Pimenta J, Dias F M V, Marques C C, Baptista M C, Vasques M I, Horta A E M, Barbas J P, Soares R, Mesquita P, Cabrita E, Fontes C M G A, Prates J A, Pereira R M
Unidade de Recursos Genéticos, Reprodução e Melhoramento Animal, Instituto Nacional dos Recursos Biológicos, Vale de Santarém, Portugal.
Reprod Domest Anim. 2012 Apr;47(2):196-202. doi: 10.1111/j.1439-0531.2011.01827.x. Epub 2011 Aug 2.
The function of prion-like protein Doppel was suggested to be related to male fertility. In this study, the importance of ovine Doppel polypeptide on spermatozoa capacitation and fertilization was evaluated. After refolding, recombinant Doppel (rDpl) was supplemented with different concentrations (40, 80 or 190 ng/ml) to ovine spermatozoa during the capacitation process. In experiment 1, post-thawed ovine spermatozoa were incubated with different concentrations of rDpl during 1 h for swim-up, and changes in sperm motility, concentration, vigour, viability and capacitation were monitored (10 replicates). In experiment 2, the fertilization ability of post-swim-up spermatozoa incubated as above was tested through heterologous fertilization of bovine in vitro matured oocytes (n = 423, three replicates). Regardless of dosage, rDpl improved (p ≤ 0.03) spermatozoa viability. Sperm individual motility and vigour were the highest (p ≤ 0.04) for the group receiving 190 ng/ml rDpl. Sperm supplemented with the highest doses of rDpl achieved higher (p = 0.02) fertilization rates (56.0 ± 3.0%) than control (39.1 ± 2.2%) and 40 ng/ml rDpl (39.8 ± 2.7%). Preliminary data suggest that Doppel protein may enhance in vitro spermatozoa fertilizing ability.
朊病毒样蛋白多普蛋白(Doppel)的功能被认为与雄性生育能力有关。在本研究中,评估了绵羊多普蛋白多肽对精子获能和受精的重要性。复性后,在获能过程中向绵羊精子补充不同浓度(40、80或190纳克/毫升)的重组多普蛋白(rDpl)。在实验1中,将解冻后的绵羊精子与不同浓度的rDpl孵育1小时以进行上浮,监测精子活力、浓度、活力、存活率和获能的变化(10次重复)。在实验2中,通过对牛体外成熟卵母细胞进行异源受精,测试上述孵育的上浮后精子的受精能力(n = 423,三次重复)。无论剂量如何,rDpl均可提高(p≤0.03)精子存活率。接受190纳克/毫升rDpl的组中精子个体活力和活力最高(p≤0.04)。补充最高剂量rDpl的精子比对照组(39.1±2.2%)和40纳克/毫升rDpl组(39.8±2.7%)实现了更高(p = 0.02)的受精率(56.0±3.0%)。初步数据表明,多普蛋白可能增强体外精子受精能力。