The Lymphatic System in the Primate corpus Luteum

描述（由申请人提供）：淋巴系统由薄壁血管和淋巴结组成，这些血管和淋巴结收集富含蛋白质的液体（即淋巴）并将其返回到血液中。淋巴作为循环系统的一部分，对维持体液平衡和组织稳态起着重要作用。最近的进展提供了对淋巴管在胚胎发生和癌症转移过程中的调节和作用的见解，但关于它们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生理的一个很好的器官，因为黄体等配子体/内分泌组织的周期性和暂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，还没有关于灵长类动物淋巴系统的报道。这类研究是必要的，因为有人提出淋巴管的范围和作用与淋巴管大小正相关，即典型实验室啮齿动物的小淋巴管几乎没有淋巴管。因此，我们提出研究来验证淋巴系统在灵长类动物中存在的假设，以及淋巴系统的发育与淋巴管生成有关。以恒河猴为灵长类动物模型，研究设计：(a)目的1 -研究灵长类动物卵巢中淋巴管的存在、定位和动态，特别是在月经周期的整个黄体生命周期中淋巴管的存在、定位和动态。新发现的淋巴内皮细胞（LECs）标志物，如PROX-1、LYVE-1、podoplanin和VEGFR-3，将通过免疫细胞化学验证，并通过图像分析定量LECs的数量和淋巴面积；(b)目的2 -分析新型淋巴管生成因子血管内皮生长因子(VEGF)- c和- d及其受体VEGFR-3在灵长类动物CL中的表达。采用Real time PCR、western blotting、原位杂交和免疫细胞化学方法定量测定月经周期卵巢/CL中mRNA和蛋白的表达，并定位表达到特定细胞类型。这将有助于我们理解灵长类动物卵巢淋巴系统的结构功能，并为进一步研究灵长类动物卵巢淋巴系统在正常月经周期和卵巢疾病（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理论基础。

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.