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Protein tyrosine phosphorylation of the human sperm head during capacitation： immunolocalization and relationship with acquisition of sperm-fertilizing ability

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Asian Journal of Andrology 2010年第6期12卷 853-861页

作者： Arcangelo Barbonetti Maria Rosaria C. Vassallo Giuliana Cordeschi Dimitrios Venetis Andrea Carboni Alessandra Sperandio Giorgio Felzani Sandro Francavilla Felice Francavilla Andrology Unit Department of Internal Medicine University of L＇Aquila Coppito 67100 L＇Aquila Italy Istituto di Ricovero e Cura a Carattere Scientifico S.Raffaele Sulmona 67039 Italy

The occurrence of tyrosine phosphorylation （TP） in the sperm head during capacitation has been poorly investigated, and no data exist on the relationship of its dynamics with the acquisition of sperm fertilizing ability. This study localized TP of head proteins in human spermatozoa during capacitation and explored its relationship with acquisition of the ability to display progesterone （P）-stimulated acrosome reactions （ARs） and to penetrate zona-free hamster oocytes. By immunofluorescence, TP immunoreactivity was revealed in the acrosomal region of formaldehyde-fixed/unpermeabilized samples, whereas it was abolished in fixed/permeabilized samples, in which TP immunoreactivity was high in the principal piece. No TP immunoreaetivity was detectable in unfixed spermatozoa. Head TP immunoreactivity was localized externally to the acrosome, close to the cytoplasmic membrane, as assessed by transmission electron microscopy. The increase in head TP was an early event during capacitation, occurring within 1 h in capacitating conditions. At this time, the P-stimulated ARs were also increased, whereas egg penetration was as poor as in uncapacitated spermatozoa. At 5 h of capacitation, the extent of neither head TP nor the P-induced ARs were greater than that at 1 h, whereas egg penetration had significantly increased. Seminal plasma inhibited head TP, P-induced ARs and egg penetration. None of these inhibitory effects, unlike those on tail TP, were prevented by the cAMP analogue dbcAMP （N,2-O-dibutyryladenosine 3＇,5＇-cyclic monophosphate）. In conclusion, head TP is a subsurface event occurring early during capacitation and is closely related to acquisition of the ability to display P-stimulated ARs, whereas the ability to fuse with oolemma and to decondense is a later capacitation-related event.

关键词： acrosome reaction capacitation human spermatozoa sperm-oocyte fusion tyrosine phosphorylation

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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 capacitation.However,structural and functional sperm changes during capacitation currently remain poorly defined.Here,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 samples.We 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 recovered.Furthermore,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 sites.Overall,the multivariate biomarkers analyzed showed a less variable distribution on spermatozoa recovered after 4 h capacitation than that with the shorter capacitation time.These 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

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Effect of low-dose fenvalerate on semen quality capacitation in adult mice

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Chinese Medical Journal 2011年第10期124卷 1529-1533页

作者： SHI Xiao-dan BI Huan-jing FU He-ling LI Liang-yun LIU De-kang LI Jian-min Laboratory of Reproductive Medicine Department of Cell Biology and Medical Genetics Nanjing Medical University Nanjing Jiangsu 210029 China

Background Fenvalerate （FEN） has been demonstrated to be a reproductive toxicant in humans and rodents. However,little is known about whether short-term exposure to low-dose FEN produces reproductive toxicity.Methods We administered FEN （0.009 375, 0.1875, 3.750, or 45.00 mg·kg-1d-1 by gavage for 30 days） to male ICR mice and compared reproductive toxicity parameters between groups receiving different concentrations of FEN.Reproductive toxicity was evaluated by computer-assisted semen quality analysis （CASA）, chlortetracycline （CTC） assay,and histopathology.Results The sperm morphology and testis histology of FEN-exposed mice （all doses） were similar to that in controlling mice. Exposure to FEN at a concentration of 0.1875 mg·kg-1d-1 decreased sperm path straightness （STR） and linearity （LIN）（both P〈 0.05）, but had no significant impact on average path velocity （VAP）, straight line velocity （VSL）, curvilinear velocity （VCL）, lateral amplitude （ALH）, beat cross frequency （BCF）, or progressive motility （MOT）. FEN reduced the rate of mouse sperm capacitation in a dose-dependent manner.Conclusion The present results demonstrate that exposure to low-dose FEN for 30 days reduces semen quality and sperm capacitation in adult mice.

关键词： fenvalerate capacitation computer-assisted sperm analysis, sperm

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STUDY ON in vitro SPERM capacitation AND EGG-PENETRATION OF GIANT PANDA

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Science China Chemistry 1989年第4期32卷 435-441页

作者：陈大元宋祥芬赵学坤段崇文叶志勇何光昕张安居冯文和 Laboratory of Reproductive Biology Institute of Zoology Academia Stnica Beijing Chengdu Research Base of Giant Panda Breeding Department of Biology Sichuan University Chengdu

A preliminary report on in vitro sperm capacitation and egg-penetration of giant panda is briefly presented. The panda spermatozoon consists of head, neck and tail, just like the spermatozoa of other animals. Before capacitation sperm heads clusterd together and dispersed after capacitation. They were then able to swim straight forwared. During the time of in vitro capacitation the plasm membrane of the sperm head was first expanded tov arious degrees, then disintegrated, and finally became detached. The electro-dense material in the acrosome appeared in small clumps with high density. Extensive vesiculation occurred between the bi-layered acrosome membranes and thus led to disintegration. Vesiculation in panda sperm differs from that reported in hamsters. When the capacitated panda spermatozoa came into contact with the hamster eggs, the region between the acrosome collar and postacrosome cap first fused with the egg membrane followed by the penetration of the nucleus into the cortex of the egg. Some

关键词： spermatozoa capacitation penetration in vitro fertilization.

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Effects of ubiquitin-protea-some pathway on mouse sperm capacitation, acrosome reaction and in vitro fertilization

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Chinese Science Bulletin 2002年第2期47卷 127-132页

作者： WANG Hongmei SONG Changcheng DUAN Chongwen SHI Weixian LI Cunxi CHEN Dayuan WANG Yongchao State Key Laboratory of Reproductive Biology Institute of ZoologyChinese Academy of SciencesBeijing 100080China Institute of Cell Biology College of Life SciencesBeijing Normal UniversityBeijing 100875China

Chlortetracycline (CTC) fluorescence patterns were used to study changes in the patterns B and AR of mouse sperm after incubation with reagents that would block the UPP. They were the monoclonal antibody againstubiquitinated proteins——UCPi; the polyclonal antibodyagainst ubiquitin-anti-Ub, and a special inhibitor againstproteasome——ALLN. Furthermore, we treated the capaci-tated sperm or the eggs with these reagents separately and tested whether the normal in vitro fertilization was blocked or not. Results illustrate that UCP1, anti-Ub, and ALLN have little effects on sperm capacitation and acrosome reaction, but they do inhibit fusion of mouse sperm with eggs, which suggests that UPP play an important role in mouse in vitro fertilization.

关键词： ubiquitin-proteasome pathway capacitation acrosome reaction in vitro fertilization.

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Role of ions and ion channels in capacitation and acrosome reaction of spermatozoa

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Asian Journal of Andrology 1999年第3期1卷 95-107页

作者： Sharad B Purohit Malini Laloraya G.Pradeep Kumar School of Life Sciences Devi Ahilya University Vigyan Bhavan Khandwa Road Indore 452001 M. P. India.School of Life Sciences Devi Ahilya University Vigyan Bhavan Khandwa Road Indore 452001 M. P. India.School of Life Sciences Devi Ahilya University Vigyan Bhavan Khandwa Road Indore 452001 M. P. India.

capacitation and acrosome reaction are important prerequisites of the fertilization process. capacitation is a highlycomplex phenomenon occurring in the female genital tract, rendering the spermatozoa capable of binding and fusionwith the oocyte. During capacitation various biochemical and biophysical changes occur in the spermatozoa and thespermatozoal membranes. Ions and ion channels also play important roles in governing the process of capacitation bychanging the fluxes of different ions which in turn controls various characteristics of capacitated spermatozoa. Alongwith the mobilization of ions the generation of free radicals and efflux of cholesterol also plays an important role in thecapacitation state of the spermatozoa. The generation of free radical and efflux of cholesterol change the mechano-dynamic properties of the membrane by oxidation of the polyunsaturated lipids and by generating the cholesterol freepatches. The process of capacitation renders the spermatozoa responsive to the inducers of the acrosome reaction. Theglycoprotein zona pellucida 3 (ZP3) of the egg coat zona pellucida is the potent physiological stimulator of the acro-some reaction; progesterone, a major compoent of the follicular fluid, is also an induce of the acrosome reaction.The inducers of the acrosome reaction cause the activation of the various ion-channels leading to high influxes of calci-um, sodium and bicarbonate. The efflux of cholesterol during the process of capacitation alters the permeablity of themembrane to the ions and generate areas which are prone to fusion and vesculation process during the acrosome reac-tion. Ths review focuses mainly on effects of the ion and ion-channels, free radicals, and membrane fluidity changesduring the process of capacitation and acrosome reaction. (Asian J Androl 1999 Sep; 1: 95-107)

关键词： spermatozoa capacitation acrosome reaction ion channels free radicals membrane fluidity

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Protein kinase inhibitors affect spermatogenic functions and blood testis barrier remodelling:A scoping review

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Asian pacific Journal of Reproduction 2023年第3期12卷 97-108页

作者： Oyovwi Mega Obukohwo Onome Bright Oghenetega Falajiki Yewande Faith Emojevwe Victor Rotu Arientare Rume Joseph Gregory Uchechukwu Oyeleke Abiodun Abioye Department of Physiology Faculty of Basic Medical SciencesAdeleke UniversityEdeOsun StateNigeria Department of Physiology School of Basic Medical ScienceBabcock UniversityIllisan-RomoOgun StateNigeria Department of Physiology University of Medical SciencesOndo StateNigeria Department of Physiology University of IbadanIbadanOyo StateNigeria Department of Medical Laboratory Science Adeleke UniversityEdeOsun StateNigeria Department of Physiology Federal University Oye-EkitiNigeria

Objective:To identify the role of protein kinase in male reproduction in animal models and human spermatogenic function.Methods:This study assessed the protein kinase of male reproduction in animal models and human using different reviewed paper indexed in PubMed,Science Direct,EBSCO,Scopus,Cochrane Library,Sage Journals,and Google Scholar.Data were charted based on author,year of publication published between 1893 and 2023,country,purpose,data collection,key findings,and research focus/domain.Results:The MAPK pathway contributed to the growth,maturation,and functionality of male germ cells.We also found out that certain influencing factors categorized into hormonal/non hormonal factors and chemotoxicant,as well as heat stress expressed an inhibitory mechanism on protein kinase,thus affecting spermatogenic functions and maintenance/remodeling of the blood testis barrier,as well as the physiology of the Sertoli cells necessary for nutritional support of spermatogenesis.However,activating protein kinases pathway like the mTOR pathway as well as increased expression of peroxiredoxin-4 and L-carnitine mediated protein kinases may be useful for treating or managing male reproductive dysfunction.Conclusions:Protein kinase plays an important role in spermatogenic functions and blood testis remodeling in animal and human.Its assessment provides essential information that can guide treatment strategies aimed at improving male reproductive potential.Taken together,these recent advances highlight a future therapeutic intervention in assessing male reproductive potential.It might also be possible to look at potential targets for male contraceptives in the MAPK pathway.

关键词： Spermatogenesis Protein kinase Inhibitors Sertoli cells capacitation mTOR

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Role of the epididymis in sperm competition

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Asian Journal of Andrology 2007年第4期9卷 493-499页

作者： Russell C. Jones Jean-Louis Dacheux Brett Nixon Heath W. Ecroyd Discipline of Biological Sciences University of Newcastle Callaghan 2308 Australia UMR INRA-CNRS 6073 Physiologie de la Reproduction et des Comportements Nouzilly 37380 France Department of Chemistry University of Adelaide Adelaide 5005 Australia

Although it is generally understood that the testes recruited kidney ducts for reproductive function during the evolution of vertebrates, little is understood of the biological significance of the adaptation. In the context of the evolution of the mammalian epididymis, this report provides evidence that a major role of the epididymis is to enhance a male＇s chance of achieving paternity in a competitive mating system. A unique example of sperm cooperation in monotremes is used as evidence that the epididymis produces sperm competition proteins to form groups of 100 sperm into bundles that have a forward motility nearly thrice that of individual spermatozoa. As it required 3-h incubation in vitro under capacitation conditions to release motile sperm from the bundles, it is suggested that the monotremes provide an example of capacitation that is quite different from capacitation in higher mammals. It is suggested that variation between species in the intensity of sperm competition could explain the variation that occurs between species in the amount of post-testicular sperm maturation and storage in the epididymis, an explanation of why the human epididymis does not play as important a role in reproduction as the epididymis of most mammals. （Asian J Androl 2007 July; 9： 493-499）

关键词： epididymis sperm maturation sperm storage capacitation sperm competition sperm cooperation

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Cellular mechanisms regulating sperm-zona pellucida interaction

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Asian Journal of Andrology 2011年第1期13卷 88-96页

作者： Andrew T Reid Kate Redgrove R John Aitken Brett Nixon Reproductive Science Group Discipline of Biological Sciences School of Environmental and Life Sciences University of Newcastle Callaghan NSW 2308 Australia ARC Centre of Excellence in Biotechnology and Development Discipline of Biological Sciences School of Environmental and Life Sciences University of Newcastle CaUaghan NSW 2308 Australia

For mammalian spermatozoa to exhibit the ability to bind the zona pellucida （ZP） they must undergo three distinct phases of maturation, namely, spermatogenesis （testis）, epididymal maturation （epididymis） and capacitation （female reproductive tract）. An impressive array of spermatozoa surface remodeling events accompany these phases of maturation and appear critical for recognition and adhesion of the outer vestments of the oocyte, a structure known as the ZP. It is becoming increasingly apparent that species-specific zona adhesion is not mediated by a single receptor. Instead, compelling evidence now points toward models implicating a multiplicity of receptor-ligand interactions. This notion is in keeping with emerging research that has shown that there is a dynamic aggregation of proteins believed to be important in sperm-ZP recognition to the regions of sperm that mediate this binding event. Such remodeling may in turn facilitate the assembly of a multimeric zona recognition complex （MZRC）. Though formation of MZRCs raises questions regarding the nature of the block to polyspermy, formation and assembly of such a structure would no doubt explain the strenuous maturation process that sperm endure on their sojourn to functional maturity.

关键词： capacitation fertilization spermatozoa sperm-zona pellucida interaction

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Effect of recombinant and native buffalo OVGP1 on sperm functions and in vitro embryo development: a comparative study

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Journal of Animal Science and Biotechnology 2018年第1期9卷 99-110页

作者： Suman Choudhary A.Kumaresan Manish Kumar Shivani Chhillar Hrudananda Malik Sudarshan Kumar Jai K.Kaushik Tirtha K.Datta Ashok K.Mohanty Animal Biotechnology Centre National Dairy Research Institute Livestock and Production Management Section National Dairy Research Institute

Background： An oviduct-specific glycoprotein, OVGP1, is synthesized and secreted by non-ciliated epithelial cells of the mammalian oviduct which provides an essential milieu for reproductive functions. The present study reports the effects of recombinant buffalo OVGP1 that lacks post-translational modifications, and native Buffalo OVGP1 isolated from oviductal tissue, on frozen-thawed sperm functions and in vitro embryo development.Results： The proportion of viable sperms was greater（P 〈 0.05） in the recombinant OVGP1-treated group compared to the native OVGP1-treated group at 2 h, 3 h, and 4 h of incubation. The proportion of motile sperms at3 h and 4 h of incubation; and membrane-intact sperms at 4 h was greater（P 〈 0.05） in the native OVGP1-treated group compared to the control and recombinant OVGP1-treated groups. The proportion of capacitated and acrosome-reacted sperms was greater（P 〈 0.05） in the native OVGP1-treated group compared to the recombinant OVGP1 group at 4 h. The rates of cleavage of embryos and their development to the blastocyst stage were greater（P 〈 0.05） in the presence of either native or recombinant OVGP1 in comparison to control at 10 μg/mL concentration as compared to 5 or 20 μg/mL.Conclusions： The study suggests that both native and recombinant OVGP1 impart a positive effect on various sperm features and in vitro embryo development. However, native OVGP1 was found to have a more pronounced effect in comparison to recombinant non-glycosylated OVGP1 on various sperm functions except viability. Hence,our current findings infer that glycosylation of OVGP1 might be essential in sustaining the sperm functions but not the in vitro embryo development.

关键词： Blastocyst capacitation Fertilization Glycoprotein In vitro fertilization Oestrus Semen Sperm

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