Cryopreservation of Atlantic salmon Salmo salar sperm: effects on sperm physiology
Cryopreservation of Atlantic salmon Salmo salar sperm: effects on sperm physiology
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DOI：
10.1111/jfb.13052
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发表时间：
2016-09-01
影响因子：
2
通讯作者：
Farias, J. G.
中科院分区：
文献类型：
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作者：
Figueroa, E.;Valdebenito, I.;Farias, J. G.
本研究的目的是确定冷冻对大西洋鲑鱼萨尔莫salar精子功能的影响。将精液冷冻在科特兰氏培养基+1.3M二甲基亚砜+0.3M葡萄糖+2%牛血清白蛋白（终浓度）中，以精液：冷冻保护剂= 1：3为处理组（T），新鲜精液为对照组（F）。将0.5ml精子悬液的吸管冷冻在4cm的N_2L中。将它们在温度调节浴（40 ° C）中解冻。解冻后，用流式细胞术检测精子DNA片段化百分率（TUNEL法）、质膜完整性（SYBR-14/PI）和线粒体膜电位（MMit，JC-1），并在频闪光下用光学显微镜检测精子活力。在精子密度为15 × 10（7）个精子/卵母细胞（-1）的条件下，通过在10 ℃下孵育16小时后观察第一次卵裂来测试对照和处理精液的受精率。在冷冻保存的精液（T）中，平均值+/- s.d. DNA断裂48 ± 25%;质膜完整性752 ± 63%;线粒体膜电位517 ± 36%;运动性585 ± 53%;曲线速度（V-CL）612 ± 174 μ ms（-1）;平均路径速度（V-AP）501 ± 173 μ ms（-1）;直线流速（V-SL）591 ± 184 μ ms（-1），受精率816 ± 19%。质膜完整性、线粒体膜电位、运动性、受精率、V-CL、V-AP和V-SL与对照组相比差异显著（P
The objective of this study was to determine the effect of freezing on the function in Atlantic salmon Salmo salar spermatozoa. The semen was frozen in Cortland's medium+1.3M dimethyl sulphoxide+0.3M glucose+2% bovine serum albumin (final concentration) in a ratio of 1:3 (semen:cryoprotectant) as the treatment (T) and fresh semen as the control (F). Straws of 05ml of sperm suspension were frozen in 4cm of N2L. They were thawed in a thermoregulated bath (40 degrees C). After thawing, the percentage of spermatozoa with fragmented DNA [transferase dUTP (deoxyuridine triphosphate) nick-end labelling (TUNEL)], plasma membrane integrity (SYBR-14/PI) and mitochondrial membrane potential (MMit, JC-1) were evaluated by flow cytometry and motility was evaluated by optical microscope under stroboscopic light. The fertilization rates of the control and treatment semen were tested at a sperm density of 15x10(7) spermatozoa oocyte(-1), by observation of the first cleavages after 16h incubation at 10 degrees C. In the cryopreserved semen (T), the mean +/- s.d. DNA fragmentation was 48 +/- 25%; plasma membrane integrity 752 +/- 63%; mitochondrial membrane potential 517 +/- 36%; motility 585 +/- 53%; curved line velocity (V-CL) 612 +/- 174 mu ms(-1); average-path velocity (V-AP) 501 +/- 173 mu ms(-1); straight-line velocity (V-SL) 591 +/- 184 mu ms(-1); fertilization rate 816 +/- 19%. There were significant differences in the plasma membrane integrity, mitochondrial membrane potential, motility, fertilization rate, V-CL, V-AP and V-SL compared with the controls (P