Associação entre proteínas do plasma seminal, motilidade e viabilidade espermática em coelhos submetidos a doping genético / Association among seminal plasma proteins, sperm motility and sperm viability in rabbits submitted to gene doping

Neste trabalho foi estudada a correlação entre o perfil proteico do plasma seminal e a motilidade e viabilidade espermática em coelhos submetidos ao tratamento com vetores de expressão contendo o gene da eritropoetina (EPO) e com EPO recombinante humana. Foram identificadas, em coelhos submetidos ao tratamento com vetor de DNA contendo o gene da EPO, duas bandas proteicas associadas a alterações na motilidade espermática - 48kDa à baixa motilidade (P<0,05) e 18kDa à alta motilidade (P<0,05) - e esse fator foi associado a maior viabilidade espermática (P<0,05). Em coelhos submetidos ao tratamento com EPO recombinante, um fator proteico, 63kDa, associou-se à alta motilidade espermática (P<0,05), enquanto dois, 26 e 40kDa, foram associados à alta viabilidade espermática (P<0,05). Esses resultados sugerem que o doping genético pode ocasionar mudanças no perfil proteico do plasma seminal, provocando alterações na motilidade e viabilidade espermática.

In this study the correlation between seminal plasma protein profile and the sperm motility and sperm viability in rabbits submitted to treatment with an expression vector containing EPO gene and with human recombinant EPO was evaluated. In rabbits submitted to treatment with EPO expression vector, two protein bands were associated to sperm motility - 48kDa associated to low motility (P<0.05) and 18kDa to high motility (P<0.05) - and this protein band was also associated to high sperm viability (P<0.05). In rabbits submitted to treatment with human recombinant EPO, a protein factor, 63kDa, was associated to high sperm motility (P<0.05) while two protein factors, 26 and 40kDa, were associated to high sperm viability (P<0.05). These results suggest that gene doping leads to changes in rabbit seminal plasma protein, altering sperm motility and sperm viability.

Alergia al plasma seminal humano: ¿mito o realidad? / Human seminal plasma allergy

Antecedentes: la hipersensibilidad al plasma seminal humano abarca una amplia variedad de manifestaciones clínicas que comprenden desde prurito local y reacciones dérmicas localizadas, hasta situaciones que ponen en riesgo la vida, como la anafilaxia. Objetivo: caracterizar este fenómeno, para el estudio a profundidad del tema y enfatizar en un problema que no está siendo valorado debido al poco conocimiento del evento. Método: revisión de la literatura empleando los términos "semen allergy" y "human seminal plasma allergy" y sus equivalentes en español en diferentes bases de datos. Resultados: este desorden inmunológico es más frecuente entre los 23 y los 35 años de edad, en la mayoría de los casos los síntomas se inician dentro de la primera hora después de culminada la relación sexual o inmediatamente después de tener contacto con el semen. El método de prevención más eficaz es el condón, aunque no es una opción adecuada para las parejas que desean concebir. Conclusión: se requiere estudiar y caracterizar mejor este fenómeno para mejorar tanto su diagnóstico como su tratamiento.

Background: human seminal plasma hypersensitivity includes a wide variety of clinical manifestations comprising itching and localized dermal reactions to situations that threaten life as anaphylaxis. Aims: to characterize this phenomenon, for in-depth study of the subject and emphasize a problem that is not being assessed due to poor knowledge of the event. Method: review of the literature using the terms "semen allergy" and "human seminal plasma allergy" and their spanish equivalents in different databases. Results: this immune disorder is more common between 23 and 35 years of age, in most cases the symptoms begin within the first hour after culminating intercourse or immediately after contact with the semen and most effective prevention method is the condom, although not an adequate solution for couples who want to conceive. Conclusion: further studies are required to further characterize this phenomenon to improve both diagnosis as treatment.

Mycoplasmas y anticuerpos anti-Chlamydia en semen de hombres infértiles y su relación con la calidad seminal y los marcadores de glándulas sexuales accesorias masculinas / Mycoplasmas and antibodies anti-Chlamydia in semen of infertile men and their relationship with seminal quality and markers of male accessory sex glands

La infertilidad masculina puede deberse a inflamación o infección del tracto genital entre otras causas. En el problema de la infertilidad masculina pueden estar implicadas las glándulas sexuales accesorias y la función espermática. En este trabajo se trata de asociar los gérmenes más frecuentes en semen de hombres infértiles incluyendo Chlamydia trachomatis, Mycoplasma hominis y Ureaplasma urealyticum con las características seminales, y los niveles de fructosa, ácido cítrico y a-glucosidasa neutra como marcadores de las glándulas sexuales accesorias masculinas. La detección de los anticuerpos indicó que C. trachomatis fue el germen de mayor prevalencia. Los anticuerpos (Acs) anti-Chlamydia, Mycoplasma hominis y Ureaplasma urealyticum se asociaron con descenso de los marcadores glandulares fructosa y a-glucosidasa. Por otra parte se observó aumento del pH y leucocitospermia en los pacientes con Acs anti-Chlamydia. La evaluación microbiológica y la bioquímica del semen orientarían más sobre la propagación de la infección y permitiría seleccionar la terapia más efectiva. Se observó que es importante la evaluación microbiológica y de los marcadores de glándulas accesorias sexuales masculinas en el semen para diagnosticar y tratar las infecciones masculinas.

Male infertility may be due to inflammation or infection of the genital tract among other causes. Male accessory sex glands and sperm function may also be involved in the problem of infertility. This study tries to associate the most frequent bacteria in semen of infertile men including Chlamydia trachomatis, Mycoplasma hominis and Ureaplasma urealyticum with the seminal characteristics and levels of fructose, citric acid and a-neutral glucosidase as markers of the accessory glands. Detection of antibodies anti Chlamydia trachomatis indicated that it was the most prevalent germ. Antibodies (Ab) anti-Chlamydia, Mycoplasma hominis and Ureaplasma urealyticum were associated with a decrease of the glandular markers fructose and a-neutral glucosidase. On the other hand, there were increased pH and leukocytospermia in men positive for antibodies anti-Chlamydia. Microbiological and biochemical evaluation of semen could orient more about the spread of infection and allow for the selection of the most effective therapy. We find that microbiological and glandular accessory markers assessments in semen are important to diagnose and to treat infections.

PCR Multiplex fluorescente para detecção de bactérias em sêmen bovino / Fluorescent Multiplex PCR for detection of bacteria in semen bovine

Este estudo teve como objetivo avaliar o limiar de detecção da técnica de PCR multiplex fluorescente aliada a eletroforese capilar na detecção de agentes infecciosos em amostras de sêmen experimentalmente contaminadas com concentrações decrescentes das bactérias Brucella abortus, Leptospira interrogans sorovar pomona, Campylobacter fetus e Haemophilus somnus. Amostras de sêmen bovino foram experimentalmente contaminadas com concentrações decrescentes de bactérias obtidas através de diluições seriadas na base 10 de modo a obter-se amostras contendo desde 1 vez até 10-7 bactérias/mL a partir da concentração inicial de Leptospira pomona, Brucella abortus, Campylobacter fetus e Haemophilus somnus. As diluições foram efetuadas individualmente para cada bactéria, bem como nas diferentes concentrações necessárias para a padronização do teste de multiplex PCR. As extrações de DNA de todas as soluções contendo espermatozóides e bactérias analisadas no presente estudo foram realizadas segundo protocolo descrito por Heinemann et al. (2000). Os produtos de PCR multiplex foram avaliados por eletroforese em gel de poliacrilamida 8% e separação eletroforética por sistema capilar em equipamento automático de análise de fragmentos de DNA MegaBace. Observou-se a amplificação de fragmentos de 193pb, 330pb, 400pb e 415pb a partir do DNA de B. abortus, L. pomona, H. somnus, C. fetus, respectivamente. Na análise por eletroforese capilar de produtos da PCR multiplex do DNA para detecção simultânea dos quatro patógenos observou-se a sinal de positividade até a diluição de 10-3 bactérias/mL vezes da concentração inicial da solução estoque de cada bactéria. A técnica de PCR multiplex aliada à eletroforese capilar foi usada pela primeira vez para o diagnóstico direto de quatro bactérias patogênicas no sêmen, demonstrando ser um método rápido na detecção de bactérias causadoras de doenças reprodutivas.

This study aimed to evaluate the threshold of detection of fluorescent multiplex PCR coupled with capillary electrophoresis for detection of infectious agents in semen samples from experimentally infected with decreasing concentrations of the bacteria Brucella abortus, Leptospira interrogans serovar pomona, Campylobacter fetus and Haemophilus somnus. Samples of bovine semen were experimentally infected with decreasing concentrations of bacteria obtained from serial dilutions in the base 10 so as to obtain samples containing a long time until 10-7 bacteria/mL from the initial concentration of Lepstospira pomona, Brucella abortus, Haemophilus somnus and Campylobacter fetus. The dilutions were made individually for each bacterium, as well as in different concentrations needed to standardize the multiplex PCR test. DNA extractions of all solutions containing sperm and bacteria analyzed in this study were performed according to protocol described by Heinemann et al. (2000). The multiplex PCR products were analyzed by electrophoresis on 8% polyacrylamide gel and capillary electrophoretic separation system for automated equipment in the analysis of DNA fragments MegaBACE. We observed amplification of fragments of 193pb, 330pb, 415pb and 400bp from the DNA of B. abortus, L. pomona, H. somnus, C. fetus respectively. The analysis by capillary electrophoresis of multiplex PCR products of DNA for simultaneous detection of the four pathogens was observed by detecting the dilution of 10-3 bacteria / mL times the initial concentration of the stock solution of each bacterium. The multiplex PCR coupled with capillary electrophoresis was first used for the direct diagnosis of four pathogenic bacteria in semen, proving to be a rapid method to detect bacteria that cause reproductive disorders.

Evaluación del Sistema Cobas Ampliprep/Cobas Amplicor HIV-1 Monitor tm test 1.5 en plasma seminal / Evaluation of the Cobas Ampliprep/Cobas Amplicor HIV-1 Monitor tm test 1.5 system in sperm samples

La determinación de la carga viral de HIV y la detección del ADN proviral en semen se utilizan en protocolos de fertilización para parejas discordantes (hombre HIV positivo - mujer AMPLICOR HIV-1 MONITOR tm Test versión 1.5 (Roche) en plasma seminal para estimar la carga viral. Se procesaron 31 muestras de plasma seminal, diluidas y sin diluir para evaluar la amplificación del control interno y descartar posibles inhibiciones. Luego se contaminaron 22 muestras con plasma de pacientes HIV positivos para verificar la cuantificación del ARN viral. Finalmente, se procesaron 12 muestras en paralelo por Nuclisens HIV-1 QT (Biomerieux) para descartar potenciales falsos negativos. Concluimos que el sistema evaluado es un método adecuado para cuantificar ARN viral en plasma seminal. No se observó inhibición, ni falsos negativos y los valores de carga viral y el límite de detección no se vieron modificados por la matriz diferente.

Analyses of the HIV load and presence of proviral DNA in sperm samples are used in assisted fertilization protocols for discordant couples (infected man-healthy woman). We evaluated the use of the COBAS Ampliprep/COBAS AMPLICOR HIV-1 MONITOR tm Test versión 1.5 (Roche) for viral load quantification in seminal plasma samples. We first tested 31 sperm samples for amplification of the internal control to discard potential inhibition. Seminal plasmas were analyzed directly and diluted. We then spiked 21 sperm samples with human plasma from HIV-positive patients to confirm that HIV RNA could be amplified. We also compared the results of 12 samples with NASBA (Nuclisens HIV-1 QT, Biomerieux), and confirmed lack of false negative results. We conclude that the new assay is an adequate methodology to analyze HIV load in sperm samples. We did not observed inhibition, neither false negative results and quantification demonstrated equivalent HIV loads.

Influence of antisperm antibodies in the semen on intracytoplasmic sperm injection outcome

OBJECTIVE: The aim of this study was to analyze the influence of autoantibodies against spermatozoa present in the semen on the outcome of in vitro fertilization with intracytoplasmic sperm injection (ICSI). MATERIALS AND METHODS: We performed a retrospective analysis of clinical and laboratorial data from a six year-period ICSI cycles. Screening for the presence of ASA in the semen, by using the direct immunobeads test (IBT), was available for 351 cycles. According to the percentage of antibody-bound spermatozoa in the semen, we divided the cycles in four groups: I (n = 194): 0 percent-10 percent ASA; II (n = 107): 11 percent-20 percent; III (n = 33): 21 percent-50 percent and IV (n = 17): 51 percent-100 percent ASA. Additionally, a group of 349 ICSI cycles performed with ejaculated spermatozoa from oligo/asthenozoospermic men who had insufficient number of motile sperm available for ASA screening was included for comparison. ICSI outcomes were compared among groups and included fertilization rate (2 PN), cleavage rate, cleavage velocity, embryo quality, clinical pregnancy and miscarriage rates. Data were examined statistically, with an alpha level of 5 percent considered significant. RESULTS: Fertilization, cleavage rate and velocity, percentage of good quality embryos, as well as clinical pregnancy and miscarriage rates did not differ among different ASA levels groups. ICSI outcomes in men exhibiting different levels of autoimmunity against spermatozoa did not differ from those with severely abnormal seminal parameters. CONCLUSIONS: Our data indicate that intracytoplasmic sperm injection (ICSI) outcomes are not influenced by ASA levels on sperm.

Role and impact of changes of seminal plasma cytokines in male infertility

The present study was aimed to measure the levels of seminal plasma cytokines including tumor necrosis factor - alpha [TNF-alpha], interferon- gamma [INF- gamma], interleukin 8 [IL-8] and interleukin 6 [IL-6] in infertile and fertile males. It is aimed also to determine the relationship of seminal cytokines with semen quality. The study was performed upon 10 normal healthy and 30 adult men complaining of primary infertility in spite of regular vaginal coitus with intra-vaginal ejaculation and the duration of marriage was more than one year ago. Cases were classified equally into 4 groups according to history taking, clinical findings and semen examination. Group 1 [control] comprised of fertile adult men with sperm count >20 x 10 6 sperm/cc. Group II was infertile adult men with varicocele and oligospermia. Group III included infertile adult men with genitourinary tract infection with oligospermia. Group IV was infertile adult men with azospermia. TNF-alpha, INF-gamma, IL-8 and IL-6 concentrations in seminal plasma were determined using high sensitivity ELISA kits specific for each cytokine separately. Seminal levels of TNF-alpha, INF-gamma, IL-8 and IL-6 were significantly higher in oligospermia with genitourinary infection than the other groups while the azospermic group showed the lowest seminal levels of cytokines. There are a significant and positive correlation between both TNF-alpha, IFN-gamma, IL-8 and some abnormal parameters in semen analysis which actually leads to infertility. These parameters are percentage of immotile sperms, sluggish sperms and abnormal forms of sperms. IL-6 was significantly correlated with leukocytic count/HPF. Seminal plasma cytokines were significantly increased in oligospermic patients with genitourinary infection. They play an important role in male infertility as there are positive correlation between their levels in seminal plasma and different abnormalities in semen analysis. Their measurement in seminal plasma seems to be a sensitive and useful marker for infection/inflammation of the genital tract

IgA class entity determination study in secretion from the mucosal immune system compartments

Five separated mucosal viscous colloidal solutions were obtained from human saliva, semen, feces, colostroum and urine. These were biurate positive, 2ME resistant with antibody activity during infection. Solutions were designated as mucosal immunoglobulins [MIG]. MIG separation were performed by using 6% polyethylene glycol solution. Rabbit standard polyclonal anti SIgA was obtained through hyperimmunization of rabbit with standard SIgA. MIG solutions were of SIgA class entity as indicated by immunoelectrophoresis and radial immunoprecipition in gel. The use of PEG 6% for separation and class entity determination of MIG are being reported for the first time in this area

Anticuerpos de la clase IgG unidos a la superficie espermática: correlación metodológica, cuadro citológico / Antisperm antibody binding on the sperm surface: statistical correlation between method, cytologycal characteristics

Entre el 5 y 10 por ciento de los hombres infértiles presentan en sus eyaculados anticuerpos antiespermatozoides (AcAe) en valores que superan las tasas aceptadas, por lo que la detección de este tipo de anticuerpos debe realizarse al iniciar los estudios de una pareja infértil. En este trabajo se emplearon eyaculados procedentes de hombres integrantes de parejas infértiles, con el objetivo de a) establecer la correlación entre dos métodos utilizados en la detección y cuantificación de AcAe con capacidad de unión a la membrana espermática, los Immunobeads test (IB) y el Mixed Antiglobulin Test: MAR test (Mt) y b) determinar las características citológicas de los eyaculados de hombres con alta tasa de anticuerpos con capacidad de unión a la membrana del espermatozoide. Para el estudio comparativo de ambos test se emplearon 58 muestras. El criterio de selección empleado es que uno de los dos tests superara los valores de referencia. La evaluación citológica se realizó en 250 muestras tomadas al azar y en 53 eyaculados leucospérmicos (concentración de leucocitos polimorfonucleares) (LPN) > 10 a la sexta. Método: Peroxidasa. Los IB para la detección de anticuerpos de clases IgG (IBG) e IgA (IBA) y Mt fueron desarrollados según normas de la OMS, empleándose la técnica directa en todas aquellas muestras con concentración espermática mayor a 5.10 a la sexta espermatozoides/mL y movilidad traslativa (grados [a] + [b]) superior a 40 por ciento, en los restantes casos se usó el método indirecto. En los 58 sémenes seleccionados, los anticuerpos de tipo IgG presentaron valores de (x+/-DS) 88,9+/-15,8 y de 58,8+/-31,6 para Mt e IBG respectivamente (P < 0,001). En el 86,2 por ciento de los casos (n=50) ambos se correlacionaron dando resultados francamente positivos, en el 13,8 por ciento (n=8) frente a IB negativos, se obtuvieron Mt positivos, no habiéndose obtenido valores dudosos en las muestras en estudio. Los valores obtenidos en los dos métodos directos que involucran a la IgG, se correlacionaron significativamente (r=0,58). Los resultados obtenidos con IB en los 58 pacientes seleccionados, mostraron en el 63,8 por ciento (n=37) de los sémenes anticuerpos IgG e IgA, en el 22,4 por ciento (n=13) sólo de clase IgG, no encontrándose ninguna muestra con anticuerpos de clase IgA, únicamente...

Immunological assessment of infertility by estimation of antisperm antibodies in infertile couples.

160 clinical samples were collected from 40 infertile couples with unexplained infertility. The samples collected included serum and seminal plasma of the male partners and serum and cervical mucus samples of the female partners. 25 fertile healthy couples were investigated as controls. All the samples collected were then tested for class-specific antisperm antibodies by an Enzyme linked immunosorbent assay (ELISA). Antisperm antibodies were detected in 30% of the infertile couples which included 25% female and 10% male partners. Amongst the cases positive for antisperm antibodies, antibodies were detected most frequently in female sera 58.4% followed by male sera 33% and 25% in cervical mucus. The isotyping of antisperm antibodies in various samples showed IgG to be the most frequent type specific antibody followed by IgM & IgA types of antibodies. ELISA has provided a relatively simple, reliable and highly reproducible method of detection of antisperm antibodies. Thus application of antisperm antibody testing especially in cervical mucus should become an integral part of the investigation of immunologic infertility.

O papel do exame do sêmen na infertilidade masculina III - testes para função espermática / The role of semen analysis in the male infertility III - Sperm function tests

O autor revisa o papel do exame do sêmen (espermograma) na investigação de infertilidade masculina. Nesta terceira parte aborda o uso de testes para avaliar a capacidade de fertilização dos espermatozóides (testes funcionais), dando ênfase às metodologias disponíveis para esta avaliação. Simultaneamente descreve alguns aspectos estruturais dos espermatozóides e conceitos atuais sobre fertilização humana.

Deteccçäo de anticorpos antiespermatozóides da classe IGA no sêmem fresco pela técnica do MAR teste (Mixed Antiglobulin Reaction) / Detection of antisperm antibodies of IGA classes on the fresh semen, with the mixed antiglobulin reaction (MAR test)

O objetivo deste trabalho foi introduzir no Laboratório de Reproduçäo Humana do Hospital das Clínicas, da Faculdade de Medicina da UFMG, o MAR Teste IgA; investigar a presença de anticorpos antiespermatozóides da classe IgA no sêmem; comparar duas técnicas para realizar o mesmo exame, (Sigma e Behring); estabelecer um ponto de corte relevante para positivar amostras e comparar os resultados com a análise seminal de rotina. Foram analisadas amostras de sêmem, através do MAR teste IgA em 216 homens, distribuidas em dois grupos; férteis; com filhos menores de dois anos e inférteis; homens provenientes de relaçäo conjugal infértil. No ponto de corte de 10 por cento, o MAR Teste IgA Behring, detectou amostras positivas em sete homens (16,7 por cento) do grupo fértil e em 57 (32,6 por cento) do grupo infértil, (p*=0,04) o MAR Teste IgA Sigma, detectou amostras positivas em quatro homens (9,8 por cento) do fértil e em 45 (26,0 por cento) do grupo infértil, (p*=0,03). No novo ponto de corte, o MAR Teste IgA Behring detectou 20 (11,4 por cento) no grupo infértil (p*=0,09). O MAR Teste IgA Sigma, detectou uma amostra positiva nos homens do grupo fértil (2,4 por cento) e 16 (9,3 por cento) do grupo infértil, (p*=0,21). As conclusöes apontam um melhor desempenho do MAR Teste IgA "Behring" como método de "screening" para infertilidade auto-imune, assim como a presença da relaçäo entre exames positivos com a contagem baixa de espermatozóides, a motilidade alterada e a viscosidade aumentada.

Relationship between the direct mixed antiglobulin reaction (MAR) test and spontaneous sperm agglutination in men from infertile couples.

To determine the relationship between the direct MAR test and spontaneous sperm agglutination in men from infertile couples, semen samples from 160 men were studied. Spontaneous sperm agglutination was present in 80 samples, and absent in the others. Routine semen analysis and the direct MAR test were performed on all samples. The incidence of positive MAR tests in samples with and without spontaneous sperm agglutination were 15 and 0 per cent respectively (p < 0.05). There were no significant differences in mean sperm concentration, motility, and morphology between the samples with and without spontaneous sperm agglutination, and between the positive and negative MAR test groups. It can be concluded that spontaneous sperm agglutination relates significantly to antisperm antibodies detected by the direct MAR test, and this factor could be used as indication for antisperm antibody testing of infertile men.

Solubilización de la membrana de espermatozoide humano con n-dodecil sarcosinato de sodio: II. Estudios sobre el extracto / Spermatozoa submitted to ionic detergent treatment: studen on the sodium n-dodceyl sarcosinate extract

El n-dodecil sarcosinato de sodio produce la solubilización del espermatozoide humano con algo de contaminación acrosomal. El extracto está compuesto por una mezcla de productos con pesos moleculares y características inmunológicas similares a las del plasma seminal humano. El hecho de que la antigenicidad se mantenga luego de la solubilización, hace que ésta sea un comienzo útil en el aislamiento de moléculas que juegan un papel en la infertilidad por causas inmunológicas

Seminal inhibin--prospects for immunocontraception.

Passive immunization of adult rats, hamsters and marmosets with rabbit anti-seminal inhibin resulted in complete or partial block of fertility. The antiserum treatment presumably neutralized endogenous inhibin resulting in an unopposed rise in circulating FSH. This probably led to a refractoriness of the testes to FSH resulting in complete spermatogenic arrest. Nevertheless, there was no change in the mating behaviour of the animals. The antibodies also affected the epididymal spermatozoa by causing large scale agglutination.

Anticorpos antiespermatozóides: imunologia das respostas auto e iso-imune / Antispermatozoa antibodies: immunology of auto and iso-immune responses

O autor faz consideraçöes sobre fatores imunológicos envolvidos no processo de formaçäo de anticorpos antiespermatozóides no homem e na mulher. Säo revisados a origem e a inibiçäo da antigenicidade espermática, os mecanismos das respostas auto e iso-imune e as células que participam destes mecanismos. Säo também descritas a imunoglobulinas detectadas no homem e na mulher na resposta imune e seus efeitos imunológicos contra os espermatozóides

Characteristics of monoclonal antibodies reactive with human sperm and seminal plasma.

The characteristics of monoclonal antibodies developed against human spermatozoa are described. Out of 10 monoclonal antibodies 9 did not react in ELISA with human RBC, WBC, platelets, Raji cells nor mouse sperm. Four monoclonal antibodies reacted with monkey sperm and all 10 reacted with human seminal plasma. Monoclonal antibodies showed differential reactivity with pre- and post-capacitated sperm. Four monoclonal antibodies were able to agglutinate sperm whereas none of these were positive in sperm-immobilization assay. Interestingly, two monoclonal antibodies (MA-46 and MA-50) were able to block the attachment of pre-capacitated sperm to zona denuded hamster oocytes. MA-46 and MA-50 recognized in immunoblot spermatozoa antigens having apparent molecular weights of 14 and 20 K Da and greater than 200 K Da respectively. The monoclonal antibodies reported in this study will be useful in further delineating the spermatozoa antigens involved in regulation of fertility.