Both VEGF and its receptor proteins were localized by immunostaining technique in the luminal epithelium, smooth muscle cells and blood vessels within the oviduct. Moreover, by means of semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) techniques, it has been demonstrated that mRNA of VEGF and its receptors in both healthy and diseased oviduct is expressed preferentially at the time and place where the amount of oviduct fluid is prominent. This supports the notion that VEGF may be a regulator of oviductal secretion. This thesis has consistently demonstrated a modulation pattern of flt-1 expression that is similar to its ligand VEGF in both physiological and pathological conditions. This suggests that flt-1 may be the main receptor responsible for the action of VEGF in the oviduct. As illustrated in both the in-vivo and in-vitro models, the expression of VEGF and flt-1 in the human oviduct is stimulated directly by gonadotropins without the influence of ovarian sex hormones. / Increased knowledge on the regulatory mechanisms of oviductal fluid formation, the first environment that human embryos are exposed to, will be valuable from the clinical management point of view. / Oviductal fluid is a complex mixture of plasma-derived constituents and proteins synthesized by the oviduct epithelium. It has been postulated that vascular endothelial growth factor (VEGF), a known permeability promoter, may be an important regulator of oviductal fluid secretion by stimulating vascular permeability and so serum transudation. However, little is known about the expression of VEGF in the human oviduct. This thesis investigated the modulation of VEGF and its receptors (flt-1 and KDR) in the healthy oviducts, from fertile women undergoing tubal sterilization for unwanted fertility or hysterectomy for benign gynecological conditions, as well as in the hydrosalpinges from sterile women undergoing salpingectomy before the treatment of in-vitro fertilization and embryo transfer. / Lam Po Mui. / Adviser: Christopher J. Haines. / Source: Dissertation Abstracts International, Volume: 73-01, Section: B, page: . / Thesis (M.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 147-179). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 5 uppsatser.

Diss. (sammanfattning) Stockholm : Karol inst., 2004. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 4 uppsatser.

published_or_final_version / abstract / Dentistry / Doctoral / Doctor of Philosophy

published_or_final_version / abstract / Obstetrics and Gynaecology / Doctoral / Doctor of Philosophy

Yeung Lam. / Thesis submitted in: December 2003. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 134-150). / Abstracts in English and Chinese. / ABSTRACT --- p.I / 摘要 --- p.III / ACKNOWLEDGMENT --- p.V / Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- General review of angiogenesis --- p.1 / Chapter 1.2 --- Vascular endothelial growth factors (VEGFs) --- p.2 / Chapter 1.2.1 --- VEGF-A --- p.2 / Chapter 1.2.2 --- P1GF --- p.4 / Chapter 1.2.3 --- VEGF-B --- p.5 / Chapter 1.2.4 --- VEGF-C and VEGF-D --- p.6 / Chapter 1.3 --- VEGF receptors (VEGFRs) --- p.9 / Chapter 1.3.1 --- VEGFR-1 (or flt-1) --- p.9 / Chapter 1.3.2 --- VEGFR-2 ( or flk-1) --- p.10 / Chapter 1.3.3 --- VEGFR-3 ( or flt-4) --- p.11 / Chapter 1.4 --- Hormonal regulation of VEGFs by LH/hCG --- p.14 / Chapter 1.5 --- General review of the testis --- p.17 / Chapter 1.5.1 --- Structure and function of the testis --- p.17 / Chapter 1.5.2 --- Testicular vasculature --- p.18 / Chapter 1.5.3 --- Testicular angiogenesis --- p.19 / Chapter 1.6 --- Localization of VEGF and VEGF receptors in the testis --- p.20 / Chapter 1.7 --- Aims of the present study --- p.21 / Chapter 2. --- Materials and methods --- p.23 / Chapter 2.1 --- Animals --- p.23 / Chapter 2.1.1 --- Depletion of Leydig cell --- p.23 / Chapter 2.1.2 --- Suppression of Leydig cell and stimulation by hCG --- p.24 / Chapter 2.1.3 --- Collection of tissue --- p.25 / Chapter 2.2 --- Preparation of primary cells from rat testes --- p.27 / Chapter 2.2.1 --- Sertoli cell preparation --- p.27 / Chapter 2.2.2 --- Germ cell preparation --- p.29 / Chapter 2.2.3 --- Interstitial cell and Leydig cell preparation --- p.30 / Chapter 2.3 --- Cell cultures --- p.32 / Chapter 2.3.1 --- Reagents and cell lines --- p.32 / Chapter 2.3.2 --- "Mouse Leydig cell line, TM3 and Sertoli cell line, TM4" --- p.33 / Chapter 2.3.3 --- "Mouse tumor Leydig cell line, MLTC-1" --- p.34 / Chapter 2.3.4 --- "Rat tumor Leydig cell line, R2C" --- p.34 / Chapter 2.3.5 --- "Rat tumor Leydig cell line, LC540" --- p.35 / Chapter 2.4 --- Reverse-transcription polymerase chain reaction (RT-PCR) and semi-quantitative RT-PCR --- p.35 / Chapter 2.4.1 --- Extraction of total RNA --- p.35 / Chapter 2.4.2 --- Quantitation of total RNA --- p.37 / Chapter 2.4.3 --- RT-PCR --- p.37 / Chapter 2.4.4 --- Purification and authentication of PCR products --- p.47 / Chapter 2.5 --- Immunohistochemical staining --- p.48 / Chapter 2.5.1 --- Perfusion and processing of testes for histological sections --- p.48 / Chapter 2.5.2 --- Immunohistochemical staining of tissue sections --- p.50 / Chapter 2.6 --- Western immunoblotting --- p.52 / Chapter 2.6.1 --- Extraction and quantitation of total protein --- p.52 / Chapter 2.6.2 --- SDS-PAGE --- p.53 / Chapter 2.6.3 --- Immunoblotting --- p.55 / Chapter 2.7 --- Statistical analyses --- p.57 / Chapter 3. --- Results --- p.58 / Chapter 3.1 --- Expression and localization of VEGFs in the rat testis --- p.58 / Chapter 3.1.1 --- VEGF-A --- p.58 / Chapter 3.1.2 --- VEGF-B --- p.64 / Chapter 3.1.3 --- VEGF-C --- p.69 / Chapter 3.1.4 --- VEGF-D --- p.73 / Chapter 3.1.5 --- P1GF --- p.77 / Chapter 3.2 --- Effect of Leydig cell depletion on VEGFs expression in the rat testis --- p.81 / Chapter 3.2.1 --- Effect on VEGF-A --- p.81 / Chapter 3.2.2 --- Effect on VEGF-B --- p.82 / Chapter 3.2.3 --- Effect on VEGF-C --- p.88 / Chapter 3.2.4 --- Effect on VEGF-D --- p.91 / Chapter 3.2.5 --- Effect on P1GF --- p.94 / Chapter 3.3 --- Effect of Leydig cell suppression and hCG stimulation on VEGFs expression in the rat testis --- p.97 / Chapter 3.3.1 --- Effect on VEGF-A --- p.97 / Chapter 3.3.2 --- Effect on VEGF-B --- p.107 / Chapter 3.3.3 --- Effect on VEGF-C --- p.113 / Chapter 3.3.4 --- Effect on VEGF-D --- p.119 / Chapter 4. --- Discussion --- p.126 / Chapter 5. --- References --- p.134

Thesis (Ph. D.)--University of Hong Kong, 2007. / Title proper from title frame. Also available in printed format.

Thesis (Ph. D.)--University of Hong Kong, 2006. / Title proper from title frame. Also available in printed format.

Choroidal neovascularization (CNV) is a leading threat to severe vision loss, particularly in patients with age-related macular degeneration (AMD). In CNV, newly formed blood vessels sprout from the choroid to the sub-retinal space, where leakage and bleeding of the abnormal vessels lead to photoreceptor death and subsequent vision loss. It is believed that CNV is mediated by growth factors (e.g. vascular endothelial growth factor {VEGF}) produced by the retinal pigment epithelium (RPE) under pathological states (e.g. hypoxia). Current treatments for CNV aiming at countering VEGF only help decrease leakage and inhibit formation of CNV, but none of them is curative and the recurrence rate remains high. In order to find other more powerful potential therapeutic targets, the regulations of VEGF signaling in the pathophysiology of CNV is the focus of numerous translational investigations. Previously, Hypoxia-inducible factor-1 (HIF-1), a crucial transcriptional factor in response to hypoxia, is identified as the master transcriptional factor controlling VEGF expression in the RPE promoting CNV. Alpha-enolase (ENO1), a key glycolytic enzyme, is known to be over expressed in several types of carcinomas also under the regulation of HIF-1. ENO1 has been reported to be closely associated with cancer progression, angiogenesis, and venous invasion. The molecular events of ENO1 in the pathogenesis of promoting angiogenesis are of interest but still barely understood. Recently, the association of ENO1 antibodies with retina has been seen in patients with AMD. We hypothesize that ENO1 expression in the RPE may play a role in the development of CNV, participating in the regulation of VEGF. Hypoxia is an important pathological condition in the formation of CNV. Here, we first determined ENO1 expression and cell death in a human RPE cell line, ARPE-19, under cobalt (II) chloride (CoCl2)-induced hypoxia or anoxia (95% N2, 5% CO2). To further investigate the regulation of ENO1 in CNV, HIF-1α-diminished RPE cells were generated using small interfering RNA (siRNA) and the change of ENO1 expression in response to hypoxic injury was determined. Upon 24 hr of treatment with CoCl2-induced hypoxia or anoxia, the expression of ENO1 and VEGF increased significantly along with HIF-1α in ARPE-19 cells, both of which could in turn be significantly down-regulated by HIF-1α siRNA. Interestingly, cell death remained low in ARPE-19 cells, even after 24 hr of CoCl2-induced hypoxia or anoxia. To further study the role of ENO1 in CNV, we started by investigating the relationship between ENO1 and VEGF. SiRNA was used to knock down the expression of ENO1 in ARPE-19 cells. Upon transfection with the siRNA, ENO1 expression was successfully down-regulated when treated with CoCl2-induced hypoxia. However, VEGF secretions from the ENO1-diminished ARPE-19 cells under CoCl2-induced hypoxia remained unchanged. Double knockdown of ENO1 together with HIF-1α by siRNA also did not help to further suppress VEGF secretion in the hypoxic ARPE-19 cells. Hence, ENO1 was demonstrated to be activated and up-regulated by HIF-1 in RPE cells responding to hypoxia, suggesting a potential role of ENO1 in favoring the formation of CNV, but not through influencing VEGF secretion. / published_or_final_version / Ophthalmology / Master / Master of Philosophy