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1.
研究了超低温冷冻保存(-196℃)对脊尾白虾精子内琥珀酸脱氢酶(SDH)、乳酸脱氢酶(LDH)、Na+/K+-ATP酶、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽还原酶(GR)与顶体酶活性的影响,以期为提高脊尾白虾精子超低温冷冻效果提供理论依据.设置对照组(未添加抗冻剂)、3个实验组[分别添加DMSO (V/V) 10.0％、12.5％、15.0％],于冷冻0、1、3、5、7、15d取样测定各酶活性.结果表明,经冷冻后,除GR外,其他所测酶活性均出现显著下降(P＜0.05),且以对照组酶活性下降幅度最大.GR活性在冷冻7d内显著升高,且对照组明显高于实验组(P＜0.05),在冷冻15d时又出现下降.添加15.0％ DMSO组所测酶活性均高于同期其它各组,表明15.0％ DMSO对精子内酶的保护作用较好.冷冻15 d后15.0％ DMSO组的SDH、LDH和Na+/K+-ATP酶活性由(28.500±1.453) U/mL、(1290.836±27.603) U/L和(2.605-0.232) μmol/(mg·h)分别降至(15.300±0.950) U/mL、(363.713-13.943) U/L和(0.542-0.186) μmol/(mg.h);SOD和CAT活性由(106.497±7.217) U/mL、(383.632±4.731)U/g分别降至(17.036 ±0.321)U/mL、(166.940±1.910) U/g;顶体酶活性从(3.521±0.010)μIU/106降至(1.212±0.043)μIU/106;而GR活性由(217.042±6.962) U/L上升至(302.787±24.558)U/L.从冷冻后各酶下降幅度来看,超低温冷冻对SOD活性的影响最大,其次是Na+/K+-ATP酶. 相似文献
2.
为了解抗冻剂对施氏鲟(Acipenser schrenckii)精子冷冻保存效果的影响及其冷冻损伤机理,比较了添加不同浓度海藻糖和蔗糖作为冷冻保护剂的精子稀释液处理后施氏鲟精子冷冻复苏的活力、快速运动时间和寿命。结果表明,添加60 mmol·L-1海藻糖1.0 mmol·L-1氯化钾的稀释液处理后,精子冻后快速运动时间和寿命与鲜精相比无显著差异(P>0.05);且活力较其他处理组有所提高,达到(26.67±3.32)%,但仍显著低于鲜精(P<0.05)。利用透射电镜和扫描电镜对施氏鲟精子超低温冷冻保存前后的超微结构进行观察,并对其能量代谢酶和抗氧化酶活进行测定和比较,结果表明,低温冻存造成施氏鲟精子的膜系统和细胞器(主要为线粒体和轴丝)损伤;能量代谢酶[总ATP酶、肌酸激酶(CK)、琥珀酸脱氢酶(SDH)、乳酸脱氢酶(LDH)]和抗氧化酶[过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)]活性均显著下降(P<0.05),而抗氧化酶谷胱甘肽还原酶(GR)活性显著上升(P<0.05),表明... 相似文献
3.
人工养殖西伯利亚鲟精子超低温冷冻保存研究 总被引：7，自引：6，他引：7
研究了人工养殖西伯利亚鲟精子的生物学特征及超低温冷冻保存方法。西伯利亚鲟的产精量为113.67±39.86 ml,精子密度为(6.49±3.10)×108/ml,精子活力为(85.4±9.5)%,精子寿命为353±23 s。精子密度与精子快速运动时间、精子寿命之间均存在线性相关,用方程分别表示为:y=1.0384x+1.5089(R2=0.7325);y=2.9069x+74.289(R2=0.6967)。结果表明精子密度可作为一项精子质量评价的标准。通过比较西伯利亚鲟精子在不同稀释液、不同抗冻剂和抗冻剂浓度、降温速率、解冻温度下的保存效果,结果表明:配方2作为稀释液,18%甲醇作为抗冻剂,二步法超低温(-196℃)冷冻保存精子,40℃水浴解冻取得最好的冻后活力,解冻后活力为(51.8±5.8)%。西伯利亚鲟授精的最佳精卵比为106∶1。在此精卵比下用冻精授精分别得到了(72.3±3)%的受精率和(52.9±4.1)%的孵化率,其中受精率与鲜精没有显著性差异,孵化率与鲜精有显著差异(P<0.05)。 相似文献
4.
以HBSS溶液为稀释液,DMSO为抗冻剂,0.25 mL麦细管为冻存管,两步降温法超低温冻存黄姑鱼精子,并用单细胞凝胶电泳(SCGE)技术检测了冻精的DNA损伤,荧光双染色流式细胞仪技术(FCM)检测了冻精的细胞膜性结构损伤。结果表明,DMSO质量分数在5%~20%时,冻精的活力与鲜精相比无显著差异;其中DMSO质量分数在10%时,冻精的激活率、运动时间及寿命分别为85.25%±3.95%、(3.23±0.27) min及(3.83±0.33) min。DMSO质量分数在25%、30%时,冻精的活力显著下降。SCGE检测显示,DMSO质量分数在5%~15%时、冻精的DNA损伤与鲜精相比差异不显著,DMSO质量分数为20%、25%、30%时,冻精的DNA损伤明显加重,冻精的DNA损伤与抗冻剂DMSO的质量分数成正相关。FCM检测显示,DMSO质量分数在5%~20%时,冻精中线粒体及细胞膜结构保持完整性的精子比例与鲜精相比无显著差异,DMSO质量分数在25%、30%时,冻精中的线粒体及细胞膜结构保持完整性的精子比例明显下降。分析认为,较高质量分数的DMSO是引起冻精活力下降,DNA、线粒体及细胞膜结构损伤加重的主要原因。 相似文献
5.
研究了超低温冷冻保存(-196℃)对俄罗斯鲟(Acipenser gueldenstaedti)精浆和精子中总三磷酸腺苷酶(AT-Pase)、琥珀酸脱氢酶(SDH)、乳酸脱氢酶(LDH)、肌酸激酶(CK)等酶活性的影响,并分别测定了冷冻前后俄罗斯鲟精浆和精子中酶的活性。结果显示,经过超低温冷冻保存后,俄罗斯鲟精子活力下降,精子内各酶活性均显著降低,添加冷冻保护液组精子中总ATPase、SDH、LDH和CK的活性分别从(198.47±14.43)U/mL、(30.00±2.65)U/mL、(6 982.29±24.32)U/L和(1.94±0.05)U/mL下降至(110.19±2.32)U/mL、(16.33±2.08)U/mL、(5 122.93±195.07)U/L和(1.49±0.14)U/mL。未添加抗冻剂组则分别下降至(2.25±0.33)U/mL、(11.67±0.58)U/mL、(4 488.04±78.33)U/L和(1.16±0.02)U/mL;精浆中酶的活性均显著升高,添加冷冻保护液组精浆总ATPase、SDH、LDH和CK活性分别从(12.70±0.57)U/mL、(7.50±0.71)U/mL、(2017.26±116.81)U/L和(2.93±0.59)U/mL升高至(92.49±5.18)U/mL、(13.33±0.58)U/mL、(3 688.97±172.67)U/L和(4.39±0.24)U/mL,未添加抗冻剂组则分别上升至(200.27±12.97)U/mL、(24.67±3.06)U/mL、(6 124.40±329.14)U/L和(5.20±0.16)U/mL。结果表明,超低温冷冻对俄罗斯鲟精浆和精子中酶活性及精子活力均有较大影响。 相似文献
6.
对瓦氏黄颡鱼(Pelteobagrus vachelli)精子在不同盐度和pH下的精子活力进行观察,同时研究了精子在4种不同稀释液与2种不同浓度抗冻剂组成的保存液中的超低温冷冻保存,并开展了冻精的授精实验。结果表明,瓦氏黄颡鱼精子浓度为(2.035±0.179)×1012cell·mL-1,在盐度为5.8、pH为7.17时,精子的活力都高达95%。以A液作为稀释液、10%甲醇作为抗冻剂时,冷冻保存精子效果最好,解冻后精子活力为(81.7±0.9)%。用解冻后的精子进行人工授精,获得的受精率为(88.4±2.1)%,孵化率为(74.0±0.8)%;而鲜精受精率为(91.0±0.8)%,孵化率(82±1.6)%,冻精与鲜精均无显著性差异。人工授精实验证明了解冻后的精子能正常用于该鱼的人工繁殖。 相似文献
7.
超低温冷冻对斑尾刺虾虎鱼卵中酶活性的影响 总被引：1，自引：0，他引：1
研究了超低温冷冻保存(-196℃)对斑尾刺虾虎鱼成熟卵中总ATP酶、肌酸激酶(CK)、琥珀酸脱氢酶(SDH)、乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-PX)和丙二醛(MDA)等酶活性的影响。结果表明,经过冷冻保存后,总ATP酶、CK、SDH和LDH的活性显著下降(P<0.05),同时可见添加抗冻剂组比未添加抗冻剂组下降幅度更大(P<0.05);SOD、CAT和GSH-PX的活性也显著下降(P<0.05),未添加抗冻剂组比添加抗冻剂组下降更明显(P<0.05);MDA的活性显著升高,且未添加抗冻剂组显著高于添加抗冻剂组(P<0.05)。超低温冷冻和是否添加抗冻剂都对斑尾刺虾虎鱼卵中酶活性有显著性影响。 相似文献
8.
9.
为建立圆口铜鱼(Coreius guichenoti)精子超低温冷冻保存方法,采用计算机辅助精子分析系统(CASA)评价了4种稀释液(D15、D20、L1、D1), 3种抗冻保护剂[二甲基甲酰胺(DMF)、二甲基亚砜(DMSO)、甲醇(METH)]在3种体积浓度(7.5%、10%、12.5%, V/V)下对圆口铜鱼精液的冷冻保存效果,并比较了150 mOsm/kg NaCl与超纯水作为激活剂对精子活力的影响。结果表明,D1组稀释液的精子活率(MOT)最高,为(74.64±13.17)%,与鲜精无显著差异(P0.05);以10%甲醇作为抗冻保护剂的圆口铜鱼精子经超纯水激活后,测得MOT最高,达到(78.11±14.74)%,与鲜精无显著差异(P0.05),精子运动速度达最大,平均曲线运动速度(VCL)、平均直线运动速度(VSL)、VAP(平均路径运动速度)分别达到(50.28±12.46)μm/s、(35.06±10.82)μm/s、(39.44±12.46)μm/s,精子快速运动时间和寿命分别为(8.67±1.15) s、(33.33±5.00) s,显著低于鲜精(P0.05); 7.5%甲醇组和7.5%二甲基甲酰胺组的MOT次之,分别为(77.71±17.74)%、(76.42±12.49)%,抗冻剂二甲基亚砜组的MOT显著低于甲醇组、二甲基甲酰胺组(P0.05)。12.5%的3种抗冻保护剂中,几乎无精子存活;在不同种类不同浓度的抗冻剂保护下,10%甲醇组,相较于使用超纯水激活,用150 mOsm/kg NaCl激活后的精子活率和寿命更高,说明Na~+有延长冻精寿命,提高精子活率的作用。研究表明, D1+10%METH (7.8 g/L NaCl+0.5 g/L KCl+15 g/L葡萄糖+10%METH)可用于圆口铜鱼精液超低温冷冻保存,为圆口铜鱼的繁育工作和种质资源保护奠定了基础。 相似文献
10.
《海洋渔业》2017,(6)
选取6 ind健康的雄性俄罗斯鲟(Acipenser gueldenstaedti),经人工催产后获得成熟的精子,运用扫描电镜和透射电镜,观察了超低温冷冻前后精子的形态结构。结果显示,经过超低温冷冻后精子形态结构发生了较大变化,其中冻精顶体长度、头中部宽度及中段宽度与鲜精相比显著增加(P0.05);中段长度及后外侧延伸物与鲜精相比显著减少(P0.05)。冻后有38.6%的精子在形态结构上受到不同程度的损伤。精子的结构损伤主要表现在精子顶体不明显或与核发生糅合,后外侧延伸物消失,甚至顶体脱落;精子核膜皱褶,泡状化,膜与核之间空隙加大,有的部分甚至出现膜断裂,核内内切沟模糊;精子中段发生膨大和变形,线粒体外膜破损,线粒体条状嵴结构弥散,线粒体脱落;鞭毛外鳍褶与鞭毛脱离,鞭毛与中段脱离,少数鞭毛在中段断开。结果表明,超低温冷冻对精子的损伤主要集中在膜系统,中心粒等微管系统基本完好。 相似文献
11.
Cryopreservation of Arctic charr Salvelinus alpinus (L.) semen was investigated using three diluents, three cryoprotectants [10% dimethyl sulphoxide (DMSO), 10% dimethylacetamide (DMA) or 20% glycerol] and three sizes of straw. The three diluents and three cryoprotectants were combined, resulting in nine extenders. One part semen was added to three parts extender, and motility was evaluated to assess the toxicity of six of the extenders. Semen in nine extenders was frozen in 0.5‐mL straws using liquid nitrogen vapour. Semen extended in 0.3 m glucose and each of the cryoprotectants was also frozen in 0.5‐mL, 1.7‐mL (flat) or 2.5‐mL straws. The freezing rate in each size of straw was measured. Fertility trials were conducted to determine the post‐thaw viability of the frozen semen. The motility of activated spermatozoa was higher in the DMA and DMSO extenders than in the glycerol extender. For the trial using 0.5‐mL straws, post‐thaw fertility results were higher for all extenders containing DMSO, or 0.3 m glucose and DMA, than for all other combinations of diluent and cryoprotectant. For the straw size comparison, the highest fertility was obtained for the 1.7‐mL straw using either DMSO or DMA and for the 2.5‐mL straw using DMSO. For all cryopreservation trials, fertility was low for extenders containing glycerol. 相似文献
12.
为筛选出更有效的虹鳟(Oncorhynchus mykiss)精液冷冻保存液,在以10％ 甲醇和0.3 M葡萄糖作为主要成分的基础上,分别或同时添加0.9 g/L氯化钾和10％ 蛋黄形成四种冷冻保存液,用于虹鳟精液冷冻保存试验,分别形成4个处理组:Ⅰ#(不添加氯化钾和蛋黄)、Ⅱ#(添加氯化钾)、Ⅲ#(添加蛋黄)和Ⅳ#(... 相似文献
13.
Cryopreservation of sperm in marine fish 总被引：10，自引：0，他引：10
Since the first work of Blaxter in 1953, fish sperm cryopreservation has been attempted on about 30 marine species. The present paper reviews the techniques used and the results published in these species. Particular attention is paid to the handling procedure of sperm before freezing, the problems of semen ageing and semen contamination with urine. The quality of frozen–thawed semen was evaluated using previously standardized biotests, such as a two‐step motility activation technique adapted for the different species and fertilization assays using a discriminating insemination technique. Most extenders used in marine fish are saline or sugar solutions. From the investigated cryoprotectants, dimethyl sulphoxide (DMSO) generally leads to the best results. Cooling rates range from 8 °C to 99 °C min?1; the thawing rate is generally high. Compared with freshwater species, a high percentage of spermatozoa survives cryopreservation. Therefore, and because of the simplicity of the techniques, the cryopreservation of marine fish sperm is suited for application in aquaculture. 相似文献
14.
The effects of extender composition and freezing rate on motility and fertility of frozen‐thawed Arctic char, Salvelinus alpinus, spermatozoa were investigated. Three freezing rates, two semen diluents and three cryoprotectants were tested. Semen frozen in 0.3 mol L?1 glucose diluent with 10% methanol as a cryoprotectant or in a diluent described by Lahnsteiner with 10%N,N‐dimethylacetamide (DMA) resulted in the highest sperm motility. Fertility was the highest for semen frozen in a glucose–methanol extender but was not significantly different than that for semen frozen in Lahnsteiner's diluent with 10% DMA. Dimethyl sulphoxide (DMSO) at 10% was a relatively ineffective cryoprotectant with either semen diluent. Semen frozen at 6 cm above the surface of liquid nitrogen resulted in a higher post‐thaw sperm motility and fertility than semen frozen at 5 cm. The addition of 7% fresh egg yolk to glucose diluent containing methanol or DMSO did not improve the fertility of frozen‐thawed spermatozoa. However, the addition of 7% fresh egg yolk to glucose–DMA extender significantly improved the fertilization percentages of frozen‐thawed spermatozoa. In conclusion, dilution of semen 1:3 in 0.3 mol L?1 glucose with 10% methanol and freezing 6 cm above the surface of liquid nitrogen (freezing rate of 40±8°C min?1, mean±SD from ?5 to ?55°C) is a promising protocol for cryopreservation of Arctic char semen. 相似文献
15.
A.N. Maria A.T.M. Viveiros R.T.F. Freitas A.V. Oliveira 《Aquaculture (Amsterdam, Netherlands)》2006,260(1-4):298-306
The aim of this study was to develop a protocol for semen storage of piracanjuba (Brycon orbignyanus) by both cool storage at 4 °C and cryopreservation at − 196 °C. Semen was diluted in some fish semen extenders (Exp. 1) or in extenders combined with the antibiotic gentamycin sulfate (Exp. 2) and stored at 4 °C. Sperm motility was estimated every 24 h. Then, the effects of egg yolk (0 and 5%), cryoprotectants (dimethyl sulphoxide — DMSO, methanol, and methylglycol) and extenders (NaCl 154 mM, BTS™ Minitub and M III™ Minitub) on semen cryopreservation were evaluated (Exp. 3). Semen was added to each of eighteen cryosolutions (2 yolk concentrations × 3 cryoprotectants × 3 extenders), aspirated into 0.5-mL straws, frozen in nitrogen vapor (Taylor-Wharton, CP 300, “dry shipper”) and stored at − 196 °C. Sperm motility was evaluated after thawing at 60 °C-water bath for 8 s. The three cryosolutions that produced the highest post-thaw sperm motility were used again to freeze semen. Post-thaw semen quality was then evaluated under three tests: sperm motility, the percentage of live spermatozoa and hatching rate (Exp. 4). Piracanjuba semen diluted (1:10 total volume) in NaCl 200 mM or in Saad solution (NaCl 200 mM, Tris 30 mM) maintained motility above 35% for as long as 7 days, at 4 °C. Motility of only 7% was observed on undiluted semen after 3 days at 4 °C. There was neither beneficial nor detrimental effect of gentamycin on sperm motility at 250 μg/mL. Egg yolk addition to the cryosolution was beneficial in samples cryopreserved in NaCl 154 mM and in M III™, but detrimental for samples cryopreserved in BTS™. Methylglycol was the most effective cryoprotector compared to DMSO and methanol. Motility and percentage of live spermatozoa were similar among semen cryopreserved in NaCl–yolk, M III™–yolk and BTS™, all containing 10% methylglycol, but lower than fresh control. Hatching rates of eggs fertilized with sperm cryopreserved in NaCl–yolk or BTS™ were higher than for eggs fertilized with sperm cryopreserved in M III™–yolk, but lower than control fertilizations. The semen cryopreservation protocols developed here will be used to set up a gene bank for endangered piracanjuba populations. 相似文献
16.
Abstract. The effects of varying temperature, salinity and pH on the activation and subsequent motility of sperm of the yellowfin bream, Acanthopagrus australis (Günther), were assessed using a linear scale based on the overall activity of the sperm over time. Motility half-life was calculated using log transformation.
Conditions reflecting the natural habitat of the fish, oceanic salinity (1200 mOsM) and slight alkalinity (pH 8-8-5), were shown to produce both maximum activation and subsequent motility duration. The half-life of activated sperm was shown to be greater at 4°C than at 20-23°C (5·13 min:22·8 min). Storage of freshly stripped semen was shown to be most successful at 4°C with a half-life of 98·9 min.
The cryopreservation of semen was tested using the cryoprotectants glycerol and dimethyl sulphoxide at concentrations ranging from 0·5M to 2·0M, and pre-freezing equilibration times of 5 and 15 min. Glycerol at 20 M was shown to give significantly superior results. There was no significant difference between sperm activation or sperm half-life for fresh-stripped semen and frozen semen, using glycerol at 2·0M as the cryoprotectant. 相似文献
Conditions reflecting the natural habitat of the fish, oceanic salinity (1200 mOsM) and slight alkalinity (pH 8-8-5), were shown to produce both maximum activation and subsequent motility duration. The half-life of activated sperm was shown to be greater at 4°C than at 20-23°C (5·13 min:22·8 min). Storage of freshly stripped semen was shown to be most successful at 4°C with a half-life of 98·9 min.
The cryopreservation of semen was tested using the cryoprotectants glycerol and dimethyl sulphoxide at concentrations ranging from 0·5M to 2·0M, and pre-freezing equilibration times of 5 and 15 min. Glycerol at 20 M was shown to give significantly superior results. There was no significant difference between sperm activation or sperm half-life for fresh-stripped semen and frozen semen, using glycerol at 2·0M as the cryoprotectant. 相似文献
17.
达氏鲟精巢细胞消化分离和超低温冷冻保存 总被引：1，自引：0，他引：1
通过研究两种酶对精巢细胞的消化效果,探究抗冻剂、降温程序、糖类和卵磷脂对达氏鲟(Acipenser dabryanus)精巢细胞冻存效果的影响,获得消化冻存后高存活率的细胞。实验使用0.25%胰蛋白酶和2 mg/m L胶原酶H+500 U/m L中性酶II的组合酶对达氏鲟精巢细胞进行消化,获得不同消化时间内的活细胞数量。另外,还研究了冷冻稀释液中分别添加10%二甲基亚砜(DMSO)、乙二醇(EG)、甲醇(MET)作为抗冻剂,采用-1℃/min慢速降温以及直接投入液氮中的快速降温方法冻存,冷冻稀释液采用D-海藻糖或同浓度D-蔗糖,以及添加5%、8%、11%卵磷脂对冻存效果的影响。结果显示:两种消化酶在同一时间消化所得的活细胞数和活细胞率无显著差异,并且都在3 h获得最多活细胞。慢速降温的冻存效果极显著地好于快速降温(P=0.01)。不同抗冻剂的保存效果差异显著,复苏后细胞相对存活率EG(51.70%±5.24%)MET(45.09%±3.15%)DMSO(40.18%±3.90%)。不同糖对达氏鲟精巢细胞冻存效果无显著影响;不同浓度的卵磷脂冻存效果有极显著差异。含8%卵磷脂的冻存液对细胞的冻存效果最好,细胞存活率可达(93.55±2.56)%,培养10 d后细胞数目为0 d时的3.19倍。 相似文献
18.
R. Paul Lang Kenneth L. Riley John E. Chandler Terrence R. Tiersch 《Journal of the World Aquaculture Society》2003,34(1):66-75
The commercial‐scale production of fish by use of artificial (induced) spawning would require reliable, large‐volume sources of sperm. Cryopreservation can be used to preserve and store sperm within commercial and research germplasm repositories, but is limited in its application to aquaculture. Straw volume and cooling chamber size restrict the quantity of sperm that can be frozen, and straws must be filled by hand. In contrast, the dairy industry has refined methods for freezing of bull sperm, including automation of straw filling and the use of large cooling chambers. These methods could be used for commercial‐scale cryopreservation of fish sperm, although application would require testing. To supply sperm in large volumes, bags originally developed for swine semen could be cooled using dairy protocols and used as a container for fish sperm. The current study documented the use of commercial‐scale dairy cryopreservation techniques for the production of hybrids of channel catfish Ictalurus punctatus (female) by blue catfish Ictalurus furcarus. Four cryoprotectants (methanol, dimethyl sulfoxide, dimethyl acetamide, and glycerol) were initially evaluated for use with blue catfish sperm. During May 2000 and March to April 2001, suspensions of blue catfish sperm were cryopreserved with 10% methanol in 0.5‐mL French straws and in commercial swine semen bags (Cochette* bags, IMV International. Minneapolis, Minnesota, USA). Cryopreservation took place at a dairy breeding cooperative, using technology employed for bull semen. Sperm motility before freezing was 26 ± 18% during Year 1 (2000) and 62 ± 30% during 2001. Sperm were thawed at 40 C and used to fertilize the eggs of channel catfish (yielding hybrids). Motility after thawing for sperm frozen in 0.5‐mL straws was 11 ± 10% during 2000 and 50 ± 24% during 2001. Motility after thawing was 41 ± 17% for sperm frozen in swine semen bags in 5‐mL aliquots and 43 ± 10% for sperm frozen in 10‐mL aliquots. Neurulation of eggs fertilized with thawed sperm from straws was 83 ± 13% during 2000 and 54 ± 27% during 2001. Neurulation was 57 ± 24% using sperm frozen in swine semen bags in 5‐mL aliquots and 55 ± 10% using sperm frozen in 10‐mL aliquots. There was no correlation between sperm motility before freezing (in 0.5‐mL straws) and after thawing during 2000 (r= 0.52) or during 2001 (r= 0.49). In addition, there was no correlation between initial motility and neurulation of channel catfish eggs fertilized using thawed sperm during 2000 (r= 0.14) or during 2001 (r= 0.29). Sperm of blue catfish can thus be cryopreserved at a commercial scale using dairy protocols and can be made available for the production of hybrid catfish when viable eggs are available. 相似文献
19.
Sperm cryopreservation has led to transcendental changes in the reproductive biotechnology of both mammals and fish, and is a basic tool for animal improvement. However, these protocols generate damage to cell structure and physiology, altering sperm function as a result of cryoinjuries during freezing and thawing. This review is a compilation of the techniques developed and standardised for assessing sperm function in cryopreserved fish semen. Recent studies have analysed sperm function objectively, applying cellular and molecular techniques to characterise cryodamage. The Computer Assisted Sperm Analysis system has facilitated the assessment of motility, while electron microscopy (SEM and TEM) and cryo‐microscopy have made it possible to study sperm morphology and ultra‐structure. The effects of cryodamage on nuclear DNA have also been analysed using various methods, including the comet Fluorescence in situ Hybridization test, TUNEL, Sperm Chromatin Structure Assay, specific DNA sequences using RT‐PCR and specific genes by qPCR. The latter technique is used to study the mitochondrial genome (mtDNA), together with some candidate genes which are associated with bioenergy activity and sperm motility. Other parameters assessed are mitochondrial membrane potential and ATP content using high performance liquid chromatography, nuclear magnetic resonance spectrometry and cell respiration. All this information makes it possible to establish study and assessment criteria for cryopreserved fish spermatozoa. This work focuses on the use of technologies to study of quality of fish spermatozoa during cryopreservation. 相似文献
20.
采用0、100、200、400、800μmol/L的硫酸铜和0、200、400、800、1600μmol/L的氯化锌分别处理泥鳅鳍细胞系,培养24 h后,用单细胞凝胶电泳检测其DNA损伤情况,同时对金属硫蛋白基因的表达进行研究。试验结果表明,随着两种重金属浓度的增加,泥鳅鳍细胞的DNA损伤程度呈现明显的剂量依赖性。当硫酸铜浓度≥200μmol/L、氯化锌浓度≥400μmol/L时,细胞的拖尾率、彗尾DNA比例、彗星尾长、彗星尾距和Olive尾矩均显著增大(P<0.05)。泥鳅细胞金属硫蛋白基因的表达量随着硫酸铜浓度的升高呈现出短暂稳定后显著下降的趋势,而随着氯化锌浓度的增大呈现出显著上升再显著下降的趋势,表明氯化锌浓度≤400μmol/L时可显著诱导泥鳅鳍细胞系金属硫蛋白的转录表达。综合来看,与锌相比,泥鳅鳍细胞对铜的解毒能力和耐受力更差。因此,铜污染对泥鳅细胞的遗传毒性更加明显。本研究将为探讨重金属对生物的毒性效应及生物学监测提供理论依据。 相似文献