Froman D P, Engel H N
Department of Poultry Science, Oregon State University, Corvallis 97331.
Biol Reprod. 1989 Mar;40(3):615-21. doi: 10.1095/biolreprod40.3.615.
The hypothesis that sialic acid has a role in spermatozoal sequestration within the hen's oviduct was tested by treating spermatozoa with Clostridium perfringens neuraminidase. Spermatozoal content of sialic acid ranged from 94 to 135 micrograms per 10(9) spermatozoa (n = 12 roosters). Spermatozoa contained 80% of total seminal sialic acid (coefficient of variation = 4.6%). Spermatozoal sialic acid content was reduced by 18% when 10(9) spermatozoa were incubated at pH 6.5 with 10 IU neuraminidase activity (Type V, Sigma Chemical Co.). Such treatment had no effect on spermatozoal viability as evidenced by ethidium bromide uptake. However, treatment of spermatozoa with neuraminidase prior to intravaginal insemination reduced fertility by 24 percentage units (p less than 0.001). In contrast, when similarly treated spermatozoa were deposited in the magnum via laparatomy, fertility was not affected (p greater than 0.05). The preceding work was done with neuraminidase prepared by salt fractionation (Type V, Sigma Chemical Co.). Type V neuraminidase was absorbed to diethylaminoethyl-Sephacel and then eluted with a stepwise KCl gradient. Treatment of spermatozoa with this preparation of neuraminidase (10 IU/10(9) spermatozoa) prior to intravaginal insemination reduced fertility by 19 percentage units (p less than 0.001). Decreased fertility could not be attributed to contamination of neuraminidase preparation with proteolytic activity. We conclude that spermatozoal sialic acid has a role in spermatozoal sequestration within the hen's utero-vaginal glands.
通过用产气荚膜梭菌神经氨酸酶处理精子,对唾液酸在母鸡输卵管内精子滞留中起作用这一假说进行了验证。每10⁹个精子中唾液酸的含量在94至135微克之间(n = 12只公鸡)。精子含有精液中总唾液酸的80%(变异系数 = 4.6%)。当10⁹个精子在pH 6.5下与10 IU神经氨酸酶活性(V型,西格玛化学公司)一起孵育时,精子唾液酸含量降低了18%。如溴化乙锭摄取所示,这种处理对精子活力没有影响。然而,在阴道内输精前用神经氨酸酶处理精子会使受精率降低24个百分点(p < 0.001)。相比之下,当将经过类似处理的精子通过剖腹术放入输卵管壶腹部时,受精率不受影响(p > 0.05)。之前的工作是用通过盐分级分离制备的神经氨酸酶(V型,西格玛化学公司)完成的。V型神经氨酸酶吸附到二乙氨基乙基 - 葡聚糖凝胶上,然后用逐步的氯化钾梯度洗脱。在阴道内输精前用这种神经氨酸酶制剂(10 IU/10⁹个精子)处理精子会使受精率降低19个百分点(p < 0.001)。受精率降低不能归因于神经氨酸酶制剂被蛋白水解活性污染。我们得出结论,精子唾液酸在母鸡子宫 - 阴道腺内精子滞留中起作用。