoides/efeitos dos fármacos , Adulto Jovem

RESUMO

OBJECTIVE: To determine the secretion of Grp78 by human oviduct epithelial cells, its association to spermatozoa, and its involvement in gamete interaction. DESIGN: Prospective study. SETTING: Basic research laboratory. SUBJECT(S): Semen samples obtained from normozoospermic volunteers. Tubal tissue provided by patients undergoing hysterectomies. Oocytes collected from women undergoing IVF-ET. INTERVENTION(S): Analysis of Grp78 expression and secretion by oviductal tissue. Gamete incubation with recombinant Grp78 (rec-Grp78). MAIN OUTCOME MEASURE(S): Assessment of protein expression and secretion by immunohistochemistry and Western immunoblotting, respectively. Evaluation of rec-Grp78 binding to human spermatozoa by immunocytochemistry, and analysis of its effect upon gamete interaction using the hemizona assay. RESULT(S): Grp78 was found in the surface of oviduct epithelial cells. Soluble Grp78 was detected in oviductal fluids from women in the periovulatory period and in oviductal tissue conditioned medium. Rec-Grp78 was able to bind to the sperm acrosomal cap, and its presence during gamete interaction led to a decrease in the number of spermatozoa bound to the zona pellucida (ZP). When calcium ions from the incubation medium were replaced by strontium, rec-Grp78 enhanced sperm-ZP interaction. CONCLUSION(S): Grp78 is expressed and secreted by oviduct epithelial cells. The protein would bind to the gametes and may modulate their interaction in a calcium-dependent manner.

Assuntos

Células Epiteliais/metabolismo , Tubas Uterinas/metabolismo , Proteínas de Choque Térmico/metabolismo , Interações Espermatozoide-Óvulo , Espermatozoides/metabolismo , Zona Pelúcida/metabolismo , Adulto , Western Blotting , Cálcio/metabolismo , Linhagem Celular Tumoral , Células Cultivadas , Chaperona BiP do Retículo Endoplasmático , Feminino , Humanos , Imuno-Histoquímica , Masculino , Ciclo Menstrual , Pessoa de Meia-Idade , Ligação Proteica , Proteínas Recombinantes/metabolismo , Técnicas de Cultura de Tecidos

RESUMO

OBJECTIVE: To study the effect of the human tubal tissue conditioned medium (CM) on sperm parameters related to sperm-zona pellucida interaction. DESIGN: Controlled experimental laboratory study. SETTING: Research laboratory. SUBJECT(S): Semen samples from donors with normozoospermia. Human tubal tissue obtained from women undergoing hysterectomies. Human follicular fluids (hFF) and oocytes collected from patients undergoing IVF-ET. INTERVENTION(S): Incubation of spermatozoa with CM proteins obtained from human tubal tissue culture; sperm binding to the zona pellucida assessment. MAIN OUTCOME MEASURE(S): Explants' viability was assessed by tissue DNA analysis. Sperm ability to interact with zona was tested with use of the whole oocyte test. Expression of d-mannose binding sites was assessed with use of a fluorescent probe on mannose coupled to bovine serum albumin. Human FF-induced acrosome reaction was assessed by the Pisum sativum technique. RESULT(S): Although treatment with 0.8 microg/microL of CM allowed sperm binding to the zona and the expression of d-mannose binding sites comparable with sperm in control medium, with 3.2 microg/mL of CM resulted in a significant decrease of both parameters. No effect of CM on spontaneous or hFF-induced acrosome reaction or in sperm viability was observed. CONCLUSION(S): The results indicate that the incubation of spermatozoa in the presence of CM reduces sperm affinity for the zona pellucida. This effect can be partly explained by the decreased expression of d-mannose binding sites on the sperm surface.

Assuntos

Reação Acrossômica , Tubas Uterinas/metabolismo , Espermatozoides/metabolismo , Zona Pelúcida/metabolismo , Adulto , Sítios de Ligação , Sobrevivência Celular , Meios de Cultivo Condicionados/metabolismo , Dano ao DNA , Feminino , Líquido Folicular/metabolismo , Humanos , Masculino , Manose/metabolismo , Pessoa de Meia-Idade , Técnicas de Cultura de Tecidos

RESUMO

PURPOSE: In the female genital tract spermatozoa interact with the oviductal secretion. The aim of the study was to evaluate the effect of conditioned media (CM) from cultures of human oviductal tissue, on sperm DNA integrity. The effect of H(2)O(2) on sperm DNA integrity, before and after incubation under capacitating conditions, was also investigated. METHODS: Motile sperm obtained from normozoospermic semen samples were incubated (4 h or 22 h) in the presence or absence of CM and further exposed to H(2)O(2). DNA damage was detected by the comet assay. RESULTS: The CM significantly reduced the DNA damage associated with sperm incubation, and also decreased the effect of H(2)O(2) after 4 h incubation, compared to controls. The H(2)O(2) caused a dose-dependent deleterious effect on sperm DNA integrity both before and following 22 h of capacitation. CONCLUSION: The oviductal tissue CM increased the stabilization of the sperm DNA structure under culture conditions.

Assuntos

Líquidos Corporais/fisiologia , Dano ao DNA , Tubas Uterinas/metabolismo , Espermatozoides/metabolismo , Adulto , Algoritmos , Líquidos Corporais/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Ensaio Cometa , DNA/efeitos dos fármacos , DNA/metabolismo , DNA/fisiologia , Dano ao DNA/fisiologia , Feminino , Humanos , Peróxido de Hidrogênio/farmacologia , Masculino , Pessoa de Meia-Idade , Capacitação Espermática/efeitos dos fármacos , Capacitação Espermática/fisiologia , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Espermatozoides/fisiologia

RESUMO

The aim of the present study was to further evaluate the participation of D-mannose in the process of human sperm-egg interaction. Zona pellucida binding competitive assays in the presence of D-mannose were carried out using discarded oocytes from IVF. Spermatozoa were capacitated and D-mannose-binding site (MBS) expression, sperm viability and follicular fluidinduced acrosome reaction (AR) were evaluated. MBS were visualized using a fluorescein-neoglycoprotein probe. The capacity of free D-mannose and mannosylated albumin to induce the AR was also tested. MBS and the IVF outcome were also analysed. The involvement of D-mannose in sperm binding to the zona pellucida was verified by the inhibitory effect produced when the sugar was present during binding assays. MBS expression increased during capacitation, in parallel with the ability to undergo the induced AR. Mannosylated albumin, but not the free sugar, induced the AR. In acrosome-reacted spermatozoa, the MBS was located at the plasma membrane, as shown by confocal analysis. No significant difference in the increase in MBS expression was observed among the different IVF groups of patients. The data show that D-mannose is involved in the sperm-zona pellucida interaction, and that the expression of MBS on the sperm surface occurs during the acquisition of in-vitro sperm fertilizing ability.

Assuntos

Manose/metabolismo , Interações Espermatozoide-Óvulo/fisiologia , Espermatozoides/metabolismo , Zona Pelúcida/metabolismo , Adulto , Ligação Competitiva , Feminino , Fertilização in vitro , Humanos , Masculino , Oócitos/metabolismo

RESUMO

Despite the fact that both peritoneal (PF) and follicular (FF) fluids have a common ovarian origin, FF is a natural inducer of sperm acrosome reaction (AR) while PF is not. To better understand these effects, concentrations of oestradiol, progesterone and proteins in peri-ovulatory PF and FF were determined and compared. PF was aspirated by laparoscopy at the peri-ovulatory stage from women with unexplained infertility. FF was collected from patients undergoing IVF and pooled. PF and FF were tested for the presence of antisperm antibodies. Oestradiol and progesterone were measured by enzyme immunoassay, and total protein concentration was determined and analysed. The AR was determined in spermatozoa that were exposed to PF alone, progesterone-supplemented PF, progesterone, control medium, or ethanol. No antisperm antibodies were found in any fluid tested. Oestradiol and progesterone and concentrations in PF were significantly lower than in FF. Protein concentration was also significantly lower in PF than in FF, but no differences were observed between the electrophoretic patterns. When capacitated spermatozoa were exposed to progesterone-supplemented PF there was a significant increase in the percentage of AR with respect to those in PF, control medium or ethanol. These results suggest that the lack of AR-stimulating activity of PF was related to its lower progesterone concentration compared with FF.

Assuntos

Reação Acrossômica/fisiologia , Líquido Ascítico/química , Estrogênios/análise , Líquido Folicular/química , Progesterona/análise , Espermatozoides/fisiologia , Adulto , Anticorpos/análise , Feminino , Humanos , Masculino , Progesterona/farmacologia , Proteínas/análise , Espermatozoides/efeitos dos fármacos , Espermatozoides/imunologia

RESUMO

It has been suggested that oviductal proteins could be involved in modulating sperm function and fertilizing ability through as yet not well-known mechanisms. The objective of the study was to investigate the pattern of proteins secreted by human oviductal tissue cultures and the effects of their conditioned media (CM) on sperm function under capacitating conditions and in phosphate buffered saline (PBS). In addition, interactions between spermatozoa and oviductal proteins were examined. The oviductal tissue was obtained from pre-menopausal patients scheduled for hysterectomies because of uterine fibromyoma. Normozoospermic semen samples were obtained from healthy donors. Cultures of human fallopian tissue were carried out and CM were collected for analysis of the de novo production of [35S]-methionine-labelled proteins by SDS-PAGE. Motile spermatozoa were incubated under capacitating conditions and in PBS, with or without CM, and sperm fertilizing ability was assessed by ionophore-induced acrosome reaction (AR) and the acrosome reaction to ionophore challenge (ARIC) score. The ionophore-induced AR was evaluated by the Pisum sativum technique. Sixteen de novo produced proteins were detected in CM. One of these proteins (molecular weight 79 kDa) was detected in extracts from spermatozoa pre-incubated with CM. Sperm survival and motility were maintained in the presence of CM, although results showed a significant decrease in ARIC score (p < 0.05), with respect to controls. The presence of CM significantly decreased sperm fertilizing ability, without affecting sperm survival. These results suggest that the oviductal secretion could contribute to preserve sperm viability and motility, and to prevent a premature response of spermatozoa to AR inducers.

Assuntos

Tubas Uterinas/citologia , Tubas Uterinas/metabolismo , Motilidade dos Espermatozoides/fisiologia , Espermatozoides/fisiologia , Sobrevivência Celular , Células Cultivadas , Eletroforese em Gel de Poliacrilamida , Feminino , Humanos , Masculino , Metionina/metabolismo , Proteínas/isolamento & purificação , Proteínas/metabolismo , Sêmen/fisiologia , Espermatozoides/citologia

RESUMO

OBJECTIVE: To examine the effect of peritoneal fluid on various parameters of sperm function in vitro. DESIGN: Prospective study. SETTING: Basic research laboratory. PATIENT(S): Semen samples were obtained from normozoospermic volunteers (n = 43). Peritoneal fluids were aspirated laparoscopically from women with unexplained infertility (n = 14). Follicular fluid and oocytes were collected from patients undergoing IVF-ET. INTERVENTION(S): Sperm incubated under capacitating conditions were exposed to peritoneal fluid, and functional variables were evaluated in vitro. MAIN OUTCOME MEASURE(S): Sperm viability and motility, follicular fluid and calcium ionophore-induced acrosome reactions, protein tyrosine phosphorylation, expression of D-mannose binding sites, and ability of sperm to interact with zona pellucida. RESULT(S): Exposure of sperm to peritoneal fluid for up to 6 hours did not affect sperm viability or motility. Unlike follicular fluid, peritoneal fluid did not induce the acrosome reaction. Moreover, incubation of sperm with > or =20% v/v peritoneal fluid for 1 hour prevented the follicular fluid and the ionophore-induced acrosome reaction. Although treatment with peritoneal fluid allowed protein tyrosine phosphorylation during capacitation, it resulted in a significant decrease in the expression of D-mannose binding sites and sperm-zona pellucida binding. CONCLUSION(S): Peritoneal fluid maintains sperm survival and decreases sperm ability to respond to inducers of the acrosome reaction and bind to the zona pellucida in vitro, indicating that this fluid might modulate sperm function in vivo.

Assuntos

Líquido Ascítico/metabolismo , Infertilidade/fisiopatologia , Espermatozoides/fisiologia , Reação Acrossômica , Adulto , Sítios de Ligação , Sobrevivência Celular , Feminino , Líquido Folicular/metabolismo , Humanos , Masculino , Manose/metabolismo , Fosforilação , Estudos Prospectivos , Capacitação Espermática , Motilidade dos Espermatozoides , Interações Espermatozoide-Óvulo , Espermatozoides/metabolismo , Tirosina/metabolismo

Assuntos

Humanos , Masculino , Adulto , Capacitação Espermática/efeitos dos fármacos , Oxidantes/biossíntese , Reação Acrossômica/efeitos dos fármacos , Espermatozoides/metabolismo , Oxidantes/efeitos adversos , Espécies Reativas de Oxigênio , Sêmen/química , Fertilização in vitro/efeitos dos fármacos

Assuntos

Humanos , Masculino , Adulto , Capacitação Espermática , Oxidantes/biossíntese , Reação Acrossômica , Espécies Reativas de Oxigênio , Fertilização in vitro , Oxidantes/efeitos adversos , Sêmen/química , Espermatozoides/metabolismo

RESUMO

El objetivo de éste trabajo fue determinar mediante un modelo in vitro, el efecto que ejerce el fluido peritoneal sobre la función del espermatozoide humano, tomando como control al fluido folicular. Para ello se obtuvieron fluidos peritoneales provenientes de pacientes que realizaron laparascopia diagnóstica en fecha periovulatoria y que no presentaron enfermedad tubo-peritoneal (n=9). El fluido folicular se obtuvo de pacientes que realizaron recuperación ovocitaria en un programa de fertilización in vitro (n=4). Ambos fluidos fueron centrifugados, filtrados y congelados a -20ºC. Los sémenes de pacientes normozoospérmicos (n=21) fueron procesados mediante un gradiente discontínuo de Percoll y capacitados en medio sintético Human Tubal Fluid suplementado con albúmina, a una concentración de 2-10 por 10 esp./ml durante 3 a 4 hora

Assuntos

Humanos , Masculino , Feminino , Técnicas In Vitro , Interações Espermatozoide-Óvulo/fisiologia , Peritônio/metabolismo , Líquido Folicular , Reação Acrossômica , Zona Pelúcida , Manose/diagnóstico , Ionóforos/diagnóstico , Capacitação Espermática

RESUMO

El objetivo de éste trabajo fue determinar mediante un modelo in vitro, el efecto que ejerce el fluido peritoneal sobre la función del espermatozoide humano, tomando como control al fluido folicular. Para ello se obtuvieron fluidos peritoneales provenientes de pacientes que realizaron laparascopia diagnóstica en fecha periovulatoria y que no presentaron enfermedad tubo-peritoneal (n=9). El fluido folicular se obtuvo de pacientes que realizaron recuperación ovocitaria en un programa de fertilización in vitro (n=4). Ambos fluidos fueron centrifugados, filtrados y congelados a -20§C. Los sémenes de pacientes normozoospérmicos (n=21) fueron procesados mediante un gradiente discontínuo de Percoll y capacitados en medio sintético Human Tubal Fluid suplementado con albúmina, a una concentración de 2-10 por 10 esp./ml durante 3 a 4 horas. En experimentos apareados, los espermatozoides capacitados fueron expuestos en relación 1:1 v/v al medio control, al fluido folicular o al peritoneal durante 1 hora, luego de lo cual se procedió a estudiar la función espermática mediante los siguientes tests: a) viabilidad b) motilidad progresiva c) la reacción acrosomal inducida por ionóforo de calcio y d) la expresión del receptor espermático para D-manosa. Nuestros resultados muestran que ni la viabilidad ni la motilidad resultaron afectadas por el pre-tratamiento. Si bien la ocurrencia de la reacción acrosomal espontánea no fue diferente entre los distintos grupos (6.3 a 7.5 por ciento, NS), la inducida por ionóforo disminuyó significativamente luego de la exposición al fluido peritoneal y al folicular (18.9ñ5.1 por ciento y 29.0ñ6.1 por ciento vs control 45.6ñ5.0 por ciento, p <0.001 y 0.01 respectivamente). Si bien la detección del receptor para D-manosa aumentó durante la capacitación, resultó significativamente menor a los controles luego del tratamiento previo (FP: 15.4ñ2.9 por ciento y FF: 15.5ñ3.1 por ciento vs control 23.7ñ3.1 por ciento, p<0.05. Nuestros resultados sugieren que al menos in vitro, tanto el fluido peritoneal como el folicular modularían la función del espermatozoide humano manteniéndolo vivo y móvil, pero en un estado no reaccionado

Assuntos

Humanos , Masculino , Feminino , Técnicas In Vitro , Interações Espermatozoide-Óvulo/fisiologia , Reação Acrossômica , Líquido Folicular , Ionóforos , Manose , Peritônio/metabolismo , Capacitação Espermática , Zona Pelúcida