The Lymphatic System in the Primate corpus Luteum

描述(申请人提供)：淋巴系统由薄壁血管和淋巴结组成，收集富含蛋白质的液体(即淋巴)并将其返回血液。淋巴管作为循环系统的一部分，对维持体液平衡和组织内环境平衡非常重要。最近的进展为淋巴管在胚胎发生和癌症转移中的调节和作用提供了深入的见解，但关于淋巴管在健康成人组织中的发育(淋巴管生成)、维持或破坏(淋巴管溶解)的信息很少。卵巢应该是研究淋巴管生理学的一个很好的器官，因为配子/内分泌组织(如黄体)的周期性和临时性形成与血流和血管通透性的显著变化有关。到目前为止，还没有关于灵长类CL淋巴管的报道。这类研究是必要的，因为有人提出淋巴管的范围和作用与CL大小呈正相关，即典型实验室啮齿动物的小CL几乎没有淋巴管。因此，有研究提出了一种假设，即灵长类CL中存在淋巴系统，并且CL的发育与淋巴管的生成有关。使用恒河猴作为灵长类动物模型，设计了以下研究：(A)目的1-研究灵长类卵巢中淋巴管的存在、定位和动力学，特别是在月经周期的整个黄体周期中淋巴管在CL中的存在、定位和动力学。新发现的淋巴管内皮细胞标志物，如Prox-1、Lyve-1、podoplan in和VEGFR-3，将通过免疫细胞化学进行验证，并通过图像分析来定量淋巴管内皮细胞的数量和淋巴面积；(B)目的2-分析新的淋巴管生成因子血管内皮生长因子C和-D及其受体VEGFR-3在灵长类CL中的表达。实时定量聚合酶链式反应和免疫印迹、原位杂交和免疫细胞化学将用于定量检测基因和蛋白的表达，并在月经周期中定位于卵巢/卵巢中特定的细胞类型。这一认识将有助于我们理解灵长类CL的结构和功能，并为进一步研究淋巴系统在正常月经周期中灵长类卵巢和卵巢疾病(如卵巢过度刺激综合征或卵巢囊肿)中的调节和作用提供坚实的基础。

DESCRIPTION (provided by applicant): The lymphatic system consists of thin-walled vessels and lymph nodes that collect protein-rich fluid (i.e. lymph) and return it to the blood stream. As a part of the circulatory system, the lymphatics are important for maintaining fluid balance and tissue homeostasis. Recent advances provided insight into the regulation and role of the lymphatics during embryogenesis and cancer metastasis, but there is little information on their development (lymphangiogenesis), maintenance, or destruction (lymphangiolysis) in healthy, adult tissues. The ovary should be an excellent organ for studying the physiology of the lymphatics, since the cyclic and temporary formation of gametogenic/endocrine tissues such as the corpus luteum (CL) is associated with remarkable changes in blood flow and vascular permeability. To date, there are no reports on the lymphatics in the primate CL. Such studies are needed since it is proposed that the extent and role of the lymphatics is positively correlated with CL size, i.e., the small CL of typical laboratory rodents is virtually devoid of lymphatics. Therefore, studies are proposed to test the hypothesis that the lymphatic system exists in the primate CL and that CL development is associated with lymphangiogenesis. Using the rhesus monkey as a primate model, studies were designed: (a) Aim No.1 - To investigate the presence, localization, and dynamics of lymphatic vessels in the primate ovary, specifically in the CL throughout the luteal lifespan of the menstrual cycle. Newly - discovered markers for lymphatic endothelial cells (LECs), e.g., PROX-1, LYVE-1, podoplanin, and VEGFR-3, will be validated by immunocytochemistry, and the numbers of LECs and lymphatic area quantitated by image analysis; (b) Aim No.2 - To analyze the expression of novel lymphangiogenic factors, vascular endothelial growth factor (VEGF)-C and -D, and their receptor VEGFR-3 in the primate CL. Real time PCR and western blotting, plus in situ hybridization and immunocytochemistry will be employed to quantitate mRNA and protein expression, plus localize expression to specific cell types in the ovary/CL during the menstrual cycle. This knowledge will contribute to our understanding of the structure-function of the primate CL, and should provide a solid rationale for further studies on the regulation and role of the lymphatic system in the primate ovary during the normal menstrual cycle and in ovarian disorders, such as ovarian hyperstimulation syndrome or ovarian cysts.