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Phosphodiesterase 10A inhibitor PF-2545920 as a prospective agent for the clinical promotion of sperm motility

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Asian Journal of Andrology 2023年第5期25卷 608-615页

作者： Yi-Ting Yang Bin Yan Yu-Hua Li Li-Na Guo Wei-Wei Wang Li-Jie Liu He-Guo Yu Hua Diao NHC Key Laboratory of Reproduction Regulation(Shanghai Institute for Biomedical and Pharmaceutical Technologies) School of Basic Medical SciencesFudan UniversityShanghai 200032China Reproductive Medicine Centre Zhongshan HospitalFudan UniversityShanghai 200032China School of Basic Medical Sciences Fudan UniversityShanghai 200032China Department of Urology Huashan HospitalFudan UniversityShanghai 200040China

Phosphodiesterase(PDE)inhibitors can improve sperm motility in patients with ***,the most commonly reported nonselective PDE inhibitor pentoxifylline and PDE5 inhibitor sildenafil have the disadvantages of requiring a high concentration and destroying sperm *** examined the PDE10A inhibitor PF-2545920 to compare its ability to promote sperm motility with that of pentoxifylline and *** seminal plasma was discarded,several semen samples were subjected to four treatments(control,PF-2545920,pentoxifylline,and sildenafil)to evaluate their ability to affect motility,viability,and spontaneous acrosome *** calcium and adenosine triphosphate(ATP),mitochondrial membrane potential,and penetration through viscous medium were assessed by flow cytometry,luciferase,and hyaluronic acid after treatment with *** analyses were performed using the analysis of variance statistical ***-2545920 elevated the percentage of motile spermatozoa compared to the control,pentoxifylline,and sildenafil groups at 10μmol l^(-1)(Pspermatocytes cells and spermatozoa and causes fewer spontaneous acrosomal reactions(Psperm hyaluronic acid penetrating ability(Pspermmotility.

关键词： pentoxifylline phosphodiesterase 10A sildenafil sperm motility spontaneous acrosome reaction

Changes in human sperm motility and DNA fragmentation index after incubation at different temperatures following density gradient centrifugation and swim-up procedures

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Reproductive and Developmental Medicine 2022年第4期6卷 243-248页

作者： Yan-Nan Yang Ling Wang Yu-Bing Liu Yun-Jing Xue Chen-Chen Liu Fei Shi Xue Dai Ri-Cheng Chian Center for Reproductive Medicine Shanghai 10th People’s Hospital of Tongji UniversityShanghai 200072China

Objective:The purpose of this study was to evaluate the sperm motility and DNA integrity at different temperatures to analyze whether the sperms are suitable on the second day for insemination of in vitro matured oocytes by intra-cytoplasmic sperm injection(ICSI)following density gradient centrifugation(DGC)and swim-up(SU)***:Semen samples were collected from 30 outpatients who visited the Center for Reproductive Medicine for semen *** sperm selection by DGC and SU procedures,the liquified semen samples were divided into three groups and incubated at 4,25,and 37°C,*** incubation for 24,48,and 72 hours,the sperm motility and sperm DNA fragmentation index(DFI)were ***:Following the combination of DGC and SU procedures,the sperm motility(91.8%±8.6%vs.50.8%±13.1%)and DFI(5.1%±7.9%vs.13.0%±11.6%)were significantly improved(Psperm motility of the 3 groups significantly declined(Psperm motility significantly increased(76.9%±10.4%)(Psperm DFI significantly increased(Psperm DFI did not significantly increase when the sperm samples were incubated at 4(5.7%±5.9%)and 25°C(6.8%±5.6%)for 24 hours compared to that before incubation(5.1%±7.9%).Conclusions:These results indicate that the sperm quality,in terms of motility and DFI,can be efficiently improved by DGC in combination with *** which,the sperm samples can be incubated at 25°C and be used on the second day for insemination of in vitro matured oocytes by ICSI.

关键词： sperm motility Density gradient centrifugation Swim-up DNA fragmentation index matured oocytes

Toxic effects of polychlorinated biphenyls （Aroclor 1254） on human sperm motility

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Asian Journal of Andrology 2017年第5期19卷 561-566页

作者： Li-Gang Jiang Lai-Yang Cheng Shu-Hui Kong Yang Yang Yan-Jun Shen Chao Chen Xiao-Hui Deng Shu-Zhen Liu Lan Chao Infertility Center Qilu Hospital of Shandong University Jinan 250014 Shandong China College of Life Science Shandong University Jinan 250014 Shandong China Key Laboratory of Animal Resistance Research College of Life Science Shandong Normal University Jinan 250014 Shaedong China

Polychlorinated biphenyls （PCBs） are common environmental contaminants that represent a considerable risk to reproductive toxicity in exposed human populations. Although some experimental studies have suggested an association between the levels of PCBs and semen quality, the direct effects of PCBs on human sperm parameters remain largely unexplored. To this aim, a short-term in vitro incubation experiment that better imitated the putative exposure of sperm to Aroclor 1254 （a commercial PCB mixture） in male reproduction tissue was conducted. Human sperm were incubated with various concentrations （0, 1, 5, or 25 mg ｜^-1） of Aroclor 1254 for different amounts of time （3 and 6 h） in vitro. sperm motility parameters were analyzed with computer-assisted sperm analysis （CASA）. The proportion of sperm with high mitochondrial membrane potential （ΔΨm） and the levels of intracellular reactive oxygen species （ROS） were detected to explore the probable cause of sperm impairment. Human sperm exposed to continuous Aroclor 1254 exhibited：（i） reduced sperm motility and kinematic parameters, （ii） a proportion of sperm with high ΔΨm that decreased in a dose-dependent manner （P 〈 0.05）, and （iii） increased levels of ROS compared with controls （P 〈 0.05）. In conclusion, Aroclor 1254 can decrease sperm motility, which may culminate in increased ROS and general mitochondrial dysfunction, thus affecting the fertilization potential of sperm. Our findings suggest a broader understanding of the effect of Aroclor 1254 on human soerm.

关键词： Aroclor 1254 CASA human sperm mitochondrial membrane potential reactive oxygen species sperm motility

Major regulatory mechanisms involved in sperm motility

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Asian Journal of Andrology 2017年第1期19卷 5-14页

作者： Rute Pereira Rosalia Sa Alberto Barros Mario Sousa Department of Microscopy Laboratory of Cell Biology Institute of Biomedical Sciences Abel Salazar University of Porto (ICBAS-UP) Rua Jorge Viterbo Ferreira 2284050-313 Porto Portugal and Multidisciplinary Unit for Biomedical Research-UMIB ICBAS-UP Centre for Reproductive Genetics Alberto Barros Av. do Bessa 2401° Dto. Frente 4100-012 Porto Portugal Department of Genetics Faculty of Medicine University of Porto. Alameda Prof. Hernani Monteiro 4200-319 Porto Portugal and Institute of Health Research an Innovation (13S) University of Porto Portugal.

The genetic bases and molecular mechanisms involved in the assembly and function of the flagellum components as well as in the regulation of the flagellar movement are not fully understood, especially in humans. There are several causes for sperm immotility, of which some can be avoided and corrected, whereas other are related to genetic defects and deserve full investigation to give a diagnosis to patients. This review was performed after an extensive literature search on the online databases PubMed, ScienceDirect, and Web of Science. Here, we review the involvement of regulatory pathways responsible for sperm motility, indicating possible causes for sperm immotility. These included the calcium pathway, the cAMP-dependent protein kinase pathway, the importance of kinases and phosphatases, the function of reactive oxygen species, and how the regulation of cell volume and osmolarity are also fundamental components. We then discuss main gene defects associated with specific morphological abnormalities. Finally, we slightly discuss some preventive and treatments approaches to avoid development of conditions that are associated with unspecified sperm immotility. We believe that in the near future, with the development of more powerful techniques, the genetic causes of sperm immotility and the regulatory mechanisms of sperm motility will be better understand, thus enabling to perform a full diagnosis and uncover new therapies.

关键词： antioxidants calcium membrane channels protein kinases sperm genetic abnormalities sperm motility

Chloride channels are involved in sperm motility and are downregulated in spermatozoa from patients with asthenozoospermia

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Asian Journal of Andrology 2017年第4期19卷 418-424页

作者： Shan-Wen Liu Yuan Li Li-Li Zou Yu-Tao Guan Shuang Peng Li-Xin Zheng Shun-Mei Deng Lin-Yan Zhu Li-Wei Wang Li-Xin Chen Department of Pharmacology Medical College Jinan University Guangzhou China Department of Pathology and Pathophysiology Medical College Jinan University Guangzhou China Department of Physiology Medical College Jinan University Guangzhou China Male Reproductive Center Family Planning Special Hospital of Guangdong Guangzhou China

Human spermatozoa encounter an osmotic decrease from 330 to 290 mOsm I-z when passing through the female reproductive tract. We aimed to evaluate the role of chloride channels in volume regulation and sperm motility from patients with asthenozoospermia. spermatozoa were purified using Percoll density gradients. sperm volume was measured as the forward scatter signal using flow cytometry. sperm motility was analyzed using computer-aided sperm analysis （CASA）. When transferred from an isotonic solution （330 mOsm I-z） to a hypotonic solution （290 mOsm I-Z）, cell volume was not changed in spermatozoa from normozoospermic men; but increased in those from asthenozoospermic samples. The addition of the chloride channel blockers, 4,4＇-diisothiocyanatostilbene-2,2＇- isulfonic acid （DIDS） or 5-nitro-2-（3-phenylpropylamino） benzoic acid （NPPB） to the hypotonic solution caused the normal spermatozoa to swell but did not increase the volume of those from the asthenozoospermic semen. DIDS and NPPB decreased sperm motility in both sets of semen samples. The inhibitory effect of NPPB on normal sperm motility was much stronger than on spermatozoa from the asthenozoospermic samples. Both sperm types expressed CIC-3 chloride channels, but the expression levels in the asthenozoospermic samples were much lower, especially in the neck and mid-piece areas. spermatozoa from men with asthenozoospermia demonstrated lower volume regulating capacity, mobility, and CIC-3 expression levels （especially in the neck） than did normal spermatozoa. Thus, chloride channels play important roles in the regulation of sperm volume and motility and are downregulated in cases of asthenozoospermia.

关键词： asthenozoospermia chloride channels CIC-3 sperm motility volume regulation

Inhibition of sperm motility does not affect live-dead separation of bull sperm by glass beads

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Asian Journal of Andrology 2001年第3期3卷 193-198页

作者： Robert H.Foote Department of Animal Science Cornell University Ithaca NY 14853-4801 USA

Aim: This study was designed to explore factors which influence binding of dead versus live sperm to glass ***: Multiple semen collections from bulls were used to explore selective filtration of bull sperm as influencedby nonlethal inhibition of sperm motility with fluoride, killing of sperm by quick-freezing, alteration of the glass sur-face with silicone, and different intervals of sexual rest between semen collections. Results: A comparison of glassspheres 100, 200 and 390μm in diameter indicated that 200 μm spheres were optimal for selective filtration. Quantita-tive separation of live from dead sperm was demonstrated with a correlation between the percentage of motile sperm andretention of sperm by the filter of r = -0.87 (P sperm retained by the filter despite inhibiting sperm motility during filtration, an inhibition which was reversible. Pro-portions of live-dead sperm, based upon eosin staining, were unaffected by fluoride. Coating the glass spheres with sil-icone greatly reduced selective filtration. Dead sperm adherence to glass was reduced and resistance to NaFl inhibitionwas increased by daily ejaculation versus one-week intervals of sexual rest. Conclusion; These studies indicate thatthe adherence of sperm to glass is primarily due to some form of physico-chemical change accompanying death of thesperm cell independent of active sperm motility. This attraction between the sperm plasma membrane and glass is modi-fied by the age of the ejaculated sperm. This information is useful in evaluating different clinical procedures used forsperm separation. (Asian J Androl 2001 Sep; 3: 193-198)

关键词： sperm motility glass spere filters silicone fluorides inhibition

Protein phosphatase PP1γ2 in sperm morphogenesis and epididymal initiation of sperm motility

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Asian Journal of Andrology 2007年第4期9卷 445-452页

作者： Rumela Chakrabarti Lina Cheng Pawan Purl David Soler Srinivasan Vijayaraghavan Department of Biological Sciences Kent State University Kent OH 44242-0001 USA

The serine/threonine phosphatase （PP1） isoform PP1γ2, predominantly expressed in the testis, is a key enzyme in spermatozoa. High PP1γ2 catalytic activity holds motility in check in immature spermatozoa. Inhibition of PP1γ2 causes motility initiation in immature spermatozoa and motility stimulation and changes in flagellar beat parameters in mature spermatozoa. The PP1γ2 isoform is present in all mammalian spermatozoa studied： mouse, rat, hamster, bovine, non-human primate and man. We have now identified at least four of its regulatory proteins that regulate distinct pools of PP1γ2 within spermatozoa. Our studies provide new insights into biochemical mechanisms underlying development and regulation of sperm motility. We hypothesize that changes in sperm PP1γ2 activity as a result of phosphorylation and reversible binding of the regulatory proteins to the catalytic subunit are critical in the development and regulation of motility and the ability of sperm to fertilize eggs. Targeted disruption of the Ppplcc gene, which encodes the PP1γ1 or PP1γ2 isoforms, causes male infertility in mice as a result of impaired spermiogenesis. Our observations suggest that, in addition to motility, the protein phosphatase PP1γ2 might play an isoform-specific function in the development of specialized flagellar structures of mammalian spermatozoa. （Asian J Androl 2007 July; 9： 445--452）

关键词： protein phosphatase epididymis sperm motility spermatogenesis

AMP-activated kinase in human spermatozoa： identification, intracellular localization, and key function in the reRulation of sperm motility

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Asian Journal of Andrology 2017年第6期19卷 707-714页

作者： Violeta Calle-Guisado Ana Hurtado de Llera David Martin-Hidalgo Jose Mijares Maria C Gil Ignacio S Alvarez Maria J Bragado Luis J Garcia-Marin Research Group of Intracellular Signalling and Technology of Reproduction (SINTREP) School of Veterinary Medicine University of Extremadura Caceres Spain Assisted Reproduction Unit at the-Minimally Invasive surgery Center Jesos Uson (CCMIJU) Caceres Spain Norba Clinic Caceres Spain Research Group of Reproduction and Embryo Development (REDES) University of Extremadura Badajoz Spain Extremadura Institute of Assisted Reproduction (IERA) Badajoz Spain.

AMP-activated kinase （AMPK）, a protein that regulates energy balance and metabolism, has recently been identified in boar spermatozoa where regulates key functional sperm processes essential for fertilization. This work＇s aims are AMPK identification, intracellular localization, and their role in human spermatozoa function. Semen was obtained from healthy human donors. sperm AMPK and phospho-Thr172-AMPK were analyzed by Western blotting and indirect immunofluorescence. High- and low-quality sperm populations were separated by a 40%-80% density gradient. Human spermatozoa motility was evaluated by an Integrated Semen Analysis System （ISAS） in the presence or absence of the AMPK inhibitor compound C （CC）. AMPK is localized along the human spermatozoa, at the entire acrosome, midpiece and tail with variable intensity, whereas its active form, phospho-Thr172-AMPK, shows a prominent staining at the acrosome and sperm tail with a weaker staining in the midpiece and the postacrosomal region. Interestingly, spermatozoa bearing an excess residual cytoplasm show strong AMPK staining in this subcellular compartment. Both AMPK and phospho-Thr172-AMPK human spermatozoa contents exhibit important individual variations. Moreover, active AMPK is predominant in the high motility sperm population, where shows a stronger intensity compared with the low motility sperm population. Inhibition of AMPK activity in human spermatozoa by CC treatment leads to a significant reduction in any sperm motility parameter analyzed： percent of motile sperm, sperm velocities, progressivity, and other motility coefficients. This work identifies and points out AMPK as a new molecular mechanism involved in human spermatozoa motility. Further AMPK implications in the clinical efficiency of assisted reproduction and in other reproductive areas need to be studied.

关键词： AMP-activated kinase human spermatozoa immunolocalization sperm motility sperm quality

Relationship between sperm mitochondrial membrane potential,sperm motility,and fertility potential

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Asian Journal of Andrology 2002年第2期4卷 97-103页

作者： TsuyoshiKasai KeigoOgawa 等 DepartmentofObstetricsandGynecology YamanashiMedicalUniversityYamanashi409－3898Japan

OBJECTIVE:To analyze the relationship between sperm mitochondrial membrane potential and sperm motility parameters by means of a computer-assisted sperm analyzer (CASA) and in-vitro fertilization rate(%FR).;METHODS:Semen samples were obtained from 26 men undergoing in vitro fertilization-embryo transfer (IVF-ET). Informed consent was obtained from all men prior to the study. Samples were prepared using wash and swim-up method in HEPES-HTF medium. The sperm motility (%MOT), progressive motility (%PMOT), average path velocity (VAP) microm/s), straight line velocity (VSL) (micro m/s), curvilinear velocity (VCL) (microm/s) and %hyperactivated sperm (%HA), and the %FR were assessed. The samples were incubated in the presence of 2.0 mciromol/L of 5,5',6,6'-tetra-chloro-1,1',3,3'-tetraethylbenzimidazolyl-carbocyanine iodide (JC-1) for 30 min at 37 degrees C in air and washed in PBS before flow cytometry (FACSCalibur: Becton Dickinson) analysis. The mitochondrial probe JC-1 was used to identify the mitochondrial membrane potential. The sperm was divided into three populations according to the fluorescence pattern as follows: the high mitochondrial membrane potential group (n=8), the moderate group (n=5), and the low group (n=13). Statistical analysis was performed using unpaired t-test.;RESULTS:Significant differences were found between the high and the low groups in %MOT (91.1+/-8.5 vs 63.0+/-32.7, mean+/-SD), VAP (73.0+/-14.2 vs 52.1+/-12.5), VCL (127.0+/-28.1 vs 87.0+/-22.6), %HA (27.3+/-23.6 vs 7.2+/-9.0) and %FR [73.2 (48/56) vs 59.0 (69/117)]. No significant differences were found in other CASA parameters.;CONCLUSIONS:When the sperm mitochondrial membrane potential increases, sperm motility parameters and fertility potential will also increase. The JC-1 dye method is useful to predict sperm fertility potential.

关键词： Computer assisted sperm analysis Fertility Fluorescence Hyperactivation In vitro fertilization Membrane potential Mitochondria sperm function sperm motility