ABSTRACT
Objetivo: Describir y comparar las alteraciones ultraestructurales que puede provocar la inmovilización espermática previa a la microinyección intracitoplasmática de espermatozoides, así como los daños en el ADN y el estado del acrosoma en espermatozoides de sujetos teratozoospérmicos y normozoospérmicos. Material y métodos: Se utilizaron 15 muestras seminales procedentes de donantes normozoospérmicos y 20 muestras de pacientes teratozoospérmicos del Instituto Bernabeu de Alicante. Para el estudio con microscopia electrónica de transmisión se utilizaron ovocitos humanos como receptáculo de los espermatozoides. La fragmentación del ADN se valoró mediante la técnica TUNEL, y el estado del acrosoma, utilizando la lectina Pisum sativum conjugada con FITC. El análisis estadístico entre los diferentes grupos se realizó mediante un test ANOVA. Resultados: Los resultados mostraron que, tras la inmovilización, los espermatozoides procedentes tanto de sujetos normozoospérmicos como de teratozoospérmicos sufrieron las mismas alteraciones ultraestructurales a nivel de la membrana plasmática y acrosomal. Tras valorar la fragmentación de ADN mediante TUNEL observamos que en los espermatozoides inmovilizados el porcentaje de espermatozoides con ADN fragmentado era similar en ambos grupos (normozoospérmicos vs. teratozoospérmicos). En cambio, el porcentaje de espermatozoides con reacción acrosómica fue significativamente más elevado en los inmovilizados que en el grupo control, tanto en los sujetos normozoospérmicos como teratozoospérmicos. Conclusión: Los resultados de este estudio evidencian que los danos estructurales provocados por la inmovilización, en espermatozoides procedentes de sujetos normozoospérmicos y teratozoospérmicos, son independientes de la morfología espermática. Y además, dicho proceso no provoca fragmentación del ADN. Sin embargo, la inmovilización tanto en el grupo de los sujetos normozoospérmicos como teratozoospérmicos provoca una inducción mecánica de la reacción acrosómica (AU)
Objective: To describe and compare the ultrastructural alterations, DNA fragmentation and acrosome status that could cause sperm immobilisation prior to intracytoplasmic sperm injection, in samples from normozoospermic and teratozoospermic males. Material and methods: For this study we used 15 sperm samples from consenting normozoospermic donors and 20 samples from teratozoospermic males, submitted for assisted reproduction at Instituto Bernabeu of Alicante. To assess the ultrastructural alterations induced by immobilisation, human oocytes were used as containers for spermatozoa and then observed by transmission electron microscopy. DNA fragmentation was assessed using TUNEL and acrosome status using fluorescein isothiocyanate-conjugated Pisum sativum agglutinin. The control group consisted of sperm without manipulation. Statistical analysis between groups was performed by ANOVA. Results: The results showed that immobilised spermatozoa presented the same damage in plasma and acrosomal membranes in the samples from both normozoospermic and teratozoospermic subjects. After assessing DNA fragmentation in the cells from normozoospermic and teratozoospermic patients by the TUNEL technique, we observed that the percentage of spermatozoa with DNA fragmentation did not increase in the immobilised group compared with the control group. However, the percentage of acrosome-reacted sperm was significantly greater in immobilised spermatozoa than in the control group in both normozoospermic and teratozoospermic males. Conclusion: The results of this study show that the structural damage caused by immobilization in spermatozoa from teratozoospermic and normozoospermic males is independent of sperm morphology. In addition, the immobilisation does not cause DNA fragmentation. However, the percentage of acrosome-reacted sperm was greater in the immobilised group than in the control group, in both normozoospermic and teratozoospermic subjects (AU)
Subject(s)
Humans , Male , Sperm Injections, Intracytoplasmic/methods , Spermatozoa/ultrastructure , Sperm Immobilizing Agents/adverse effects , Sperm Motility , Acrosome Reaction , DNA FragmentationABSTRACT
OBJECTIVE: To determine the effects of lead and zinc administration on the quality of semen of albino rats. STUDY DESIGN: Experimental study. PLACE AND DURATION OF STUDY: Basic Medical Sciences Institute, Jinnah Postgraduate Medical Centre, Karachi, from August 2003 to December 2005. METHODOLOGY: Sixty adult albino rats selected for the study were divided into three groups, group A received injection normal saline 1 ml intraperitoneally daily for 8 weeks. Group B received lead chloride in a dose of 10 mg/kg body weight intraperitoneally daily. Group C received lead chloride in a dose of 10 mg/kg body weight and zinc chloride in a dose of 1 mg/kg body weight intraperitoneally daily. On the day of completion of treatment, the animals were sacrificed; epididymis was used for semen analysis. Student's t-test was used to determinate significance; the p-value < 0.05 was taken as significant. RESULTS: The number of sperms was 7.3, 1.7 and 6.6 million cells/ml in groups A, B and C respectively. Sperm's count decreased by 87% in group B (p < 0.001, CI 4117082.4 - 6965747.6) as compared to group A. Compared with group C, the sperm's count was decreased to 75% (p < 0.001, CI -5417413 to -4416987). The immotility of sperms was 27%, 57% and 26% in groups A, B and C respectively. There was 30% decreased motility of sperm in group B (p < 0.001, CI -30.19425 to -19.80575) as compared to group A. Compared with group C, the immotile sperm were increased to 31% (p < 0.001, CI 19.87494 - 30.92506). CONCLUSION: Lead produced toxic effects on germinal epithelium and altered the quality of semen which was improved by zinc.
