In vitro assessment of reproductive toxicity of cigarette smoke and deleterious consequences of maternal exposure to its constituents

Cigarette smoke is known to be a serious health risk factor and considered reproductively toxic. In the current study, we investigated whether constituents of cigarette smoke, pyrazine, 2-ethylpyridine, and 3-ethylpyridine, adversely affect reproductive functioning such as oocyte maturation and sperm capacitation. Our findings indicated that three smoke components were involved in retardation of oocyte maturation in a dose-dependent manner and the lowest-observed-adverse-effect level (LOAEL) was determined to be 10-10M. However, individual smoke components administrated at the LOAEL did not attenuate oocyte maturation, demonstrating that all three toxicants were equally required for the observed growth impairment. When exposed to all three components at 10-10M during in vitro capacitation, murine sperm lost forward progression and were unable to show adequate hyperactivation, which is indicative of the incompletion of the capacitation process. Only sperm administrated with 3-ethylpyridine alone showed significant reduction in capacitation status, suggesting the chemical is the one responsible for disrupting sperm capacitation. Taken together, this is the first report that documents the effect of cigarette smoke components on oocyte maturation and sperm capacitation. The present findings demonstrate the adverse effects of smoke constituents of mammalian reproduction and the differences in sensitivity to smoke components between male and female gametes. Since both processes take place in the female reproductive system, our data provide new insights into deleterious consequences of maternal exposure to cigarette smoke.

Porcine oviduct sperm binding glycoprotein and its deleterious effect on sperm: a mechanism for negative selection of sperm?

In their journey through the oviduct some subpopulations of sperm are preserved in a reservoir, while others are negatively selected. Sperm binding glycoprotein (SBG) is a pig oviductal epithelial cell glycoprotein that produces, under capacitating conditions, acrosome alteration, p97 tyrosine-phosphorylation and reduction of the motility of sperm. In this paper, we show that SBG is accessible at the extracellular surface of the oviductal epithelial cells, supporting a sperm interaction biological role in situ. We analyze the possible dependence of the tyrosine-phosphorylation of p97 on the PKA mechanism, finding that apparently it is not PKA dependent. Also, after SBG treatment the phosphorylated proteins locate mainly at the detached periacrosomal region and at the tail of sperm; the latter may be related to SBG's motility reduction effect. The study of the time course effect of SBG on sperm as detected by chlortetracycline (CTC) staining and of its binding to sperm by immunodetection in conjunction with CTC, shows results in agreement with the hypothesis that this glycoprotein is involved in the alteration of acrosomes in a specific sperm subpopulation. The results suggest that SBG may be part of a mechanism for negative selection of sperm.

Acrosin release and acrosin activity during incubation in capacitating media using fresh and frozen-thawed dog sperm

We evaluated the effect of time and temperature on acrosin release from the acrosomal cap and the activity of this enzyme during in vitro capacitation in fresh and frozen/thawed dog sperm. Sperm-rich fractions of six ejaculates from three dogs were processed as fresh and frozen samples. Each sperm sample was incubated in canine capacitation medium (CCM) for 0, 1, 2 and 3 h at 20°C and at 37°C. After incubation, the samples were assessed by the indirect immunofluorescent staining technique. The probability of having unlabeled sperm (PUS), indicating acrosin loss, was modelled by a binomial distribution using logistic regression. There was a linear relationship between PUS and time at both temperatures (p<0.001); however, a major percentage of unlabeled sperm was observed in frozen/thawed samples soon after incubation, indicating that the release of acrosin was affected by capacitation time, mainly in frozen samples. Temperature influenced acrosin release only in cryopreserved sperm (p<0.05). Acrosin activity was measured by digestion halos on slides coated with gelatin-substrate film during each time period; a significant increase in the number of large halos was observed in fresh samples throughout the experiment, whereas frozen/thawed sperm showed a decreased rate of halo diameters during culture. Thus, there appears to differences between fresh and frozen dog sperm in terms of acrosin release and the level of acrosin activity in the course of in vitro capacitation.

Modulation of spermatozoon acrosome reaction

Spermatozoon acrosome reaction is an exocytotic event of the utmost importance for the development of mammalian fertilisation. Current evidence shows that the triggering of the acrosome reaction (AR) could be regulated by the action of diverse compounds, namely, metabolites, neurotransmitters and hormones. The aim of the present review is to describe the modulating effects of several compounds that have been classified as inductors or inhibitors of acrosome reaction. Among AR inductors, it is necessary to mention progesterone, angiotensin II, atrial natriuretic peptide, cathecolamines, insulin, leptin, relaxin and other hormones. Regarding the inhibitors, oestradiol and epidermal growth factor are among the substances that retard AR. It is worth mentioning that gamma-aminobutyric acid, a neurotransmitter known to be an inhibitor in the central nervous system, has been shown to induce AR. The multiple hormones located in the fluids of the female reproductive tract are also likely to act as subtle regulators of AR, constituting a fundamental aspect for the development of successful fertilisation. Finally, it is necessary to emphasise that the study of regulation exerted by hormones and other compounds on AR is essential for further understanding of mammalian reproductive biology, especially spermatozoon physiology.

Características andrológicas e do sêmen de touros do composto Red Norte (Nelore x Tabapuã x Red Angus x Sinepol) / Andrological and semen characteristics of cross-bred Red Norte (Nelore x Tabapuã x Red Angus x Sinepol) young bulls

Avaliaram-se as características andrológicas do sêmen de touros jovens do composto Red Norte (Nelore x Tabapuã x Red Angus x Sinepol), com idade média de 13,9±0,8 meses, com o objetivo de estimar o advento da puberdade e a qualidade do sêmen. Foram avaliados o perímetro escrotal (PE), o peso e as características seminais de 70 tourinhos, classificados em três grupos, de acordo com o PE: GI=27-33cm (n=24), GII=33-35cm (n=24) e GIII=35-43cm (n=22). As médias de peso e a idade de cada grupo (G) foram, respectivamente: GI=411,2±37,4kg e 13,8±1,0 meses, GII=426,9±31,5kg e 14,0±0,7 meses e GIII=438,4±38,3kg e 14,0±0,6 meses. As características seminais para cada grupo foram, volume 4,2±3,1mL, 5,3±2,6mL e 4,5±2,1mL; motilidade 31,3±24,1 por cento, 44,2±23,9 por cento e 43,9±21,5 por cento e vigor 2,8±1,6, 3,5±1,3 e 3,5±1,3, respectivamente. O espermiograma apresentou valores médios de concentração de 130,5±266,2x10(6)/mL, 289,5±390,2x10(6)/mL e 333,9±523,7x10(6)/mL, defeitos totais de 81,4±15,9 por cento, 73,8±15,4 por cento e 67,9±19,0 por cento; defeitos maiores de 87,3±26,2 por cento, 66,8±24,9 por cento e 56,7±17,1 por cento e defeitos menores de 16,6±14,9 por cento, 33,2±24,9 por cento e 43,3±17,1 por cento, respectivamente. Dos setenta animais examinados, sete (10 por cento) foram considerados aptos à reprodução. Os resultados mostraram que a patologia espermática diminuiu em razão do aumento do PE.

Reproductive traits of cross-breed Red Norte (Nelore x Tabapuã x Red Angus x Sinepol) young bulls averaging of 13.9±0.8 month-old were evaluated, in order to determine the puberty onset and semen quality in these animals. Scrotal circumference (SC), body weight (BW), and semen parameters of 70 bulls were measured. Animals were allotted in three groups (G) according to their SC: GI=27-33cm (n=24), GII=33-35cm (n=24), and GIII=35-43cm (n=22). BW and age of each group were, respectively: GI=411.2±37.4kg and 13.8±1.0 month-old, GII=426.9±31.5kg and 14.0±0.7 month-old, and GIII=438.4±38.3kg and 14.0±0.6 month-old. Seminal physical characteristics for same order of groups were: volume 4.2±3.1mL, 5.3±2.6mL, and 4.5±2.1mL; motility 31.3±24.1 percent, 44.2±23.9 percent, and 43.9±21.5 percent; and vigor 2.8±1.6, 3.5±1.3, and 3.5±1.3. The spermiogram presented concentration values of 130.5±266.2x10(6)/mL, 289.5±390.2x10(6)/mL, and 333.9±523.7x10(6)/mL; total defects of 81.4±15.9 percent, 73.8±15.4 percent, and 67.9±19.0 percent; major defects of 87.3±26.2 percent, 66.8±24.9 percent and 56.7±17.1 percent; and minor defects of 16.6±14.9 percent, 33.2±24.9 percent, and 43.3±17.1 percent, for same order of groups. Seven out of 70 bulls were considered satisfactory potential breeders. Results showed that semen pathology progressively decreased when SC increased.

Binding characteristics of sperm with recombinant human zona pellucida glycoprotein-3 coated beads.

Background & objectives: An inability or decreased ability of spermatozoa to bind to the zona pellucida (ZP), an extracellular glycoproteinaceous matrix surrounding egg, is one of the plausible causes of idiopathic infertility. It will be clinically useful to distinguish this condition from other causes of infertility. An assay system, investigating binding of human sperm with ZP glycoprotein may prove useful in this regard. We attempted to develop a simple assay system to analyse the binding of capacitated human spermatozoa to human zona pellucida glycoprotein-3 (ZP3) using baculovirus-expressed recombinant human ZP3 coated beads. Methods: Recombinant baculovirus-expressed ZP3 was purified, labelled with biotin and coated on streptavidin sepharose beads. An in vitro assay system was optimized to study binding of capacitated human sperm to ZP3 coated beads. Results: A higher percentage of baculovirus-expressed recombinant human ZP3 coated beads showed significant (P<0.05) binding of capacitated human sperm as compared to beads coated with fetuin. An inhibition in the binding of sperm to ZP3 coated beads was observed in presence of cold recombinant human ZP3. Further, prior incubation of ZP3 coated beads with monoclonal antibodies (MAbs) against ZP3 but not against ZP2 resulted in the decrease in number of sperm bound to bead. Interpretation & conclusion: An in vitro assay system to study the binding of human sperm to ZP3- primary sperm receptor was established, which may be useful to determine the functional competence of spermatozoa.

Frozen Jaguar (Panthera onca) sperm capacitation and ability to penetrate zona free hamster oocytes / Capacitação espermática de sêmem congelado de onça pintada (Panthera onca) e ensaio de penetração em oócitos de hamster livres de zona pelúcida

Assisted reproductive technologies in endangered species, such as artificial insemination, in vitro fertilization and embryo transfer, can be viewed as one potential approach for safeguarding species. Toward this aim, the objective of this study was to evaluate the fertility of frozen jaguar (Panthera onca) sperm and Tyrod's Talp PVA capacitation medium using the hamster zona free oocyte penetration assay. Ejaculates were collected from nine animals using electroejaculation and cryopreserved. Sperm capacitation was performed by swim-up technique using Tyrod's Talp PVA medium at room temperature. Penetration was considered when the spermatozoa head decondensation was visualized within the oocyte. This assay showed 15.4 % penetrations (350/2275 oocytes). Results of this study showed high sperm abnormalities, low sperm quality after cryopreservation, and low percentage of penetrations. However, the penetration results showed lhat the cryopreserved jaguar's semen can be used for artificial insemination, in vitro fertilization and intra cytoplasmatic sperm injection, supporting the semen bank creation for this specie.

Biotecnologias reprodutivas aplicadas a espécies selvagens, como inseminação artificial, fertilização in vitro e transferência de embriões, são vistas como um potencial caminho para proteção das espécies ameaçadas de extinção. Devido a isso, o objetivo deste estudo foi avaliar a fertilidade do sêmen congelado de onças pintadas (Panthera onca) e o meio de capacitação espermática usando o ensaio de penetração em oócitos de hamster livres de zona pelúcida. Ejaculados de nove animais foram coletados por eletroejaculação e criopreservados. Para determinar a capacitação espermática foi utilizada a técnica swin-up com meio Tyrods Talp PVA a temperatura ambiente. No ensaio de penetração em oócitos de hamster livres de zona pelúcida foram considerados como oócitos penetrados aqueles que apresentaram em seu interior a cabeça do espermatozóide descondensada. O ensaio foi realizado em um total de 2275 oócitos, dos quais 350 apresentaram em seu interior a cabeça do espermatozóide descondensada, perfazendo um total de 15.4% de penetração. Os resultados deste estudo demonstraram alto índice de anormalidades espermáticas, baixa qualidade do sêmen e baixa porcentagem de penetrações. Entretanto, os resultados de penetração espermática demonstraram que sêmen congelado de onça pintada poderá ser utilizado para inseminação artificial, fertilização in vitro e injeção intracitoplasmática dando suporte para a criação de um banco de sêmen para esta espécie.

Avaliação histomorfométrica do parênquima testicular de ratos adultos tratados com diferentes doses de ivermectina / Histomorphometric evaluation of testicular parenchyma of adult rats treated with different dosages of ivermectin

Avaliou-se o efeito da ivermectina sobre o parênquima testicular através da produção espermática diária e da eficiência da espermatogênese em ratos Wistar adultos tratados com diferentes dosagens (200, 400 e 600æg/kg). Pela avaliação histomorfométrica, o parênquima testicular e o processo espermatogênico dos ratos Wistar não sofreram qualquer efeito deletério da aplicação de ivermectina, o que foi confirmado pela manutenção da produção espermática diária por testículo, pelo rendimento intrínseco da espermatogênese (PED/g/t) e pela manutenção da estrutura do parênquima testicular. Com base nos resultados quantitativos e qualitativos da espermatogênese, é possível concluir que a ivermectina não tem efeito tóxico-degenerativo sobre o parênquima testicular de ratos Wistar adultos.

The aim of this work was to evaluate the ivermectin effect on the testicular parenchyma through the daily spermatic production and the efficiency of the spermatogenesis in adult Wistar rats treated with different dosages (200, 400 and 600æg/kg) of ivermectin. Based on the histomorfometric evaluation, ivermectin had no deleterious effect on the testicular parenchyma and spermatogesis, which one was confirmed through the maintenance of the daily spermatic input and intrinsic income of spermatogenesis (PED/g/t), as well as by the maintenance of the testicular parenchyma structure. Based on the quantitative and qualitative results of spermatogenesis, it is possible to conclude that ivermectin does not have toxic-degenerative effect on the testicular parenchyma of adult Wistar rats.

Proacrosin-acrosin activity in capacitated and acrosome reacted sperm from cryopreserved bovine semen

Acrosin activity is associated with normal fertility in human and bovine spermatozoa. The aim of the study was to determine the variation of the enzyme activity in the proacrosin-acrosin system in capacitated and acrosome recated cryopreserved bovine sperm. Enzyme activity was assessed spectrophotometrically using N-alpha-benzoyl-DL-arginine p-nitroanilide (BAPNA) as specific substrate for acrosin at pH 8. Capacitation with heparin and quercitin failed to induce conversion of proacrosin to acrosin. An increase in acrosin activity produced by the presence of progesterone, in a dose-dependent manner, was related with the induction of true acrosome reaction. The total level of acrosin activity registered showed that 96 per cent of acrosin of capacitated sperm samples and control is present in the zymogen form. Moreover, progesterone is capable of duplicating the level of active enzyme, indicating that enzyme activity changes are related to acrosome reaction, suggesting that only a minor proportion of the total of proacrosin-acrosin system is required in the exocytotic process on cryopreserved bovine sperm.

Lipid peroxidation and antioxidant enzymes in male infertility.

BACKGROUND AND AIM: Mammalian spermatozoa are rich in polyunsaturated fatty acids and are very susceptible to attack by reactive oxygen species (ROS) and membrane lipid peroxide ion. Normally a balance is maintained between the amount of ROS produced and that scavenged. Cellular damage arises when this equilibrium is disturbed. A shift in the levels of ROS towards pro-oxidants in semen and vaginal secretions can induce an oxidative stress on spermatozoa. The aim was to study lipid peroxidation and antioxidant enzymes such as catalase, glutathione peroxidase and superoxide dismutase (SOD) and to correlate the same, with the 'water test', in male infertility. SETTINGS: Experimental study. SUBJECTS AND METHODS: Ejaculates from a total of 83 infertile and fertile healthy individuals were obtained. Lipid peroxidation and antioxidant enzyme levels were studied and correlated with water test. RESULTS: The results indicate that (i) the antioxidant enzyme catalase showed no significant changes in the various pathological samples, (ii) antioxidant enzymes SOD and glutathione peroxidase correlate positively with asthenozoospermic samples and (iii) the degree of lipid peroxidation also correlates positively with the poorly swollen sperm tails. The increase in SOD and glutathione peroxidase values, in the pathological cases represents an attempt made to overcome the reactive oxygen species. CONCLUSION: Water test could be used as a preliminary marker test for sperm tail damage by reactive oxygen species, since it correlates very well with lipid peroxidation and antioxidant enzymes.

El diagnóstico y tratamiento del factor masculino en la era del ICSI / Diagnosis and treatment of male factor in ICSI times

Actualmente somos testigos del avance en las técnicas de Reproducción Asistida que con el advenimiento de la inyección intracitoplasmática de esperma (ICSI), han venido a cumplir los objetivos reproductivos en pacientes que cuentan con un factor masculino de base. Sin embargo, el uso de estas técnicas de micromanipulación ha dado una pauta reproductiva, en ausencia de un diagnóstico etiológico y fisiopatológico. El estudio andrológico sistematizado de las características dinámicas y los aspectos funcionales espermáticos, complementados por la valoración endocrinourológica deben implementarse para un manejo diagnóstico y terapéutico reproductivo adecuado.

Surface protein changes in goat spermatozoa during capacitation.

Polypeptides of goat sperm surface before and after capacitation were examined by radiolabelling and immunologically using polyclonal antisera. Radioiodination revealed five protein bands having mol wt of 14.8, 72.4, 81, 100 and 128 kDa in uncapacitated ejaculated spermatozoa and only three bands of 23.4, 27 and 72.4 KDa in capacitated spermatozoa. The protein band with mol wt 72.4 kDa was only feebly iodinated in uncapacitated sperm surface but in capacitated spermatozoa it was heavily labelled. Western blot analysis of detergent-extracted proteins using gamma-globulin fraction of antisera raised against purified goat sperm plasma membrane revealed six antigens (17.8, 29.1, 33.4, 45.6, 85.1, 123.2 kDa) in uncapacitated spermatozoa, four (26, 32.1, 40.1, 45.6 kDa) in capacitated spermatozoa and only one (45.6 kDa) in acrosome-reacted spermatozoa. High mol wt proteins were more numerous on the surface of uncapacitated spermatozoa while the capacitated spermatozoa had relatively low mol wt proteins. An apparent effect of capacitation is the metabolism and reorganisation of proteins on goat sperm surface. Polypeptides on capacitated sperm surface revealed through radiolabelling and polyclonal antisera may have a likely receptor(s) role in the recognition and binding to homologous zona pellucida during fertilization.

Capacitaçäo espermática em ruminantes / Sperm capacitation in ruminants

A autora apresenta uma revisão de literatura da capacitação dos espermatozóides de ruminantes, relatando os prováveis mecanismos fisiológicos que estão envolvidos no processo e os agentes que provocam a capacitação in vitro dos espermatozóides.

Role of estradiol in the capacitation and acrosome reaction of hamster epididymal spermatozoa in the isolated uterus of mice incubated in vitro.

The site of sperm capacitation, the agents and mechanisms causing capacitation and acrosome reaction (AR) in vivo are not well understood. The female reproductive tract has been reported to play a key role during capacitation and AR. Some experiments were carried out on the capacitation and AR of hamster epididymal spermatozoa in the estrogen and progesterone dominated uterus (estrous and diestrous respectively) albino mice, incubated in TALP without calcium and BSA. Also the effect of estradiol (200 micrograms/ml) supplemented to TALP, on capacitation and AR was examined. Capacitation and AR of hamster spermatozoa incubated in the isolated uterus of both estrous and diestrous mice were significantly (P < 0.05) higher in the presence of exogenous estradiol than that in its absence. Acrosome shedding occurred earlier i.e. at 3rd hour as compared to the in vitro studies where it occurred at 5th hour. The present study thus reveals that uterus of both estrogen and progesterone dominated mice play an important role in the induction of capacitation and AR. The addition of estradiol might have the influence on the synthesis of uterine proteins of mice which might be important for capacitation and AR.

Proacrosin and acrosin determination during capacitation and acrosome reaction in rabbit spermatozoa

The proacrosin/acrosin system has been immunolocalized, by the silver enhanced immunogold technique on the acrosomal region of capacitated and perivitelline rabbit spermatozoa. The purpose of the present work was to investigate the kinetics of the activation of acrosin by determining the proportion of proacrosin and acrosin present in the acrosome of the rabbit spermatozoa during capacitation and the induction of the acrosome reaction by the calcium ionophore A23187. Rabbit spermatozoa selected by the percoll gradient technique were incubated for 0, 0.5 and 6 hours and then the acrosome reaction was induced at 0 and 6 hours with 1.9 microM of the calcium ionophore A23187. It was found that 95 of acrosin activity in rabbit spermatozoa at zero time corresponds to proacrosin and after capacitation and acrosome reaction, a diminution of the activity of proacrosin/acrosin system was found. However, proacrosin represents the large majority of the system activity. Western blot prepared with sperm extract obtained at the start of incubation showed the characteristic doublet band about 53-55 kDa that may correspond to proacrosin and alpha-acrosin. After six hours of incubation and with induction with the calcium ionophore A23187 the same doublet was seen in addition to a third band of 49 kDa that could correspond to a transition form between alpha-acrosin to beta-acrosin. In conclusion, rabbit proacrosin/acrosin system remains in the large proportion as proacrosin during capacitation and acrosome reaction.

Variaçäo da concentraçäo espermática entre amostras obtidas após curto intervalo de tempo / Variation in the eperm concentration between samples obtained with brief time interval

A probabilidade de gestaçao com o uso das técnicas de reproduçao assistida está ligada com o número de espermatozóides móveis (EM) recuperados após o preparo do esperma. Em geral, aceita-se como ideal no programa de inseminaçao intra-uterina a obtençao de no mínimo 5.0 milhoes de EM/ml, e no programa de fertilizaçao in vitro convencional 1.5 milhoes de EM/ml. A obtençao de uma segunda amostra é sempre aconselhada quando se supoe que esses valores nao sejam atingidos. Este estudo compara a concentraçao de EM a fresco, após a utilizaçao de idêntica técnica de capacitaçao (gradiente de Percoll) entre duas amostras de esperma colhidas de 20 pacientes com intervalo de uma a três horas. A concentraçao de EM/ ml no sêmen a fresco na primeira amostra foi de 2.92 + 2.86 milhoes versus 11.78 + 13.05 milhoes na segunda amostra (p = O.001, Wilcoxon). Por outro lado, a concentraçao de EM/ml após o processo de capacitaçao na primeira amostra foi de 2.81 + 2.36 milhoes versus 5.16 + 4.60 milhoes na segunda amostra (p = O.021, Wilcoxon). Em conclusao, o número de EM obtidos no sêmen a fresco e a pós-capacitaçao numa segunda amostra (p = O.021 Wilcoxon). Em conclusao, o número de EM obtidos no sêmen a fresco e a pós-capacitaçao numa segunda amostra foi significativamente superior ao encontrado na primeira amostra, justificando a conduta de se obter uma nova amostra, caso a primeira seja insuficiente para a utilizaçao das técnicas de reproduçao assistida.

Efecto protector de la albúmina sobre las membranas del espermatozoide humano en la capacitación espermática in vitro / Albumin protective effect on human spermatozoa membranes during in vitro sperm capacitation

Se estudió el efecto de la preparación in vitro de semen sobre la estructura y sobrevida del espermatozoide humano, con el objeto de valuar si la albúmina protege las membranas durante la centrifugación. Se analizaron de manera cegada y aleatoria 12 muestras seminales de hombres fértiles a las cuales se les trató por tres métodos diferentes (swim-up, gradientes de albúmina, gradientes de percoll), para separar la fracción móvil de espermatozoides. Las membranas plasmática y acrosomal externa y la sustancia acrosomal de los espermatozoides fueron observadas al microscopio electrónico de transmisión en 50 espermatozoides por muestra y la viabilidad y movilidad progresiva se analizó después de incubación durante 24 horas a 37ºC. Se presenta evidencia que sugiere que la albúmina estabiliza la membrana acrosomal externa del espermatozoide humano e incrementa la viabilidad de esta célula

Variabilidad del análisis seminal y de los parámetros objetivos del movimiento espermático en semen y durante la capacitación espermática en individuos con fertilidad probada / Variability of semen analysis and of objective parameters of sperm motion in semen and during sperm capacitation in males with probed fertility

Estudiamos la variabilidad de los parámetros seminales y del movimiento objetivo espermático en semen y a las 2 horas de capacitación a través de métodos computacionales en individuos con fertilidad probada. Analizamos la variabilidad intraindividuo en 20 hombres y la variabilidad interindividuo en 80 donantes fértiles. El 12,5 por ciento de la población estudiada presentó oligospermia pura sin otra alteración del espermiograma. Sus parámetros de movimiento celular fueron comparables a la población de donantes normospérmicos. En semen, la concentración espermática fue la que presentó mayor variación tanto intra como interdonante y la menor variación se observó para el porcentaje de motilidad y el porcentaje de formas normales. Del estudio de los parámetros objetivos del movimiento espermático destaca una menor variabilidad intra e inter donante durante la capacitación espermática comparado con el semen. Estos datos sugieren que la evaluación del movimiento espermático a las 2 horas de capacitación podría ser un método útil, adicional al espermiograma en la evaluación del potencial de fertilidad masculino

Taxas de clivagem embrionária após inseminaçäo de oócitos com espermatozóides capacitados no fluido folicular / Rate of cleavage after oocytes insemination with spermatozoa capacitated in follicular fluid

A finalidade este estudo foi comparar as taxas de clivagem de embriöes após inseminaçäo de oócitos com espermatozóides capacitados na presença ou näo de fluido folicular. No primeiro experimento, um total de 67 oócitos foram inseminados com espermatozóides que tiveram parte do procedimento de capacitaçäo com incubaçäo curta em fluido folicular (21 oócitos - experimento I) e espermatozóides capacitados com uso de apenas o meio de cultura Menezo B2 (46 oócitos - controle). Näo houve diferença (p=0.89) entre o número de embriöes obtidos: 13 (experimento I) e 29 (controle). No segundo experimento, um total de 49 oócitos foram inseminados com espermatozóides que tiveram todo o procedimento de capacitaçäo realizado com fluido folicular num tempo de incubaçäo longo (13 oócitos - experimento II) e espermatozóides capacitados com o uso de apenas o meio de cultura Menezo B2 (36 oócitos - controle). Näo houve diferença (p=0.31) entre o número de embriöes obtidos : 7 (experimento II) e 25 (controle). A ausência de diferenças entre as taxas declivagem nos dois experimentos informa que o fluido é um meio capaz de preparar adequadamente os espermatozóides, propiciando o fenômeno de capacitaçäo adequado. Contudo esta açäo do fluido näo é superior a observada com o uso de um meio de cultura comum com o Menezo B2