Department of Pathology, Massachusetts General Hospital, Boston, MA, USA.
Reprod Biol Endocrinol. 2010 Aug 16;8:98. doi: 10.1186/1477-7827-8-98.
Ovarian surface epithelial cells are thought to be a precursor cell type for ovarian carcinoma. It has been proposed that an increased rate of ovarian surface epithelial cell proliferation during ovulatory wound repair contributes to the accumulation of genetic changes and cell transformation. The proliferation of ovarian surface epithelial cells during ovulatory wound repair has been studied primarily using immunohistochemical staining of paraffin-embedded ovary sections. However, such analyses require complex reconstruction from serially-cut ovary sections for the visualization and quantification of the cells on the ovarian surface. In order to directly visualize the proliferation and organization of the ovarian surface epithelial cells, we developed a technique for immunohistochemical staining of whole mouse ovaries. Using this method, we analyzed cell proliferation and morphologic changes in mouse ovarian surface epithelial cells during follicle growth and ovulatory wound repair.
Three-week old FVB/N female mice were superovulated by sequential administration of pregnant mare's serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG). Ten hours after hCG administration, mice were given 5-bromo-2-deoxyuridine (BrdU) and euthanized two hours after BrdU administration for ovary isolation. The levels of incorporated BrdU in the ovarian surface epithelial cells were measured by staining paraffin-embedded ovary sections and whole ovaries with the BrdU antibody. Re-epithelialization of the ovarian surface after ovulatory rupture was visualized by immunohistochemical staining with E-cadherin and Keratin 8 in paraffin-embedded ovary sections and whole ovaries.
We determined that active proliferation of ovarian epithelial surface cells primarily occurs during antral follicle formation and, to a lesser extent, in response to an ovulatory wound. We also demonstrated that ovarian surface epithelial cells exhibit a circular organization around the wound site
Whole ovary immunohistochemistry enables efficient and comprehensive three-dimensional visualization of ovarian surface epithelial cells without the need for laborious reconstruction from immunohistochemically-stained serial ovary sections.
卵巢表面上皮细胞被认为是卵巢癌的前体细胞类型。有人提出，在排卵性伤口修复过程中，卵巢表面上皮细胞的增殖速度增加有助于遗传变化和细胞转化的积累。排卵性伤口修复过程中卵巢表面上皮细胞的增殖主要通过石蜡包埋卵巢切片的免疫组织化学染色进行研究。然而，此类分析需要对连续切片的卵巢进行复杂的重建，以实现对卵巢表面细胞的可视化和量化。为了直接观察卵巢表面上皮细胞的增殖和组织，我们开发了一种用于整个小鼠卵巢免疫组织化学染色的技术。使用这种方法，我们分析了卵泡生长和排卵性伤口修复过程中小鼠卵巢表面上皮细胞的细胞增殖和形态变化。
3 周龄的 FVB/N 雌性小鼠通过序贯给予孕马血清促性腺激素（PMSG）和人绒毛膜促性腺激素（hCG）进行超排卵。hCG 给药 10 小时后，给予小鼠 5-溴-2-脱氧尿苷（BrdU），并在 BrdU 给药后 2 小时处死以分离卵巢。通过用 BrdU 抗体对石蜡包埋的卵巢切片和整个卵巢进行染色，测量卵巢表面上皮细胞中掺入的 BrdU 水平。通过在石蜡包埋的卵巢切片和整个卵巢中用 E-钙粘蛋白和角蛋白 8 进行免疫组织化学染色，观察排卵性破裂后卵巢表面的再上皮化。
我们确定卵巢上皮表面细胞的活跃增殖主要发生在窦卵泡形成过程中，并且在较小程度上响应排卵性伤口。我们还表明，卵巢表面上皮细胞在伤口部位周围呈现出圆形组织。
整个卵巢免疫组织化学使我们能够高效且全面地对卵巢表面上皮细胞进行三维可视化，而无需对免疫组织化学染色的连续卵巢切片进行繁琐的重建。
