CLARITY reveals dynamics of ovarian follicular architecture and vasculature in three-dimensions.
CLARITY reveals dynamics of ovarian follicular architecture and vasculature in three-dimensions.
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CLARITY 揭示了三维卵泡结构和脉管系统的动态
DOI：
10.1038/srep44810
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发表时间：
2017-03-23
影响因子：
4.6
通讯作者：
Hsueh AJ
中科院分区：
文献类型：
--
作者：
Feng Y;Cui P;Lu X;Hsueh B;Möller Billig F;Zarnescu Yanez L;Tomer R;Boerboom D;Carmeliet P;Deisseroth K;Hsueh AJ
器官内异质细胞器和细胞类型的最佳分布对于生理过程是必不可少的。卵巢特有的经尿道调节的卵泡发生、排卵、黄体形成/消退和相关的脉管系统变化导致每个生殖周期中的组织重塑。我们采用免疫组织化学方法和标记物免疫染色，在完整的卵巢中识别单个卵泡和黄体。监测卵泡群体的寿命变化显示，总卵泡和高级卵泡的百分比随年龄而下降。从原始卵泡到排卵前卵泡发育的特征是体积增加3 × 105倍，圆形度降低，同一阶段卵泡的聚集减少。卵泡-血管系统关系图的构建表明卵泡周围血管系统和分支的年龄和促性腺激素依赖性增加。血管内皮生长因子A（VEGFA）启动子中缺氧反应元件缺失的杂合突变小鼠表现出卵巢血管缺陷和排卵反应降低。单侧囊内注射血管内皮生长因子受体抑制剂阿西替尼，延缓了治疗卵巢中与排卵缺陷相关的新血管生成。我们的方法揭示了卵巢结构的独特特征和血管系统在组织卵泡中的重要作用，以便将来对正常和患病的人类卵巢进行研究。类似的方法也可以揭示胚胎发育和肿瘤发生过程中新血管生成的作用。
Optimal distribution of heterogeneous organelles and cell types within an organ is essential for physiological processes. Unique for the ovary, hormonally regulated folliculogenesis, ovulation, luteal formation/regression and associated vasculature changes lead to tissue remodeling during each reproductive cycle. Using the CLARITY approach and marker immunostaining, we identified individual follicles and corpora lutea in intact ovaries. Monitoring lifetime changes in follicle populations showed age-dependent decreases in total follicles and percentages of advanced follicles. Follicle development from primordial to preovulatory stage was characterized by 3 × 105-fold increases in volume, decreases in roundness, and decreased clustering of same stage follicles. Construction of follicle-vasculature relationship maps indicated age- and gonadotropin-dependent increases in vasculature and branching surrounding follicles. Heterozygous mutant mice with deletion of hypoxia-response element in the vascular endothelial growth factor A (VEGFA) promoter showed defective ovarian vasculature and decreased ovulatory responses. Unilateral intrabursal injection of axitinib, an inhibitor of VEGF receptors, retarded neo-angiogenesis that was associated with defective ovulation in treated ovaries. Our approach uncovers unique features of ovarian architecture and essential roles of vasculature in organizing follicles to allow future studies on normal and diseased human ovaries. Similar approaches could also reveal roles of neo-angiogenesis during embryonic development and tumorigenesis.