McNatty Kenneth P, Hudson Norma L, Whiting Lynda, Reader Karen L, Lun Stan, Western Andrea, Heath Derek A, Smith Peter, Moore Lloyd G, Juengel Jennifer L
AgResearch, Wallaceville Animal Research Centre, Upper Hutt 6007, New Zealand.
Biol Reprod. 2007 Apr;76(4):552-60. doi: 10.1095/biolreprod.106.054361. Epub 2006 Nov 8.
The aims of these studies were to determine the abilities of antisera against different regions of ovine bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9) to inhibit ovarian follicular activity, estrus (mating), and ovulation in sheep. The 9-15-mer peptides were conjugated to keyhole limpet hemocyanin (KLH) and used to generate antibodies against the flexible N-terminal regions of the mature protein as well as against regions in which dimerization of the protein or interaction with a type 1 BMP or a type 2 TGFB or BMP receptor was predicted to occur. Ewes (n = 10 per treatment group) were vaccinated with KLH or the KLH-BMP15 (n = 9 different peptides) or KLH-GDF9 (n = 10) peptides in Freund adjuvant at five consecutive monthly intervals. Overall, antisera generated against peptides that corresponded to amino acid residues 1-15 of the N-terminus of the BMP15 or GDF9 mature protein or GDF9 amino acid residues 21-34 were the most potent at inhibiting ovulation following primary and single booster vaccination. Several other BMP15 (8/9) or GDF9 (6/10) treatment groups, but not KLH alone, also produced significant reductions in the numbers of animals that ovulated, although 2, 3 or 4 booster vaccinations were required. Anovulation was commonly associated with the inhibition of normal ovarian follicular development and anestrus. The in vitro neutralization studies with IgG from the BMP15 or GDF9 immunized ewes showed that the mean inhibition of BMP15 plus GDF9 stimulation of (3)H-thymidine uptake by rat granulosa cells was approximately 70% for animals without corpora lutea (CL), whereas for animals with one to three CL or more than three CL, the inhibition was 24%-33% or 27%-42%, respectively. In summary, these data suggest that reagents that block the biological actions of BMP15 or GDF9 at their N-termini have potential as contraceptives or sterilizing agents.
这些研究的目的是确定抗绵羊骨形态发生蛋白15（BMP15）和生长分化因子9（GDF9）不同区域的抗血清抑制绵羊卵巢卵泡活性、发情（交配）和排卵的能力。将9 - 15个氨基酸的肽与钥孔血蓝蛋白（KLH）偶联，用于产生针对成熟蛋白柔性N端区域以及预测蛋白二聚化或与1型BMP或2型TGFB或BMP受体相互作用区域的抗体。母羊（每个治疗组n = 10）在连续五个月的每月间隔时间用KLH或KLH - BMP15（n = 9种不同肽）或KLH - GDF9（n = 10）肽在弗氏佐剂中进行免疫接种。总体而言，针对与BMP15或GDF9成熟蛋白N端1 - 15个氨基酸残基或GDF9 21 - 34个氨基酸残基相对应的肽产生的抗血清在初次和单次加强免疫后抑制排卵的效果最强。其他几个BMP15（8/9）或GDF9（6/10）治疗组，但不包括单独的KLH组，也使排卵动物数量显著减少，尽管需要进行2、3或4次加强免疫。无排卵通常与正常卵巢卵泡发育抑制和发情停止有关。用来自BMP15或GDF9免疫母羊的IgG进行的体外中和研究表明，对于无黄体（CL）的动物，BMP15加GDF9刺激大鼠颗粒细胞摄取（3）H - 胸腺嘧啶的平均抑制率约为70%，而对于有1至3个CL或超过3个CL的动物，抑制率分别为24% - 33%或27% - 42%。总之，这些数据表明在N端阻断BMP15或GDF9生物学作用的试剂有作为避孕药或绝育剂的潜力。
