ZONA-PELLUCIDA FILTRATION OF STRUCTURALLY ABNORMAL SPERMATOZOA AND REDUCED FERTILIZATION IN TERATOSPERMIC CATS
ZONA-PELLUCIDA FILTRATION OF STRUCTURALLY ABNORMAL SPERMATOZOA AND REDUCED FERTILIZATION IN TERATOSPERMIC CATS
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DOI：
10.1095/biolreprod49.1.131
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发表时间：
1993-07-01
影响因子：
3.6
通讯作者：
WILDT, DE
中科院分区：
文献类型：
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作者：
HOWARD, JG;DONOGHUE, AM;WILDT, DE
用家猫的正常精子(60%结构正常精子/射精)和畸形精子(40%结构正常精子/射精)精液检测透明带(ZP)穿透、体外受精、胚胎发育和受精精子的形态。此外，还比较了上游处理与简单精子洗涤对精子-卵母细胞相互作用的影响。对正常精子和畸精症精液的精子活力和形态进行了评估。将精子预培养1h，然后与体内成熟的猫卵泡卵母细胞(n=401)共孵育20h，与ZP完整的盐保存卵母细胞(n=202)共孵育6h，评估体内成熟卵母细胞的卵裂率和胚胎发育阶段。对盐保存卵母细胞的ZP穿透率(内ZP内或穿过内ZP的卵母细胞比例)、内ZP穿透精子的平均(+/-SEM)数以及所有结合和穿透精子的形态进行了评估。正常精子症患者生精液中多形精子的发生率平均为29%，而畸形症患者为67%，但所有精液中的精子活动率都很高(60%)。上游处理增加了(p<；0.05)正常精子回收/畸形精子人工授精的数量(66.5+/-2.3%)，而简单洗涤后的回收(28.6+/-2.2%)。男性畸形症患者上游漂流后精子活动率(90.0+/-1.3%)也高于洗精组(64.2+/-3.7%)(p<；0.05)。来自正常精子的精子的体外卵裂率(86.3%)高于畸形精子(50.3%)的体外卵裂率(p<；0.05)，但胚胎发育率与桑拿期/囊胚期相似(p>；0.05)。正常精子射精液对盐保存卵母细胞的ZP穿透率(73.7%)高于畸形精子射精液(24%)(p<；0.01)。正常精子组ZP穿透精子/卵母细胞数是畸形组的5倍(p<；0.05)。结构正常的结合精子和ZP穿透精子的比例在不同的猫种群之间是相似的(p>；0.05)，无论授精的形态如何。结构异常的畸形精子能与ZP结合(29%)，并能初步穿透到外ZP(1.7%)。然而，只有3%的内ZP精子具有多形性，卵黄周间隙内的每个精子形态正常。尽管上游处理增加了精子活力和结构正常的精子数量，但在卵裂或ZP穿透方面没有观察到差异(p>；0.05)。这些结果表明，ZP是过滤结构异常精子的有效过滤器，但即使是来自畸形精子症猫的正常精子，受精能力也似乎受到了损害。尽管畸形精子症影响了ZP的穿透和受精，但通过畸形精子射精液产生的少数胚胎在体外发育正常。
Zona pellucida (ZP) penetration, in vitro fertilization, embryo development, and the morphology of fertilizing sperm were examined through use of normospermic (> 60% structurally normal sperm/ejaculate) versus teratospermic (< 40% structurally normal sperm/ejaculate) ejaculates from domestic cats. In addition, the effect of swim-up processing on sperm-oocyte interaction was compared with that of simple sperm washing. Normospermic and teratospermic ejaculates were evaluated for sperm motility and morphology. Sperm were preincubated for 1 h, then coincubated with in vivo-matured follicular cat oocytes (n = 401 ) for 20 h and with ZP-intact, salt-stored oocytes (n = 202) for 6 h. In vivo-matured oocytes were assessed for percent cleavage and stage of embryo development over time. Salt-stored oocytes were assessed for percent ZP penetration (proportion of oocytes containing sperm within or through the inner ZP), mean (+/- SEM) number of inner ZP-penetrated sperm, and the morphology of all bound and penetrated sperm. The incidence of pleiomorphic sperm in raw ejaculates averaged 29% in normospermic versus 67% in teratospermic males, but all ejaculates contained high sperm motility ratings (> 60%). Swim-up processing increased (p < 0.05) the number of normal sperm recovered/teratospermic inseminate (66.5 +/- 2.3%) compared to recovery after simple washing (28.6 +/- 2.2% ). Percent sperm motility also increased (p < 0.05) in teratospermic males after swim-up (90.0 +/- 1.3%) as compared to sperm washing (64.2 +/- 3.7%). Cleavage rate in vitro was higher (p < 0.05) using sperm from normospermic (86.3% ) compared to teratospermic (50.3% ) males, but rates of embryo development to the morula/blastocyst stage were similar (p > 0.05). ZP penetration of salt-stored oocytes by normospermic ejaculates (73.7% ) was superior (p < 0.01) to that of teratospermic ejaculates (24. 1% ); the number of ZP-penetrated sperm/oocyte was 5-fold higher (p < 0.05) in the normospermic than in the teratospermic group. The proportion of structurally normal bound and ZP-penetrated sperm was similar (p > 0.05) between cat populations regardless of the morphologic forms in the inseminate. Structurally abnormal sperm from teratospermic males were capable of ZP binding (29% ) and initial penetration into the outer ZP ( 1 7% ). However, only 3% of the inner ZP sperm were pleiomorphic, and every sperm within the perivitelline space was morphologically normal. Although swim-up processing increased sperm motility and the number of structurally normal sperm in the teratospermic inseminates, no difference (p > 0.05) was observed in cleavage or ZP penetration. These results indicate that the ZP is an efficient filter for structurally abnormal sperm, but even normal sperm from teratospermic cats appear impaired in fertilizing ability. Although teratospermia compromises ZP penetration and fertilization, the few embryos produced via teratospermic ejaculates develop normally in vitro.