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Changes on membrane integrity and ultrastructure of human sperm after freeze-drying

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Journal of Reproduction and Contraception 2016年第2期27卷 76-81页

作者： Wei-jie ZHU Jing LI Luan-juan XIAO Department of Developmental and Regenerative Biology College of Life Science and Technology Jinan University Department of Pathophysiology Medical College Jinan University

Objective To observe changes on membrane integrity and ultrastructure of human sperm after *** Semen samples from both normospermic donors（group A, n=15） and infertile men with abnormal sperm parameters（group B, n=15） were enrolled into this study. These samples were freeze-dried by using a freeze-drying method. The membrane integrity in the head and tail regions of individual spermatozoon was examined by using the combined hypo-osmotic swelling-eosin Y exclusion test. Sperm ultrastructure in groups A（n=3） and B（n=3） was observed by scanning electron microscopy（SEM）.Results After freeze-drying, all spermatozoa were types I（damaged both head and tail membranes） and III（damaged head membrane and intact tail membrane） membrane integrity in groups A and B. Type III of group B had lower value than that of group A（P〈0.01）. Under SEM, intact freeze-dried spermatozoa including abnormal morphology and normal-looking morphology were observed in both groups A and B. A few freezedried sperm heads had unsmooth or fuzzy surface. Isolated sperm heads, bent tails,broken sperm tails or fragmentary tails were more frequently seen in group B than those in group *** Freeze-dried human spermatozoa could have intact structural components. However, freeze-drying resulted in severe damage on membrane integrity and ultrastructure of sperm. Samples from infertile men would have less resistance to freeze-drying.

关键词： human sperm ultrastructure membrane integrity freeze-drying lyophilization

Evaluation on the Morphology and membrane integrity of Immotile Human Sperm

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Journal of Reproduction and Contraception 2006年第2期17卷 80-84页

作者： Wei-Jie ZHU Jing LI Center for Reproductive Immunology Research Jinan University Guangzhou 510632china Dept. of Patkopkysiology Medical College Jinan University Guangzhou 510632 China

Objective To determine the membrane integrity in the head and tail regions of individual spermatozoon, and observe sperm morphology for samples with totally immotile sperm. Methods Ten infertile men with immotile sperm were enrolled into this study （group A）. The membrane integrity in the head and tail regions of individual spermatozoon of immotile sperm was examined by using the combined hypo-osmotic swelling-eosin Y exclusion test （HOS-EY test）. Sperm morphology was observed by light, scanning and transmission electron microscopy. Ten semen samples from normospermic donors were used as the control （group B）. Results The percentage of sperm with intact both head and tail membranes in group A was significantly lower than that in group B （P〈0.01）, whereas the value of sperm with defective head membrane but intact tail membrane in group A was significantly higher than that in group B （P〈0.01）. Abnormal sperm morphology in group A had a high incidence, and immotile sperm with viability and normal morphology could be observed in some cases. Most sperm had multiple ultrastructural defects. Conclussion Some immotile sperm had intact tail membrane but defective head membrane. Immotile sperm with viability and normal morphology could exist in some cases though abnormal sperm were in a great proportion. Carefully evaluating immotile sperm membrane integrity and morphology should benefit the treatment of patients with immotile sperm.

关键词： human sperm immotility membrane integrity morphology

Influence of in vitro capacitation time on structural and functional human sperm parameters

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Asian Journal of Andrology 2020年第5期22卷 447-453页

作者： Paula Saez-Espinosa Natalia Huerta-Retamal Laura Robles-Gomez Manuel Aviles Jon Aizpurua Irene Velasco Alejandro Romero Maria Jose Gomez-Torres Department of Biotechnology Faculty of ScienceUniversity of AlicanteAlicante 03080Spain FISABI0-University Hospital of San Juan de Alicante Service of Gynecology and ObstetricsSan Juan de Alicante 03550Spain Departm ent of Cell Biology and Histology Faculty of MedicineUniversity of Murcia and IMIB-ArrixacaMurcia 30100Spain IVF Spain Reproductive MedicineAlicante 03540Spain University Hospital of San Juan de Alicante Service of Gynecology and ObstetricsSan Juan de Alicante 03550Spain Human Fertility Cathedra University of AlicanteAlicante 03080Spain.

A cascade of dramatic physiological events is linked to the sperm acrosome reaction and binding to the oocyte's zona pellucida during human sperm ***,structural and functional sperm changes during capacitation currently remain poorly ***,we performed a multibiomarker approach based on the utilization of sperm concentration,motility,viability,morphology,acrosome reaction,tyrosine phosphorylation,DNA fragmentation,and lectin-binding sites to analyze the impact caused by swim-up selection times(uncapacitated,1 h capacitated,and 4 h capacitated)on sperm function and structure in normozoospermic *** found that a 4 h swim-up capacitation increased sperm quality,because a large number of cells with normal morphology and lower DNA fragmentation rates were ***,the long-term capacitation induced a higher percentage of cells with tyrosine phosphorylation of the principal piece as well as a redistribution of lectin-binding ***,the multivariate biomarkers analyzed showed a less variable distribution on spermatozoa recovered after 4 h capacitation than that with the shorter capacitation *** findings stress the importance of capacitation time as a relevant factor in sperm quality with potential biological reproductive implications both for basic research and in assisted reproduction techniques.

关键词： acrosome reaction capacitation DNA fragmentation glycocalyx lectin-binding sites membrane integrity protein phosphorylation sperm selection

Single Layer Centrifugation with Androcoll-E^(TM) improved progressive motility and percentage of live spermatozoa with intact acrosome of chilled stallion semen but did not have an effect on DNA integrity

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Open Journal of Animal Sciences 2012年第3期2卷 159-165页

作者： Ana Luisa Costa Ana Martins-Bessa Ana Rebello de Andrade Tiago Guimaraes Maria Rosa Rebordao Sandra Gamboa Pedro Pinto Bravo Maria José Correia Jorge Colaco Isabel Gaivao António Rocha Centro de Ciencia Animal e Veterinária(CECAV) University of Trás-os-Montes e Alto DouroQuinta de PradosVila RealPortugal Veterinary Degree Universty of Tras-os-Montes e Alto DouroQuinta de PradosVila RealPortugal Centro de Estudos em Ciencia Animal(CECA) Instituto de Ciencias Biomedicas Abel Salazar(ICBAS)University of PortoOportoPortugal Coimbra Higher School of Agriculture Sao Martinho do BispoCoimbraPortugal Alter Real Foundation Alter do ChaoPortugal

A significant number of stallions produce low quality ejaculates with high sensibility to chilling. Single Layer Centrifugation (SLC) with Andro-coll-ETM has been presented as an efficient method of selecting good quality spermatozoa. In the current study, changes in sperm quality (motility, viability, acrosome integrity and DNA damage) occurring during storage at 5?C for a maximum of 72 h, were investigated. For that, one ejaculate from 12 stallions was split in two aliquots: control and SLC-selected. Both aliquots were chilled and stored at 5?C and spermatozoa were evaluated for motility, viability and acrosome integrity at 24, 48 and 72 h post collection. DNA damage was evaluated at 48 h post collection using the comet assay. In the SLC-selected aliquots, there was a significant improvement in terms of progressive motility (0 h: P = 0.005;24 h: P 0.05). SLC with Androcoll-ETM improved semen quality prolonging sperm longevity of chilled semen (P = 0.012). This positive effect was more evident in ejaculates most sensitive to chilling that had a sharp decrease in motility in the first 24 h of refrigeration and for all ejaculates at 72 h post-chilling. Therefore, this method reveals to be a useful technique for selecting spermatozoa and maintain sperm quality during storage.

关键词： Stallion Sperm Single Layer Centrifugation Androcoll-ETM membrane integrity Comet assay Component Formatting Style Styling

In-vitro effect of Peganum harmala total alkaloids on spermatozoa quality and oxidative stress of epididymal ram semen

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Asian pacific Journal of Reproduction 2021年第5期10卷 232-238页

作者： Hanane Derbak Mohamed Moussaoui Amine Benberkane Abdelhanine Ayad Department of Biological Sciences of the Environment Faculty of Nature and Life SciencesUniversitéde Bejaia06000 BejaiaAlgeria

Objective:To determine the in-vitro effect of the total alkaloid extract of Peganum(P.)harmala seeds on ram epididymal ***:Semen was divided into six groups according to the following concentrations of the *** total alkaloids:1,5,10,50,and 100μg/mL,and the control *** samples were incubated at ambient temperature(21℃-24℃)for 24 h,and analyzed in terms of motility,membrane integrity,and oxidative ***:The sperm kinematic parameters,***-line velocity,curvilinear velocity,average path velocity,were significantly higher when treated with *** at concentrations ranging from 1 to 10μg/mL compared to the control group(Pmembrane integrity of ram epididymal sperm was concentration-dependent and significantly different compared to the control group(P0.05).Conclusions:Low concentrations(1-10μg/mL)of *** extracts stimulate sperm motility,preserve membrane integrity and protect ram spermatozoa from lipid peroxidation.

关键词： Alkaloids Epididymal sperm Motility Peganum harmala Ram Semen membrane integrity Lipid peroxidation 1.Introduction

Effect of low-density lipoproteins, spermatozoa concentration and glycerol on functional and motility parameters of bull spermatozoa during storage at 4℃

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Asian Journal of Andrology 2011年第2期13卷 281-286页

作者： Oscar Vera-Munoz Lamia Amirat-Briand Djemil Bencharif Marc Anton Serge Desherces Eric Shmitt Chantal Thorin Daniel Tainturier Laboratory of Biotechnology and Pathology of Reproduction ONIRIS Nantes-Atlantic College of Veterinary Medicine and Food Science 44307 Nantes France Facultad deCiencias Veterinarias Instituto de Reproduccion Animal Abraham Hemndez Prado Universidad Centra de Venezuela Maracay 2101 Aragua Venezuela Groupe Physico-chimie des Emulsions Laboratoire d＇Etude des Interactions des Molecules Alimentaires Institut National de la Recherche Agronomique 44316 Nantes Cedex 3 France MVTechnologies I＇Aigle 61300 France

An extender has been developed with low-density lipoproteins （LDLs） that eliminates the microbial risks associated with the use of whole egg yolk. The objective of this study was to assess the effects of substituting egg yolk with LDLs for use as an extender in sperm preservation at 4 ℃, as well as on spermatozoa motility, plasma membrane and acrosome integrity, at two different concentrations （80×10^6 and 240× 10^6 sperm per ml） for 8 days and to evaluate glycerol toxicity in both extenders. A total of 12 ejaculates were collected from three bulls. Spermatozoa motility was examined using computer-assisted semen analysis. Plasma membrane integrity was determined using the hypo-osmotic swelling test and acrosome integrity with the fluorescein isothiocyanate-Pisum sativum agglutinin test. The semen was subsequently divided into four aliquots and diluted with Tris-egg yolk-glycerol （TEG）, Tris-egg yolk without glycerol （TE）, LDL with glycerol （LDL＋） and LDL without glycerol （LDL-）, at 80×10^6 and 240 ×10^6sperm per ml. This study showed that the LDL＋ and LDL- extenders were more effective at preserving spermatozoa motility, plasma membrane integrity and acrosome integrity than TEG and TE （P〈0.05） during 8 days of incubation. After 3 days of incubation, a toxicity of glycerol was observed in TEG, whereas no significant difference was observed between LDL＋ and LDL-. We can therefore conclude that the LDL extender can be used to refrigerate semen at 4 ~C instead of TEG and TE at 80×10^6and 240×10^6 sperm per ml for elite bulls. This finding can be used to define a policy for the storage of high-quality bull semen.

关键词： bull semen glycerol low-density lipoproteins membrane integrity motility refrigeration