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Expression of hepatitis B virus genes in early embryonic cells originated from hamster ova and human spermatozoa transfected with the complete viral genome 被引量:63
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作者 Bahy Ahmed Ali Tian-Hua Huang +1 位作者 Halima-Hassan Salem Qing-Dong Xie 《Asian Journal of Andrology》 SCIE CAS CSCD 2006年第3期273-279,共7页
Aim: To detect the expression of hepatitis B virus (HBV) genes (HB S and C genes) in early embryonic cells after introducing motile human sperm carrying HBV DNA into zona-free hamster oocytes via the in vitro fer... Aim: To detect the expression of hepatitis B virus (HBV) genes (HB S and C genes) in early embryonic cells after introducing motile human sperm carrying HBV DNA into zona-free hamster oocytes via the in vitro fertilization (IVF) technique. Methods: Human sperm-mediated HBV genes were delivered into zona-free hamster oocytes by the IVF method. Polymerase chain reaction (PCR) was used to detect HB S and pre-Core/Core (pre-C/C) coding genes both in one- and two-cell embryos. Reverse transcription-PCR (RT-PCR) analysis was used to study the expression of the two genes. Fluorescence in situ hybridization (FISH) analysis using the full-length HBV DNA as the hybridization probe was performed to confirm the integration of viral DNA in the host embryonic genome. Results: Both HB S and pre-C/C coding genes are present and transcribed in one- and two-cell embryos originated from hamster ova IVF with human spermatozoa carrying HBV DNA sequences. Conclusion: Sperm-mediated HBV genes are able to replicate and express themselves in early embryonic cells. These results provide direct evidence that HBV DNA could transmit vertically to the next generation via the male germ line. 展开更多
关键词 hepatitis B virus gene expression hamster ovary human spermatozoa in vitro fertilization
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Cryopreservation-induced decrease in heat-shock protein 90 in human spermatozoa and its mechanism 被引量:14
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作者 Wen-Lei CAO, Yi-Xin WANG, Zu-Qiong XIANG, Zheng LI Shanghai Institute of Andrology, Renji Hospital, Shanghai Second Medical University, Shanghai 200001, China 《Asian Journal of Andrology》 SCIE CAS CSCD 2003年第1期43-46,共4页
<abstract>Aim: To study the protein changes of spermatozoa associated with sperm motility during sperm cryopreservation and its mechanism. Methods: In 18 healthy men, the seminal sperm motility and HSP90 levels ... <abstract>Aim: To study the protein changes of spermatozoa associated with sperm motility during sperm cryopreservation and its mechanism. Methods: In 18 healthy men, the seminal sperm motility and HSP90 levels were studied before and after cryopreservation using SDS-PAGE, Western blotting and computerized image analysis. Results: The sperm motility declined significantly after cryopreservation (P<0.01). The average grey level and the integrated grey level of sperm HSP90 before cooling were 34.1±3.2 and 243.0±21.6, respectively, while those after thawing were 23.2±2.5 and 105.7±28.5, respectively. Both parameters were decreased significantly (P<0.01). No HSP90 was found in the seminal plasma before and after cryopreservation. Conclusion: HSP90 in human spermatozoa was decreased substantially after cryopreservation. This may result from protein degradation, rather than leakage into the seminal plasma. 展开更多
关键词 human spermatozoa seminal plasma heat-shock proteins 90 western blotting sperm preservation image analysis
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Estimate of oxygen consumption and intracellular zinc concentration of human spermatozoa in relation to motility 被引量:2
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作者 Ralf R.Henkel Kerstin Defosse +2 位作者 Hans-Wilhelm Koyro Norbert Weissmann Wolf-Bernhard Schill 《Asian Journal of Andrology》 SCIE CAS CSCD 2003年第1期3-8,共6页
<abstract>Aim: To investigate the human sperm oxygen/energy consumption and zinc content in relation to motility. Methods: In washed spermatozoa from 67 ejaculates, the oxygen consumption was determined. Followi... <abstract>Aim: To investigate the human sperm oxygen/energy consumption and zinc content in relation to motility. Methods: In washed spermatozoa from 67 ejaculates, the oxygen consumption was determined. Following calculation of the total oxygen consumed by the Ideal Gas Law, the energy consumption of spermatozoa was calculated. In addition, the zinc content of the sperm was determined using an atomic absorption spectrometer. The resulting data were correlated to the vitality and motility. Results: The oxygen consumption averaged 0.24μmol/106 sperm×24 h, 0.28μmol/106 live sperm×24 h and 0.85μmol/106 live & motile sperm×24 h. Further calculations revealed that sperm motility was the most energy consuming process (164.31 mJ/106 motile spermatozoa×24 h), while the oxygen consumption of the total spermatozoa was 46.06 mJ/106 spermatozoa×24 h. The correlation of the oxygen/ energy consumption and zinc content with motility showed significant negative correlations (r= -0.759; P<0.0001 and r=-0.441; P<0.0001, respectively). However, when correlating sperm energy consumption with the zinc content, a significant positive relation (r=0.323; P=0.01) was observed. Conclusion: Poorly motile sperm are actually wasting the available energy. Moreover, our data clearly support the 'Geometric Clutch Model' of the axoneme function and demonstrate the importance of the outer dense fibers for the generation of sperm motility, especially progressive motility. 展开更多
关键词 OXYGEN energy consumption human spermatozoa sperm motility ZINC
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Destabilization of acrosome and elastase influence mediate the release of secretory phospholipase A2 from human spermatozoa 被引量:2
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作者 Jacqueline Leβig Uta Reibetanz +1 位作者 Jiirgen Arnhold Hans-Jtirgen Glander 《Asian Journal of Andrology》 SCIE CAS CSCD 2008年第6期829-836,共8页
Aim: To determine the cellular distribution of secretory phospholipase A2 (sPLA2) in dependence on the acrosomal state and under the action of elastase released under inflammatory processes from leukocytes. Methods... Aim: To determine the cellular distribution of secretory phospholipase A2 (sPLA2) in dependence on the acrosomal state and under the action of elastase released under inflammatory processes from leukocytes. Methods: Acrosome reaction of spermatozoa was triggered by calcimycin. Human leukocyte elastase was used to simulate inflammatory conditions. To visualize the distribution of sPLA2 and to determine the acrosomal state, immunofluorescence techniques and lectin binding combined with confocal laser scanning fluorescence microscopy and flow cytometry were used. Results: Although sPLA2 was detected at the acrosome and tail regions in intact spermatozoa, it disappeared from the head region after triggering the acrosome reaction. This release of sPLA2 was associated with enhanced binding of annexin V-fluoroscein isothiocyanate (FITC) to spermatozoa surfaces, intercalation of ethidium-homodimer I, and binding of FITC-labelled concanavalin A at the acrosomal region. Spermatozoa from healthy subjects treated with elastase were characterized by release of sPLA2, disturbance of acrosome structure, and loss of vitality. Conclusion: The ability of spermatozoa to release secretory phospholipase A2 is related to the acrosomal state. Premature destabi- lization of the acrosome and loss of sPLA2 can occur during silent inflammations in the male genital tract. The distribution pattern of sPLA2 in intact spermatozoa might be an additional parameter for evaluating sperm quality. 展开更多
关键词 acrosome reaction ELASTASE human spermatozoa INFLAMMATION secretory phospholipase A2
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Acetyl-L-carnitine:An Effective Antioxidant against Cryo-damage on Human Spermatozoa with Asthenospermia 被引量:7
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作者 邹宇洁 杨菁 +3 位作者 常硕 徐望明 尹太郎 龙文 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2017年第6期915-921,共7页
A variety of natural and artificial cryoprotectant extenders have been explored to enhance sperm recovery following cryopreservation-thawing process. The current investigation is aimed at evaluating the effect of acet... A variety of natural and artificial cryoprotectant extenders have been explored to enhance sperm recovery following cryopreservation-thawing process. The current investigation is aimed at evaluating the effect of acetyl-L-carnitine on human spermatozoa and reactive species oxygen(ROS) level after freezing-thawing process. The spermatozoa were collected from 35 male patients diagnosed as having asthenospermia. The cryopreservation of human spermatozoa treated with acetyl-L-carnitine at different concentrations(group B: 2.5 mmol/L, group C: 7.5 mmol/L, group D: 15 mmol/L) was compared with control(group A: no acetyl-L-carnitine given). For the frozen-thawed spermatozoa, the viability, motility and DNA integrity were measured by comet assay, acrosome integrity by FITC-PNA staining and ROS level was determined in each group. The results showed that there were no significant differences in motility and viability between group A and group B, while the motility and viability of spermatozoa in group C and group D were significantly increased as compared with those in group A. As compared with group A, the values for DNA integrity parameters including comet rate(CR), tail DNA percentage(TD), tail length(TL) and Oliver tail moment(OTM) were significantly reduced in group C and group D. Group C and group D also displayed a higher proportion of intact acrosome than group A. No significant difference in ROS level was found between group A and group B, while with the increase in acetyl-L-carnitine concentration, the ROS level in groups C and D was significantly reduced as compared with that in group A. In conclusion, acetyl-L-carnitine at a concentration of 7.5 mmol/L is an effective antioxidant against cryo-damage on post-thawed human spermatozoa. 展开更多
关键词 acetyl-L-carnitine human spermatozoa DNA damage acrosome integrity
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Adriamycin induces H2AX phosphorylation in human spermatozoa 被引量:1
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作者 Zhong-Xiang Li Ting-Ting Wang +4 位作者 Yan-Ting Wu Chen-Ming Xu Min-Yue Dong Jian-Zhong Sheng He-Feng Huang 《Asian Journal of Andrology》 SCIE CAS CSCD 2008年第5期749-757,共9页
Aim: To investigate whether adriamycin induces DNA damage and the formation of γH2AX (the phosphorylated form of histone H2AX) foci in mature spermatozoa. Methods: Human spermatozoa were treated with adriamycin a... Aim: To investigate whether adriamycin induces DNA damage and the formation of γH2AX (the phosphorylated form of histone H2AX) foci in mature spermatozoa. Methods: Human spermatozoa were treated with adriamycin at different concentrations, γH2AX was analyzed by immunofluorescent staining and flow cytometry and doublestrand breaks (DSB) were detected by the comet assay. Results: The neutral comet assay revealed that the treatment with adriamycin at 2 μg/mL for different times (0.5, 2, 8 and 24 h), or for 8 h at different concentrations (0,4, 2 and 10 μg/mL), induced significant DSB in spermatozoa. Immunofluorent staining and flow cytometry showed that the expression of γH2AX was increased in a dose-dependent and time-dependant manner after the treatment of adriamycin. Adriamycin also induced the concurrent appearance of DNA maintenance/repair proteins RAD50 and 53BP 1 with γH2AX in spermatozoa. Wortmannin, an inhibitor of the phosphatidylinositol 3-kinase (PI3K) family, abolished the co-appearance of these two proteins with γH2AX. Conclusion: Human mature spermatozoa have the same response to DSB-induced H2AX phosphorylation and subsequent recruitment of DNA maintenance/ repair proteins as somatic cells. 展开更多
关键词 ADRIAMYCIN human spermatozoa DNA double strand-breaks γH2AX
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The expression and significance of CATSPER1 in human testis and ejaculated spermatozoa 被引量:18
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作者 Hong-Gang Li Ai-Hua Liao Xiao-Fang Ding Hui Zhou Cheng-Liang Xiong 《Asian Journal of Andrology》 SCIE CAS CSCD 2006年第3期301-306,共6页
Aim: To investigate the distribution of cation channel of sperm 1 (CATSPER1) protein and the presence of CATSPER1 mRNA in human testis and ejaculated spermatozoa. The influence of anti-human CATSPER1 antibody upon ... Aim: To investigate the distribution of cation channel of sperm 1 (CATSPER1) protein and the presence of CATSPER1 mRNA in human testis and ejaculated spermatozoa. The influence of anti-human CATSPER1 antibody upon human sperm motility was used to evaluate the function of human CATSPER1 and to estimate its possible use as a target for immunocontraception. Methods: Human ejaculated sperm from normozoospermic donors (n = 12) and liquid nitrogen frozen human testis were used for the study of mRNA and protein expression of CATSPER1 by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry, respectively. Spermatozoa from normozoospermic donors (n = 12) were individually processed using a swim-up procedure and were then incubated with CATSPER1 antibody at final concentrations of 20, 4 and 0.8 μg/mL. After 1, 2 and 6 h incubation, progressive motility and fast progressive motility were measured by means of computer-assisted semen analysis. Results: CATSPER1 transcript was detected in both human testis and each human ejaculated semen sample. CATSPER1 protein expressed in the membrane of spermatid and was localized in the principal piece of the sperm tail. The application of CATSPER1 antibody at all concentrations significantly inhibited both progressive motility and fast progressive motility after 1, 2 and 6 h incubation, and significant dose-dependent changes were observed. Conclusion: CATSPER1 is meiotically and post-meiotically expressed in human testis tissue. CATSPER1 mRNA in human ejaculated spermatozoa could be a more feasible target for study and infertility screening than testis biopsy. In addition, our results suggest that human CATSPER1 could be a possible target for immunocontraception. 展开更多
关键词 cation channel of sperm 1 TESTIS spermatozoa MOTILITY IMMUNOCONTRACEPTION human
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Shortening of alkaline DNA unwinding time does not interfere with detecting DNA damage to mouse and human spermatozoa in the comet assay 被引量:1
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作者 Hirokazu Kusakabe Hiroyuki Tateno 《Asian Journal of Andrology》 SCIE CAS CSCD 2011年第1期172-174,共3页
The comet assay was performed on mouse and human spermatozoa to examine the effect of alkaline DNA unwinding time. The spermatozoa were treated in vitrowith the DNA-damaging agents, methyl methanesulfonate (MMS) or ... The comet assay was performed on mouse and human spermatozoa to examine the effect of alkaline DNA unwinding time. The spermatozoa were treated in vitrowith the DNA-damaging agents, methyl methanesulfonate (MMS) or hydrogen peroxide (Hz02), and then embedded in agarose gel on glass sl ides. The slides were immersed in alkaline solution (〉pH 13) for 1, 5, 10 and 20 min, and then subjected to the electrophoresis under neutral conditions. In mouse spermatozoa, comet tails seen in solvent controls became brighter and longer as the alkaline DNA unwinding time increased. However, in the MMS-treated mouse spermatozoa, a smaller difference in the damage from that in the solvent control was seen with time within a dose. DNA damage induced by H2O2 could also be detected accurately after alkali treatment for 1-20 min. In human spermatozoa, DNA damage induced by MMS and H2O2 could be detected in a dose-dependent manner after alkali treatment for 1 min. The ability of the comet assay to detect DNA damage was not adversely affected by the short period (1 min) of the alkaline DNA unwinding time. 展开更多
关键词 ALKALINE comet assay DNA unwinding human mice spermatozoa
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The expression of vitronectin,a ligand of integrin,on human spermatozoa and its role in fertilization
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作者 Bronson Rictard 《Asian Journal of Andrology》 SCIE CAS CSCD 2001年第1期74-74,共1页
Objective: To study the vitronectin expression in human spermato-zoa and its role in fertilization. Methods: Spermatozoa from 14 fer-tile and 8 infertile men with normal semen data were studied. Follow-ing recovery of... Objective: To study the vitronectin expression in human spermato-zoa and its role in fertilization. Methods: Spermatozoa from 14 fer-tile and 8 infertile men with normal semen data were studied. Follow-ing recovery of motile populations by swim-up, spermatozoa were ca-pacitated and immunostained with rabbit anti-human Vn polyclonal an-tibody and goat anti-rabbit IgG-FTTC second antibody. The percentageof spermatozoa expressing Vn was determined using a FAScan flowcytometer. Meanwhile, the fertilizing ability of capacitated spermato-zoa was determined with human spermatozoa zona-free hamster eggpenetration assay (SPA). Results: The mean ± s proportion ofspermatozoa expressing Vn of fertile men was 21.24% ± 11.70% and3.64±3.27% for infertile men (P<0.05). The penetration rate ofSPA in the fertile group was > 10%, but that in the infertile group ,< 10%. There is a correlation between positive sperm Vn expressionand percentage of eggs penetrated (r=0.476). Conclusion: Theseresults indicate the expression of Vn in human capacitated spermatozoaand its correlation with the fertilizing ability. The abnormal expressionof Vn on human spermatozoa may be one of the unexplained infertilereasons. (Reprod Contracep 2001; 21: 24-28) 展开更多
关键词 vitronectin (Vn) human spermatozoa FERTILIZATION
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Expression of hepatitis B virus genes in early embryonic cells originated from hamster ova and human spermatozoa transfected with the complete viral genome 被引量:5
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作者 BabyAhmedAli Halima-HassanSalem 《Asian Journal of Andrology》 SCIE CAS CSCD 2006年第A03期273-279,385,共5页
Aim:To detect the expression of hepatitis B virus(HBV)genes(HB S and C genes)in early embryonic cells after introducing motile human sperm carrying HBV DNA into zona-free hamster oocytes via the in vitro fertilization... Aim:To detect the expression of hepatitis B virus(HBV)genes(HB S and C genes)in early embryonic cells after introducing motile human sperm carrying HBV DNA into zona-free hamster oocytes via the in vitro fertilization(IVF) technique.Methods:Human sperm-mediated HBV genes were delivered into zona-free hamster oocytes by the IVF method.Polymerase chain reaction(PCR)was used to detect HB S and pre-Core/Core(pre-C/C)coding genes both in one-and two-cell embryos.Reverse transcription-PCR(RT-PCR)analysis was used to study the expression of the two genes.Fluorescence in situ hybridization(FISH)analysis using the full-length HBV DNA as the hybridization probe was performed to confirm the integration of viral DNA in the host embryonic genome.Results:Both HB S and pre-C/C coding genes are present and transcribed in one-and two-cell embryos originated from hamster ova IVF with human spermatozoa carrying HBV DNA sequences.Conclusion:Sperm-mediated HBV genes are able to replicate and express themselves in early embryonic cells.These results provide direct evidence that HBV DNA could transmit vertically to the next generation via the male germ line. 展开更多
关键词 hepatitis B virus gene expression hamster ovary human spermatozoa in vitro fertilization
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The expression and significance of CATSPER1 in human testis and ejaculated spermatozoa
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作者 Hong-Gang Li Ai-Hua Liao +2 位作者 Xiao-Fang Ding Hui Zhou Cheng-Liang Xiong 《Asian Journal of Andrology》 SCIE CAS CSCD 2006年第A03期301-306,386,共5页
Aim:To investigate the distribution of cation channel of sperm 1(CATSPER1)protein and the presence of CATSPER1 mRNA in human testis and ejaculated spermatozoa.The influence of anti-human CATSPER1 antibody upon human s... Aim:To investigate the distribution of cation channel of sperm 1(CATSPER1)protein and the presence of CATSPER1 mRNA in human testis and ejaculated spermatozoa.The influence of anti-human CATSPER1 antibody upon human sperm motility was used to evaluate the function of human CATSPER1 and to estimate its possible use as a target for immunocontraception.Methods:Human ejaculated sperm from normozoospermic donors(n=12)and liquid nitro- gen frozen human testis were used for the study of mRNA and protein expression of CATSPER1 by reverse transcliption polymerase chain reaction(RT-PCR)and immunohistochemistry,respectively.Spermatozoa from nonnozoospermic donors(n=12)were individually processed using a swim-up procedure and were then incubated with CATSPER 1 antibody at final concentrations of 20,4 and 0.8μg/mL.After 1,2 and 6h incubation,progressive motility and fast progressive motility were measured by means of computer-assisted semen analysis.Results:CATSPER1 transcript was detected in both human testis and each human ejaculated semen sample.CATSPER1 protein expressed in the membrane of spermatid and was localized in the principal piece of the sperm tail.The application of CATSPER1 antibody at all concentrations significantly inhibited both progressive motility and fast progressive motility after 1,2 and 6h incubation,and significant dose-dependent changes were observed.Conclusion:CATSPER1 is meiotically and post-meiotically expressed in human testis tissue.CATSPER1 mRNA in human ejaculated spermatozoa could be a more feasible target for study and infertility screening than testis biopsy.In addition,our results suggest that human CATSPER1 could be a possible target for immunocontraception.(Asian J Androl 2006 May:8:301-306) 展开更多
关键词 (?)ation channel of sperm 1 TESTIS spermatozoa MOTILITY IMMUNOCONTRACEPTION human
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Human sperm pattern of movement during chemotactic 'e-orientation towards a progesterone source 被引量:2
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作者 Cecilia Soledad Blengini Maria Eugenia Teves Diego Rafael Ufiates Hetor Alejandro Guidobaldi LauraVirginia Gatica Laura Cecilia Giojalas 《Asian Journal of Andrology》 SCIE CAS CSCD 2011年第5期769-773,共5页
Human spermatozoa may chemotactically find out the egg by following an increasing gradient of attractant molecules. Although human spermatozoa have been observed to show several of the physiological characteristics of... Human spermatozoa may chemotactically find out the egg by following an increasing gradient of attractant molecules. Although human spermatozoa have been observed to show several of the physiological characteristics of chemotaxis, the chemotactic pattern of movement has not been easy to describe. However, it is apparent that chemotactic cells may be identified while returning to the attractant source. This study characterizes the pattern of movement of human spermatozoa during chemotactic re-orientation towards a progesterone source, which is a physiological attractant candidate. By means of videomicroscopy and image analysis, a chemotactic pattern of movement was identified as the spermatozoon returned towards the source of a chemotactic concentration of progesterone (10 pmol I^-1). First, as a continuation of its original path, the spermatozoon swims away from the progesterone source with linear movement and then turns back with a transitional movement that can be characterized by an increased velocity and decreased linearity. This sperm behaviour may help the spermatozoon to re-orient itself towards a progesterone source and may be used to identify the few cells that are undergoing chemotaxis at a given time. 展开更多
关键词 CHEMOTAXIS human spermatozoa PROGESTERONE
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Studies on the integration of hepatitis B virus DNA sequence in human sperm chromosomes 被引量:49
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作者 Jian-Min HUANG Tian-Hua HUANG +3 位作者 Huan-Ying QIU Xiao-Wu FANG Tian-Gang ZHUANG Jie-Wen QIU 《Asian Journal of Andrology》 SCIE CAS CSCD 2002年第3期209-212,共4页
Aim: To study the integration of hepatitis B virus (HBV) DNA into sperm chromosomes in hepatitis B patients and the features of its integration. Methods: Sperm chromosomes of 14 subjects (5 healthy controls and 9 HB p... Aim: To study the integration of hepatitis B virus (HBV) DNA into sperm chromosomes in hepatitis B patients and the features of its integration. Methods: Sperm chromosomes of 14 subjects (5 healthy controls and 9 HB patients, including 1 acute hepatitis B, 2 chronic active hepatitis B, 4 chronic persistent hepatitis B, 2 HBsAg chronic carriers with no clinical symptoms) were prepared using interspecific in vitro fertilization between zona-free hamster oocytes and human spermatozoa. Fluorescence in situ hybridization (FISH) to sperm chromosome spreads was carried out with biotin-labeled full length HBV DNA probe to detect the specific HBV DNA sequences in the sperm chromosomes. Results: Specific fluorescent signal spots for HBV DNA were seen in sperm chromosomes of one patient with chronic persistent hepatitis B. In 9(9/42) sperm chromosome complements containing fluorescent signal spots, one presented 5 obvious FISH spots and the others 2 to 4 signals. The fluorescence intensity showed significant difference among the signal spots. The distribution of signal sites among chromosomes seems to be random. Conclusion: HBV could integrate into human sperm chromosomes. Results suggest that the possibility of vertical transmission of HBV via the germ line to the next generation is present. 展开更多
关键词 hepatitis B virus spermatozoa human chromosomes fluorescence in situ hybridization virus integration vertical disease transmission
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Protein tyrosine phosphorylation of the human sperm head during capacitation: immunolocalization and relationship with acquisition of sperm-fertilizing ability 被引量:1
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作者 Arcangelo Barbonetti Maria Rosaria C. Vassallo +6 位作者 Giuliana Cordeschi Dimitrios Venetis Andrea Carboni Alessandra Sperandio Giorgio Felzani Sandro Francavilla Felice Francavilla 《Asian Journal of Andrology》 SCIE CAS CSCD 2010年第6期853-861,共9页
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 abi... 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. 展开更多
关键词 acrosome reaction CAPACITATION human spermatozoa sperm-oocyte fusion tyrosine phosphorylation
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不同冷冻保护剂对人精子染色体及超微结构影响的研究 被引量:6
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作者 张巧玉 常青 +2 位作者 史常旭 罗平 蔡敏 《重庆医学》 CAS CSCD 2002年第7期550-552,共3页
目的 旨在了解不同冷冻保护剂 (CPM)对人精子染色体畸变率及精子超微结构相对正常精子百分率的影响。方法 应用甘油、甘油 卵黄 柠檬酸钠 (GYC)及甘油 卵黄 柠檬酸钠 L 谷氨酰胺 (GYCG)三种冷冻保护剂 ,检测冻贮前后精子染色体畸变... 目的 旨在了解不同冷冻保护剂 (CPM)对人精子染色体畸变率及精子超微结构相对正常精子百分率的影响。方法 应用甘油、甘油 卵黄 柠檬酸钠 (GYC)及甘油 卵黄 柠檬酸钠 L 谷氨酰胺 (GYCG)三种冷冻保护剂 ,检测冻贮前后精子染色体畸变率和超微结构相对正常的精子百分率。结果  (1)精子染色体畸变率及精子性染色体的比例在三种冷冻保护剂冷冻前后均无显著改变 ;(2 )在三种冷冻保护剂组冷冻复温后 ,精子超微结构相对正常的精子百分率显著下降 ,但GYCG组高于甘油、GYC组。结论 本研究中三种冷冻保护剂不会增加精子染色体畸变率 ,也不会引起精子性染色体比例失衡 。 展开更多
关键词 冷冻保护剂 人精子 染色体 超微结构 影响 研究
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人精子中芳香化酶表达与精子功能的关系 被引量:8
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作者 郑洁 何亚平 +3 位作者 张金虎 何彦芳 高小平 岳利民 《生殖与避孕》 CAS CSCD 北大核心 2005年第5期284-289,共6页
目的:研究精子细胞色素P450芳香化酶(P450arom)的表达及其与精子功能状态和受精力的关系。方法:以人精子穿透去透明带金黄地鼠卵异种体外受精试验(SPA)检测精子受精力;以三色法染色观察精子顶体反应(AR)的发生率。采用RT-PCR,用P450arom... 目的:研究精子细胞色素P450芳香化酶(P450arom)的表达及其与精子功能状态和受精力的关系。方法:以人精子穿透去透明带金黄地鼠卵异种体外受精试验(SPA)检测精子受精力;以三色法染色观察精子顶体反应(AR)的发生率。采用RT-PCR,用P450arom/GAPDH光密度值的比值代表P450arom的表达水平。结果:精子P450arom表达水平与受精率有一定的相关性(生育组r=0.5622;不育组r=0.6071)。正常男性与不明原因不育症患者精子P450arom/GAPDH比值分别为0.60±0.29,0.39±0.16,有显著差异(P<0.02)。P450arom表达水平与精子AR的发生率也有一定相关性(生育组r=0.5817;不育组r=0.5535)。结论:人精子中存在着P450arom表达产物;P450arom可能与精子功能有一定关系;P450arom表达异常可能与一些不明原因不育有关。 展开更多
关键词 芳香化酶 人精子 受精 MRNA
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17β-雌二醇诱导人精子顶体反应及胞内钙离子增加的研究 被引量:5
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作者 何彦芳 岳利民 +3 位作者 何亚平 张金虎 郑洁 高小平 《四川大学学报(医学版)》 CAS CSCD 北大核心 2005年第4期500-502,共3页
目的探讨雌激素对人精子的激活作用及其可能的作用机制。方法以17β-雌二醇(E2)及非透膜性大分子雌二醇-牛血清白蛋白交联物(E2-BSA)分别作用于生育力正常男性的精子,以三色法染色评价精子顶体反应(AR)发生率,流式细胞术检测精子内游离C... 目的探讨雌激素对人精子的激活作用及其可能的作用机制。方法以17β-雌二醇(E2)及非透膜性大分子雌二醇-牛血清白蛋白交联物(E2-BSA)分别作用于生育力正常男性的精子,以三色法染色评价精子顶体反应(AR)发生率,流式细胞术检测精子内游离Ca2+浓度[Ca2+]i的变化。结果E2可引起获能精子AR率明显增加,使[Ca2+]i快速升高,对非获能精子则无明显影响;E2-BSA同样可以提高获能精子AR率及[Ca2+]i,其作用与E2相似;E2诱导获能精子[Ca2+]i升高依赖于胞外Ca2+的内流。结论雌激素对人精子有一定的激活作用,该作用可能是通过与人精子膜上的雌激素结合位点作用后使胞外Ca2+内流而实现的。 展开更多
关键词 雌激素 人精子 顶体反应 钙离子
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考马斯亮蓝染色法检测人精子顶体反应的评价 被引量:22
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作者 江一平 卓丹心 +2 位作者 陈勇 林芸秀 韩咪莎 《解剖学报》 CAS CSCD 北大核心 1998年第3期332-336,共5页
为建立一种检测人精子顶体反应的简便实用的新技术,用3.5%高氯酸水溶液配制0.05%考马斯亮蓝(R250)染色液浸染人精子30min,顶体完整者顶体区染成紫蓝色,顶体反应者则不染。对获能前后和钙离子载体A23187诱... 为建立一种检测人精子顶体反应的简便实用的新技术,用3.5%高氯酸水溶液配制0.05%考马斯亮蓝(R250)染色液浸染人精子30min,顶体完整者顶体区染成紫蓝色,顶体反应者则不染。对获能前后和钙离子载体A23187诱导顶体反应的精子进行染色并与经典的顶体反应检测技术PSA法对照,两种方法检测顶体反应率无显著差异。方法学检验证明新技术结果可靠。本法操作简便、经济,普通显微镜即可检测,易于推广,克服了原有技术复杂昂贵的缺点,具有研究和临床诊断的良好价值。 展开更多
关键词 精子 顶体反应 考马斯亮蓝 染色法
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海藻糖和蛋黄在人类精子冻干保存中的保护作用 被引量:5
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作者 舒志全 张浩波 +7 位作者 张宁 俞斐 李晶 何立群 陈子江 季维智 杨世华 高大勇 《中国生物医学工程学报》 CAS CSCD 北大核心 2007年第1期121-125,共5页
人类精子冻干保存中损伤除了来自环境溶液对精子生理活性的损伤外,还会来自于冷冻和干燥过程。目前精子冻干保存中保护剂选取基本上都从生理化学角度考虑,目的为抑制精子内酶活性,没有考虑对冷冻和干燥损伤的抑制。大量文献表明蛋黄能... 人类精子冻干保存中损伤除了来自环境溶液对精子生理活性的损伤外,还会来自于冷冻和干燥过程。目前精子冻干保存中保护剂选取基本上都从生理化学角度考虑,目的为抑制精子内酶活性,没有考虑对冷冻和干燥损伤的抑制。大量文献表明蛋黄能减小冷冻对精子的损伤,而二糖(特别是海藻糖)在细胞干燥中具有特殊保护作用。本研究在前人一般使用的精子冻干保护剂中加入海藻糖,或同时加入海藻糖和蛋黄,通过伊红Y染色、低渗膨胀和电镜显微观测实验,比较不同组成的保护剂对精子结构和膜的保护作用。结果表明,通过加入海藻糖修正的保护剂能提高对精子结构的保护效果,当同时加入海藻糖和蛋黄时效果更好。 展开更多
关键词 人类精子 冻干 保护剂 海藻糖 蛋黄
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活性氧致人精子运动功能和存活力变化的分析 被引量:25
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作者 商学军 叶章群 +2 位作者 熊承良 黄东晖 王咏梅 《中华男科学杂志》 CAS CSCD 2003年第6期439-442,共4页
目的 :对活性氧 (ROS)作用后精子的运动参数和存活率的变化进行分析 ,以证实ROS是否为引起精子运动功能障碍的病因之一。 方法 :采用Percoll梯度离心法选择具有正常生理功能的人精子作为正常精子模型 ,应用次黄嘌呤 黄嘌呤氧化酶体系产... 目的 :对活性氧 (ROS)作用后精子的运动参数和存活率的变化进行分析 ,以证实ROS是否为引起精子运动功能障碍的病因之一。 方法 :采用Percoll梯度离心法选择具有正常生理功能的人精子作为正常精子模型 ,应用次黄嘌呤 黄嘌呤氧化酶体系产生ROS ,在有氧环境下与精子模型孵育后 ,运用计算机辅助精液分析 (CASA)系统 ,分析ROS攻击后精子运动参数的改变。 结果 :与对照组相比 ,正常精子模型与ROS孵育 30min后 ,精子活动率、曲线速度 (VCL)、直线速度 (VSL)、平均路径速度 (VAP)均明显下降 (P <0 .0 0 1) ,头侧摆幅度 (ALH)尚无明显改变 (P >0 .0 5 ) ,而与ROS孵育 6 0min后 ,精子运动功能近乎丧失 ,精子主要运动参数趋向于零。 结论 :ROS与正常精子作用后 ,可导致精子运动功能下降 ,从而证实ROS为精子运动功能障碍的病因之一。 展开更多
关键词 活性氧 运动参数 存活力 精子
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