Sensitivity of domestic cat (Felis catus) sperm from normospermic versus teratospermic donors to cold-induced acrosomal damage.
Sensitivity of domestic cat (Felis catus) sperm from normospermic versus teratospermic donors to cold-induced acrosomal damage.
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正常精子与畸形精子供体的家猫（Felis catus）精子对寒冷引起的顶体损伤的敏感性。
DOI：
10.1095/biolreprod61.1.135
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发表时间：
1999
影响因子：
3.6
通讯作者：
Howard,J
中科院分区：
文献类型：
--
作者：
Pukazhenthi,B;Pelican,K;Wildt,D;Howard,J
猫精子在冷冻保护剂中冻融会导致广泛的膜损伤。为了确定单独冷却是否会影响精子结构和活力，我们比较了冷却速率对正常精子（N;每次射精> 60%正常精子）和畸形精子（T;每次射精< 40%正常精子）家猫精子的影响。将电射精分成未处理的或洗涤的（Ham's F-10 + 5%胎牛血清）等分试样，将后者重悬于Ham's F-10培养基或无甘油过滤的Platz稀释剂变体（20%蛋黄，11%乳糖）中。将等分试样1）保持在25°C（不冷却;对照），2）在商业冰箱中冷却至5°C持续30分钟（快速冷却; 0.14 °C/min），3）在0°C下置于冰泥中持续10分钟（超快速冷却; 0.14 °C/min），或4）在可编程醇浴中以0.5°C/min冷却至0°C（缓慢冷却）;并且每4°C取出等分试样。然后将所有样品加热至25°C，并使用荧光素缀合的花生凝集素染色剂评价精子活力百分比和完整顶体的比例。在两个猫种群中，暴露于低温和升温至25°C后，精子活力百分比立即保持不受影响（p> 0.05）。然而，与对照值（N，81.5%; T，77.5%）相比，快速冷却（104 °C/min）至5°C（N，65.6%; T，27.5%）或超快速冷却（1014 °C/min）至0°C（N，62.1%; T，23.0%）后，具有完整顶体的精子比例下降（p< 0.05）。冷却精子的透射电子显微镜显示顶体膜广泛损伤。相反，缓慢冷却（0.5°C/min）至5°C可维持（p> 0.05）高比例的顶体完整的精子（N，75.5%; T，68.3%），通过继续冷却至0°C，这在猫种群之间也保持相似（p> 0.05）（N，64.7%; T，56.8%）。结果表明：1）快速冷却家猫精子可诱导显著的顶体损伤，但不影响精子活力; 2）畸形精子症雄性精子比正常精子症雄性精子更容易受到冷诱导的顶体损伤; 3）降低初始冷却速率可显著降低精子结构损伤。
Freeze-thawing cat sperm in cryoprotectant results in extensive membrane damage. To determine whether cooling alone influences sperm structure and viability, we compared the effect of cooling rate on sperm from normospermic (N; > 60% normal sperm per ejaculate) and teratospermic (T; < 40% normal sperm per ejaculate) domestic cats. Electroejaculates were divided into raw or washed (Ham's F-10 + 5% fetal calf serum) aliquots, with the latter resuspended in Ham's F-10 medium or Platz Diluent Variant Filtered without glycerol (20% egg yolk, 11% lactose). Aliquots were 1) maintained at 25°C (no cooling; control), 2) cooled to 5°C in a commercial refrigerator for 30 min (rapid cooling; ∼4°C/min), 3) placed in an ice slush at 0°C for 10 min (ultrarapid cooling; ∼14°C/min), or 4) cooled to 0°C at 0.5°C/min in a programmable alcohol bath (slow cooling); and aliquots were removed every 4°C. All samples then were warmed to 25°C and evaluated for percentage sperm motility and the proportion of intact acrosomes using a fluorescein-conjugated peanut agglutinin stain. In both cat populations, sperm percentage motility remained unaffected (p> 0.05) immediately after exposure to low temperatures and after warming to 25°C. However, the proportion of spermatozoa with intact acrosomes declined (p< 0.05) after rapid cooling (∼4°C/min) to 5°C (N, 65.6%; T, 27.5%) or ultrarapid cooling (∼14°C/min) to 0°C (N, 62.1%; T, 23.0%) in comparison to the control value (N, 81.5%; T, 77.5%). Transmission electron microscopy of cooled sperm revealed extensive damage to acrosomal membranes. In contrast, slow cooling (0.5°C/min) to 5°C maintained (p> 0.05) a high proportion of spermatozoa with intact acrosomes (N, 75.5%; T, 68.3%), which also remained similar (p> 0.05) between cat populations (N, 64.7%; T, 56.8%) through continued cooling to 0°C. Results demonstrate that 1) rapid cooling of domestic cat sperm induces significant acrosomal damage without altering sperm motility, 2) spermatozoa from teratospermic males are more susceptible to cold-induced acrosomal damage than normospermic counterparts, and 3) reducing the rate of initial cooling markedly decreases sperm structural damage.