Neutralization of Vascular Endothelial Growth Factor Antiangiogenic Isoforms Is More Effective Than Treatment with Proangiogenic Isoforms in Stimulating Vascular Development and Follicle Progression in the Perinatal Rat Ovary
Neutralization of Vascular Endothelial Growth Factor Antiangiogenic Isoforms Is More Effective Than Treatment with Proangiogenic Isoforms in Stimulating Vascular Development and Follicle Progression in the Perinatal Rat Ovary
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DOI：
10.1095/biolreprod.109.078097
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发表时间：
2009-11-01
影响因子：
3.6
通讯作者：
Cupp, Andrea S.
中科院分区：
文献类型：
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作者：
Artac, Robin A.;McFee, Renee M.;Cupp, Andrea S.
抑制血管内皮生长因子 A (VEGFA) 信号转导可阻止血管和卵泡发育。由于抗血管生成 VEGFA 异构体被认为可以阻止促血管生成 VEGFA 异构体与其受体结合，因此我们假设促血管生成异构体促进而抗血管生成异构体抑制这些过程。从大鼠卵巢中扩增并测序了抗血管生成亚型 Vegfa_165b 和 Vegfa_189b。 Vegfa_165b 序列与人 VEGFA_165B 具有 90% 同源性。定量 RT-PCR 确定 Vegfa_165b mRNA 在胚胎第 18 天左右更加丰富，但 Vegfa_189b 缺乏明显的丰度模式。抗血管生成 VEGFA 亚型定位于所有卵泡阶段的前颗粒细胞和颗粒细胞以及晚期卵泡的卵泡膜细胞。为了确定 VEGFA 同工型对卵巢发育的影响，将出生后第 3/4 天的大鼠卵巢与 VEGFA_164 或抗血管生成同工型抗体（抗 VEGFAxxxB） 一起培养。用 50 ng/ml VEGFA_164 治疗导致血管密度增加 93%（P < 0.01），并且与对照组相比，治疗的卵巢由较少的原始卵泡（0 期）和更多的发育卵泡（1-4 期）组成（P < 0.04）。与对照组相比，用 5 ng/ml VEGFAxxxB 抗体处理的卵巢血管密度增加了 93% (P < 0.02)，原始和早期初级卵泡（第 1 阶段）较少，初级、移行和次级卵泡（分别为 2、3 和 4 阶段）较多（P < 0.005）。我们得出的结论是，与促血管生成同工型 VEGFA_164 治疗相比，中和抗血管生成 VEGFA 亚型可能是增强围产期大鼠卵巢血管和卵泡发育的更有效机制。
Inhibition of vascular endothelial growth factor A (VEGFA) signal transduction arrests vascular and follicle development. Because antiangiogenic VEGFA isoforms are proposed to block proangiogenic VEGFA isoforms from binding to their receptors, we hypothesized that proangiogenic isoforms promote and antiangiogenic isoforms inhibit these processes. The antiangiogenic isoforms Vegfa_165b and Vegfa_189b were amplified and sequenced from rat ovaries. The Vegfa_165b sequence was 90% homologous to human VEGFA_165B. Quantitative RT-PCR determined that Vegfa_165b mRNA was more abundant around Embryonic Day 18, but Vegfa_189b lacked a distinct pattern of abundance. Antiangiogenic VEGFA isoforms were localized to pregranulosa and granulosa cells of all follicle stages and to theca cells of advanced-stage follicles. To determine the effects of VEGFA isoforms in developing ovaries, Postnatal Day 3/4 rat ovaries were cultured with VEGFA_164 or an antibody to antiangiogenic isoforms (anti-VEGFAxxxB). Treatment with 50 ng/ml of VEGFA_164 resulted in a 93% increase in vascular density (P < 0.01), and treated ovaries were composed of fewer primordial follicles (stage 0) and more developing follicles (stages 1-4) than controls (P < 0.04). Ovaries treated with 5 ng/ml of VEGFAxxxB antibody had a 93% increase in vascular density (P < 0.02), with fewer primordial and early primary follicles (stage 1) and more primary, transitional, and secondary follicles (stages 2, 3, and 4, respectively) compared with controls (P < 0.005). We conclude that neutralization of antiangiogenic VEGFA isoforms may be a more effective mechanism of enhancing vascular and follicular development in perinatal rat ovaries than treatment with the proangiogenic isoform VEGFA_164.