Ellerman D A, Brantúa V S, Martínez S P, Cohen D J, Conesa D, Cuasnicú P S
Instituto de Biología y Medicina Experimental, Buenos Aires, Argentina.
Biol Reprod. 1998 Nov;59(5):1029-36. doi: 10.1095/biolreprod59.5.1029.
Rat epididymal protein DE associates with the sperm surface during maturation and participates in sperm-egg fusion. Immunization of male rats with DE raised specific antibodies and produced a significant reduction in the animals' fertility. The present study focused on determining the in vivo mechanism involved in fertility inhibition. Wistar males were injected with DE, and antibody levels and animal fertility were evaluated. Results revealed an association between the two parameters, since animals with absorbance values lower than 0.5 in ELISA presented high fertility rates (66%, 100%) while those with absorbance values higher than 0.5 exhibited the lowest fertility rates (0%, 33%). Histological studies showed no evidence of orchitis, epididymitis, or vasitis in DE-immunized animals. ELISA results revealed the presence of anti-DE antibodies in epididymal and vas deferential fluids. Indirect immunofluorescence and ELISA experiments indicated that these antibodies would not interfere with the synthesis or secretion of DE or with its association with the sperm surface. Finally, while epididymal sperm recovered from DE-immunized animals presented no changes in motility, viability, or ability to undergo capacitation and acrosome reaction, they exhibited a significant decrease in their ability to fuse with zona-free eggs, with no effect on their ability to bind to the oolemma. Together these results indicate that immunization of male rats with epididymal protein DE specifically interferes with the sperm fertilizing ability, supporting the use of epididymal proteins for contraceptive vaccine development.
大鼠附睾蛋白DE在精子成熟过程中与精子表面结合,并参与精卵融合。用DE免疫雄性大鼠可产生特异性抗体,并显著降低动物的生育能力。本研究重点在于确定生育抑制所涉及的体内机制。给Wistar雄性大鼠注射DE,并评估抗体水平和动物生育能力。结果显示这两个参数之间存在关联,因为在ELISA中吸光度值低于0.5的动物具有较高的生育率(66%,100%),而吸光度值高于0.5的动物生育率最低(0%,33%)。组织学研究表明,DE免疫的动物没有睾丸炎、附睾炎或输精管炎的迹象。ELISA结果显示附睾和输精管液中存在抗DE抗体。间接免疫荧光和ELISA实验表明,这些抗体不会干扰DE的合成或分泌,也不会干扰其与精子表面的结合。最后,虽然从DE免疫的动物中回收的附睾精子在活力、生存能力、获能和顶体反应能力方面没有变化,但它们与无透明带卵子融合的能力显著下降,而对其与卵膜结合的能力没有影响。这些结果共同表明,用附睾蛋白DE免疫雄性大鼠会特异性干扰精子的受精能力,支持将附睾蛋白用于避孕疫苗的开发。