目的研究常用有机溶剂对正常男性精子体外运动活力的影响。方法取正常男性精子,采用上游优化法处理,制备成精子悬液并分成4组,前3组分别加入二甲亚砜,使二甲亚砜终浓度分别为0.5%,1%和2%,pH值均为7.5,渗透压为290 mosm.(kg.H2O-1),D组...目的研究常用有机溶剂对正常男性精子体外运动活力的影响。方法取正常男性精子,采用上游优化法处理,制备成精子悬液并分成4组,前3组分别加入二甲亚砜,使二甲亚砜终浓度分别为0.5%,1%和2%,pH值均为7.5,渗透压为290 mosm.(kg.H2O-1),D组加入等量Ham’s F10培养液作对照。分别于孵育15,30,45和60 m in后采用CASA进行分析。参照上述方法,考察有机溶剂甘油、聚乙二醇、吐温80对精子体外运动活力的影响。甘油终浓度分别调至1%,5%和10%,聚乙二醇终浓度分别调至1%,5%和10%,吐温80终浓度分别调至1%,5%和10%。结果2%二甲亚砜作用于精子60 m in能导致精子全部死亡,10%甘油作用60 m in后对精子运动参数抑制率达73.8%,10%聚乙二醇400作用于人精子60 m in可致其全部死亡,10%吐温80作用60 m in后对精子运动参数抑制率达95.5%。加入不同浓度二甲亚砜、甘油、聚乙二醇400和吐温80的各组前向运动精子百分率、曲线运动速度、直线运动速度和平均路径速度均较空白对照组显著降低(均P<0.05)。结论高浓度有机溶剂体外可抑制正常男性精子存活率和运动活力。展开更多
Perfluorooctane sulfonate (PFOS) has emerged as one of the most concerning contaminants in recent years. This study aimed to investigate the acute toxicity effect of PFOS on sperm viability, kinematics and fertiliza...Perfluorooctane sulfonate (PFOS) has emerged as one of the most concerning contaminants in recent years. This study aimed to investigate the acute toxicity effect of PFOS on sperm viability, kinematics and fertilization success in zebrafish (Danio rerio). Sperm were activated in aqueous media containing a range of PFOS concentrations (0, 0.09, 0.9 and 9 mg/L). Viabilities and kinematics of the sperm exposed to different PFOS treatments were assessed via computer-assisted sperm analysis (CASA) at 20, 40, 60, and 80 s after activation. PFOS exposure decreased the percentage of motile sperm, the curvilinear velocity (VCL), and the mean angular displacement (MAD) of spermatozoa, but showed no influence on the straight- line velocity (VSL) or the angular path velocity (VAP). Furthermore, a significant decrease in fertilization success was observed in spermatozoa that were exposed to 0.9 mg/L PFOS or more. These findings indicate that PFOS pollution in natural aquatic environment may be a potential threaten to successful reproduction of fish.展开更多
A total of 18 ejaculates were collected, once per week, from six fertile stallions for three consecutive weeks in October and November, to compare motility over time between extenders using four semen processing treat...A total of 18 ejaculates were collected, once per week, from six fertile stallions for three consecutive weeks in October and November, to compare motility over time between extenders using four semen processing treatments. Four total aliquots of semen were used. Two aliquots of each semen collection were extended in either INRA96 or an experimental proprietary milk-based extender Walworth (WW) extender, and each was designed for multi-day storage of fresh chilled semen. Each aliquot was divided again, and either centrifuged at 600 μg for 10 min without cushion, or not centrifuged and extended to a final concentration of 25 × 10^6 spermatozoa/mL. The treatments evaluated were INRA96 without centrifugation (INRA-NC) or with centrifugation (INRA-C), and Walworth extender without centrifugation (WW-NC) or with centrifugation (WW-C). Total and progressive motility were measured using Sperm Vision~ CASA at 0, 24, 48 and 72 h post-collection. Samples were stored at 4 ℃. No differences were found between extenders in progressive (P = 0.13) or total motility (P = 0.14) over the four different time points without centrifugation. However, ejaculates processed in INRA-C group had the greater total and progressive motility (P 〈 0.05) over the four time points than ejaculates in the WW-C group. It was found that centrifugation and re-suspension of stallion semen in INRA96 improved the longevity of fresh chilled semen. However, when not using centrifugation, the Walworth extender proved to be as effective for maintaining spermatozoa motility across all time points as 1NRA96 , and may be an alternative for use in the equine breeding industry.展开更多
The present study evaluated the ef fects of chilled storage and cryopreservation on sperm motion characteristics, antioxidant enzyme activities, and lipid peroxidation in the Pacific cod Gadus macrocephalus. Sperm mot...The present study evaluated the ef fects of chilled storage and cryopreservation on sperm motion characteristics, antioxidant enzyme activities, and lipid peroxidation in the Pacific cod Gadus macrocephalus. Sperm motility and the activities of superoxide dismutase(SOD), catalase(CAT), glutathione peroxidase(GPx), glutathione reductase(Gr), and lipid peroxidation(measured via malondialdehyde(MDA) content) were determined after the milt was stored at 4°C for 12 h, cryopreserved without cryoprotectant in 12% propylene glycol(PG), cryopreserved in 12% PG+0.1 mol/L trehalose, or cryopreserved in 12% PG spermatozoa but centrifuged to decant the supernatant prior to cryopreservation(only sperm cells were cryopreserved). After chilled storage or cryopreservation, the SOD, CAT and GPx activities were reduced in sperm cells and increased in seminal plasma in almost all treatments; sperm motility parameters were also decreased. However, the addition of trehalose into the cryoprotectant could signifi cantly improve the postthaw sperm quality as revealed by the sperm average path velocity. This improvement might be attributed to the function of trehalose in scavenging reactive oxygen species. Chilled storage and cryopreservation had signifi cant eff ects on sperm motion characteristics, antioxidant enzyme activities, and lipid peroxidation in the Pacific cod.展开更多
文摘目的研究常用有机溶剂对正常男性精子体外运动活力的影响。方法取正常男性精子,采用上游优化法处理,制备成精子悬液并分成4组,前3组分别加入二甲亚砜,使二甲亚砜终浓度分别为0.5%,1%和2%,pH值均为7.5,渗透压为290 mosm.(kg.H2O-1),D组加入等量Ham’s F10培养液作对照。分别于孵育15,30,45和60 m in后采用CASA进行分析。参照上述方法,考察有机溶剂甘油、聚乙二醇、吐温80对精子体外运动活力的影响。甘油终浓度分别调至1%,5%和10%,聚乙二醇终浓度分别调至1%,5%和10%,吐温80终浓度分别调至1%,5%和10%。结果2%二甲亚砜作用于精子60 m in能导致精子全部死亡,10%甘油作用60 m in后对精子运动参数抑制率达73.8%,10%聚乙二醇400作用于人精子60 m in可致其全部死亡,10%吐温80作用60 m in后对精子运动参数抑制率达95.5%。加入不同浓度二甲亚砜、甘油、聚乙二醇400和吐温80的各组前向运动精子百分率、曲线运动速度、直线运动速度和平均路径速度均较空白对照组显著降低(均P<0.05)。结论高浓度有机溶剂体外可抑制正常男性精子存活率和运动活力。
基金Supported by the National Natural Science Foundation of China(Nos.40632009,31300340)the Natural Science Foundation Project of Chongqing(No.cstc2015jcyjA20016)the Youth Top-notch Talent Cultivation Program of Chongqing Normal University(No.14CSBJ08)
文摘Perfluorooctane sulfonate (PFOS) has emerged as one of the most concerning contaminants in recent years. This study aimed to investigate the acute toxicity effect of PFOS on sperm viability, kinematics and fertilization success in zebrafish (Danio rerio). Sperm were activated in aqueous media containing a range of PFOS concentrations (0, 0.09, 0.9 and 9 mg/L). Viabilities and kinematics of the sperm exposed to different PFOS treatments were assessed via computer-assisted sperm analysis (CASA) at 20, 40, 60, and 80 s after activation. PFOS exposure decreased the percentage of motile sperm, the curvilinear velocity (VCL), and the mean angular displacement (MAD) of spermatozoa, but showed no influence on the straight- line velocity (VSL) or the angular path velocity (VAP). Furthermore, a significant decrease in fertilization success was observed in spermatozoa that were exposed to 0.9 mg/L PFOS or more. These findings indicate that PFOS pollution in natural aquatic environment may be a potential threaten to successful reproduction of fish.
文摘A total of 18 ejaculates were collected, once per week, from six fertile stallions for three consecutive weeks in October and November, to compare motility over time between extenders using four semen processing treatments. Four total aliquots of semen were used. Two aliquots of each semen collection were extended in either INRA96 or an experimental proprietary milk-based extender Walworth (WW) extender, and each was designed for multi-day storage of fresh chilled semen. Each aliquot was divided again, and either centrifuged at 600 μg for 10 min without cushion, or not centrifuged and extended to a final concentration of 25 × 10^6 spermatozoa/mL. The treatments evaluated were INRA96 without centrifugation (INRA-NC) or with centrifugation (INRA-C), and Walworth extender without centrifugation (WW-NC) or with centrifugation (WW-C). Total and progressive motility were measured using Sperm Vision~ CASA at 0, 24, 48 and 72 h post-collection. Samples were stored at 4 ℃. No differences were found between extenders in progressive (P = 0.13) or total motility (P = 0.14) over the four different time points without centrifugation. However, ejaculates processed in INRA-C group had the greater total and progressive motility (P 〈 0.05) over the four time points than ejaculates in the WW-C group. It was found that centrifugation and re-suspension of stallion semen in INRA96 improved the longevity of fresh chilled semen. However, when not using centrifugation, the Walworth extender proved to be as effective for maintaining spermatozoa motility across all time points as 1NRA96 , and may be an alternative for use in the equine breeding industry.
基金Supported by the National Natural Science Foundation of China(Nos.31072212,41076100)the CAS Scientific and Technological Innovation Program for Cross and Cooperative Team,Marine Economy Innovative Development Project(No.12PYY001SF08)the National Key Basic Program of Science and Technology Platforms of Aquaculture Stock Resources,Shandong Technology Development Project(No.2013GHY11509)
文摘The present study evaluated the ef fects of chilled storage and cryopreservation on sperm motion characteristics, antioxidant enzyme activities, and lipid peroxidation in the Pacific cod Gadus macrocephalus. Sperm motility and the activities of superoxide dismutase(SOD), catalase(CAT), glutathione peroxidase(GPx), glutathione reductase(Gr), and lipid peroxidation(measured via malondialdehyde(MDA) content) were determined after the milt was stored at 4°C for 12 h, cryopreserved without cryoprotectant in 12% propylene glycol(PG), cryopreserved in 12% PG+0.1 mol/L trehalose, or cryopreserved in 12% PG spermatozoa but centrifuged to decant the supernatant prior to cryopreservation(only sperm cells were cryopreserved). After chilled storage or cryopreservation, the SOD, CAT and GPx activities were reduced in sperm cells and increased in seminal plasma in almost all treatments; sperm motility parameters were also decreased. However, the addition of trehalose into the cryoprotectant could signifi cantly improve the postthaw sperm quality as revealed by the sperm average path velocity. This improvement might be attributed to the function of trehalose in scavenging reactive oxygen species. Chilled storage and cryopreservation had signifi cant eff ects on sperm motion characteristics, antioxidant enzyme activities, and lipid peroxidation in the Pacific cod.