Angiogenesis in chicken chorioallantoic membrane under hypoxic incubations

2017 ◽  
Vol 67 (3-4) ◽  
pp. 251-261 ◽  
Author(s):  
Hongliang Zhang ◽  
Peng Shang ◽  
Yawen Zhang ◽  
Ying Zhang ◽  
Xiaolong Tian ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Blood Vessels ◽  
Quantitative Pcr ◽  
Early Stage ◽  
Chorioallantoic Membrane ◽  
Developmental Timing ◽  
Vascular Density ◽  
Hypoxic Conditions ◽  
Density Index ◽  
Chicken Chorioallantoic Membrane

Under hypoxic conditions, angiogenesis in the chorioallantoic membrane exhibits a sensitive response depending on the developmental timing and intensity and duration of the hypoxia. Furthermore, the effects of hypoxia on vascularization in the chorioallantoic membrane are controversial. In this study, we used microscopy to determine the vascular density index of chicken embryonic chorioallantoic membrane and quantitative PCR to examine the expression of the HIF-1α and VEGFA genes. Two levels of hypoxia (15% and 13% O2) and three durations (days 0–10, 11–18, and 0–18) were applied. The results showed that 13% O2 incubation restrained angiogenesis in the chorioallantoic membrane at an early stage of embryonic development as seen on day 6, but incubation under 13% or 15% O2 efficiently stimulated vascularization of the chorioallantoic membrane as seen on days 16 and 18. Notably, 13% O2 incubation caused visible curling of the blood vessels in the chorioallantoic membrane on day 18. The increased vascular density index under hypoxia was accompanied by an increase in the expression of VEGFA and HIF-1α. Curled blood vessels were observed on day 18 under 13% O2 incubation; however, the curling was not a result of VEGFA overexpression. Thus, the present study helps in elucidating vascularization of the chorioallantoic membrane under hypoxic conditions.

scholarly journals An antagonist of integrin alpha v beta 3 prevents maturation of blood vessels during embryonic neovascularization

1995 ◽  
Vol 108 (7) ◽  
pp. 2655-2661 ◽  
Author(s):  
C.J. Drake ◽  
D.A. Cheresh ◽  
C.D. Little
Keyword(s):  
Experimental Data ◽  
Monoclonal Antibody ◽  
Blood Vessels ◽  
Chorioallantoic Membrane ◽  
Normal Pattern ◽  
Basal Surface ◽  
Lumen Formation ◽  
Vessel Development ◽  
Alpha V Beta 3 ◽  
Chicken Chorioallantoic Membrane

Experimental data in this study demonstrate that integrin alpha v beta 3 is fundamentally involved in the maturation of blood vessels during embryonic neovascularization (vasculogenesis). Integrin alpha v beta 3 was specifically expressed on the surface of angioblasts during vessel development in quail embryos and vitronectin, a ligand for alpha v beta 3, localized to the basal surface of these cells. More importantly, microinjection of the anti-alpha v beta 3 monoclonal antibody, LM609, disrupted the normal pattern of vascular development. After exposure to LM609 the angioblasts in experimental embryos appeared as clusters of rounded cells lacking normal cellular protrusions. This led to disruption of lumen formation and abnormal vessel patterning. These findings demonstrate that during vasculogenesis ligation of integrin alpha v beta 3 on the surface of primordial endothelial cells is critical for the differentiation and maturation of blood vessels. Similar studies on chicken chorioallantoic membrane showed that LM609 blocks angiogenesis. Together the two studies suggest that integrin alpha v beta 3 plays a role in neovascularization of tissues.

scholarly journals Circulation Index as a Quantitative Indicator of Angiogenesis in Chorioallantoic Membrane of Chicken Broilers

2018 ◽  
Vol 67 (2) ◽  
pp. 164-170
Author(s):  
Zoran Ružić ◽  
Zdenko Kanački ◽  
Dragan Žikić ◽  
Gordana Ušćebrka ◽  
Jovan Mirčeta
Keyword(s):  
Embryonic Development ◽  
Blood Vessels ◽  
Chorioallantoic Membrane ◽  
New Method ◽  
In Vivo Studies ◽  
Circulation Index ◽  
Fast Method ◽  
Angiogenic Response ◽  
Embryonic Life

Summary Chorioallantoic membrane (CAM) is an extraembryonic membrane very frequently used for in vivo studies in various researches. Since researches require a fast method for quantifying the CAM angiogenic response, there is a need to develop a new precise and unbiased method of quantification of angiogenesis in CAM, which would be easy to perform and suitable for analysis of a large number of samples. The objective of this paper is to apply a new method of quantification of angiogenesis in investigation of the development of blood vessels in the CAM, in particular days of embryonic life considered essential for CAM development. The present research included 75 fertilized eggs of heavy hybrid Ross 308. CAM sampling for stereological analyses was in key phases of embryonic development, namely on the 12th, 15th and 19th day. The results of the present investigation show that the increase in embryonic age results in increase in circulation index, which is also an indicator of angiogenic processes developing in CAM. The lowest value of circulation index (0.1952) was recorded on the first sampling day (E12), while the highest value (0.2666) was recorded on the last sampling day (E19). This method may be applied in researching different factors which affect angiogenesis in CAM.

scholarly journals Transforming growth factor beta 1-induced changes in cell migration, proliferation, and angiogenesis in the chicken chorioallantoic membrane.

1990 ◽  
Vol 111 (2) ◽  
pp. 731-741 ◽  
Author(s):  
E Y Yang ◽  
H L Moses
Keyword(s):  
Endothelial Cell ◽  
Blood Vessels ◽  
Cell Density ◽  
Chorioallantoic Membrane ◽  
Cell Types ◽  
Cellular Responses ◽  
Endothelial Cell Proliferation ◽  
Tgf Beta ◽  
Inhibition Of Proliferation ◽  
Chicken Chorioallantoic Membrane

Application of TGF beta 1 (10-100 ng) to the chicken chorioallantoic membrane (CAM) for 72 h resulted in a dose-dependent, gross angiogenic response. The vascular effects induced by TGF beta 1 were qualitatively different than those induced by maximal doses of basic FGF (bFGF) (500 ng). While TGF beta 1 induced the formation of large blood vessels by 72 h, bFGF induced primarily small blood vessels. Histologic analysis revealed that TGF beta 1 stimulated pleiotropic cellular responses in the CAM. Increases in fibroblast and epithelial cell density in the area of TGF beta 1 delivery were observed as early as 4 h after TGF beta 1 treatment. By 8 h, these cell types also demonstrated altered morphology and marked inhibition of proliferation as evidenced by 3H-thymidine labeling. Thus, the TGF beta 1-stimulated accumulation of these cell types was the result of cellular chemotaxis from peripheral areas into the area of TGF beta 1 delivery. Microscopic angiogenesis in the form of capillary sprouts and increased endothelial cell density first became evident at 16 h. By 24 h, capillary cords appeared within the mesenchyme of the CAM, extending towards the point of TGF beta 1 delivery. 3H-thymidine labeling revealed that the growth of these capillary cords was due to endothelial cell proliferation. Finally, perivascular mononuclear inflammation did not become evident until 48 h of treatment, and its presence correlated spatially and temporally with the gross and histological remodelling of newly formed capillary cords into larger blood vessels. In summary, these data suggest that, in the chicken CAM, TGF beta 1 initiates a sequence of cellular responses that results in growth inhibition, cellular accumulation through migration, and microvascular angiogenesis.

scholarly journals Expression of SPARC during development of the chicken chorioallantoic membrane: evidence for regulated proteolysis in vivo.

1995 ◽  
Vol 6 (3) ◽  
pp. 327-343 ◽  
Author(s):  
M L Iruela-Arispe ◽  
T F Lane ◽  
D Redmond ◽  
M Reilly ◽  
R P Bolender ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Blood Vessels ◽  
Chorioallantoic Membrane ◽  
Focal Adhesions ◽  
Protein Accumulation ◽  
Dependent Manner ◽  
Plasmin Activity ◽  
Specific Expression ◽  
Chicken Chorioallantoic Membrane

SPARC is a secreted glycoprotein that has been shown to disrupt focal adhesions and to regulate the proliferation of endothelial cells in vitro. Moreover, peptides resulting from the proteolysis of SPARC exhibit angiogenic activity. Here we describe the temporal synthesis, turnover, and angiogenic potential of SPARC in the chicken chorioallantoic membrane. Confocal immunofluorescence microscopy revealed specific expression of SPARC protein in endothelial cells, and significantly higher levels of SPARC were observed in smaller newly formed blood vessels in comparison to larger, developmentally older vessels. SPARC mRNA was detected at the earliest stages of chorioallantoic membrane morphogenesis and reached maximal levels at day 13 of embryonic development. Interestingly, steady-state levels of SPARC mRNA did not correlate directly with protein accumulation; moreover, the protein appeared to undergo limited degradation during days 10-15. Incubation of [125I]-SPARC with chorioallantoic membranes of different developmental ages confirmed that extracellular proteolysis occurred during days 9-15, but not at later stages (e.g., days 17-21). Comparison of peptides produced by incubation with chorioallantoic membranes with those generated by plasmin showed an identical pattern of proteolysis. Plasmin activity was present throughout development, and in situ zymography identified sites of plasminogen activator activity that corresponded to areas exhibiting high levels of SPARC expression. Synthetic peptides from a plasmin-sensitive region of SPARC, between amino acids 113-130, stimulated angiogenesis in the chorioallantoic membrane in a dose-dependent manner; in contrast, intact SPARC was inactive in similar assays. We have shown that SPARC is expressed in endothelial cells of newly formed blood vessels in a manner that is both temporally and spatially restricted. Between days 9 and 15 of chorioallantoic membrane development, the protein undergoes proteolytic cleavage that is mediated, in part, by plasmin. SPARC peptides released specifically by plasmin induce angiogenesis in vivo. We therefore propose that SPARC acts as an intrinsic regulator of angiogenesis in vivo.

scholarly journals An angiogenesis platform using a cubic artificial eggshell with patterned blood vessels on chicken chorioallantoic membrane

PLoS ONE ◽  
2017 ◽  
Vol 12 (4) ◽  
pp. e0175595 ◽  
Author(s):  
Wenjing Huang ◽  
Makoto Itayama ◽  
Fumihito Arai ◽  
Katsuko S. Furukawa ◽  
Takashi Ushida ◽  
...  
Keyword(s):  
Blood Vessels ◽  
Chorioallantoic Membrane ◽  
Chicken Chorioallantoic Membrane

Review of Various Tasks Performed in the Preprocessing Phase of a Diabetic Retinopathy Diagnosis System

2020 ◽  
Vol 16 (4) ◽  
pp. 397-426
Author(s):  
Muhammad Nadeem Ashraf ◽  
Muhammad Hussain ◽  
Zulfiqar Habib
Keyword(s):  
Diabetic Retinopathy ◽  
Blood Vessels ◽  
Early Stage ◽  
Spherical Shape ◽  
Cost Effective ◽  
Lesion Detection ◽  
Diabetic Patients ◽  
Cad System ◽  
Non Invasive ◽  
Optic Discs

Diabetic Retinopathy (DR) is a major cause of blindness in diabetic patients. The increasing population of diabetic patients and difficulty to diagnose it at an early stage are limiting the screening capabilities of manual diagnosis by ophthalmologists. Color fundus images are widely used to detect DR lesions due to their comfortable, cost-effective and non-invasive acquisition procedure. Computer Aided Diagnosis (CAD) of DR based on these images can assist ophthalmologists and help in saving many sight years of diabetic patients. In a CAD system, preprocessing is a crucial phase, which significantly affects its performance. Commonly used preprocessing operations are the enhancement of poor contrast, balancing the illumination imbalance due to the spherical shape of a retina, noise reduction, image resizing to support multi-resolution, color normalization, extraction of a field of view (FOV), etc. Also, the presence of blood vessels and optic discs makes the lesion detection more challenging because these two artifacts exhibit specific attributes, which are similar to those of DR lesions. Preprocessing operations can be broadly divided into three categories: 1) fixing the native defects, 2) segmentation of blood vessels, and 3) localization and segmentation of optic discs. This paper presents a review of the state-of-the-art preprocessing techniques related to three categories of operations, highlighting their significant aspects and limitations. The survey is concluded with the most effective preprocessing methods, which have been shown to improve the accuracy and efficiency of the CAD systems.

scholarly journals N-acetyl-L-cysteine Improves the Developmental Competence of Bovine Oocytes and Embryos Cultured In Vitro by Attenuating Oxidative Damage and Apoptosis

Antioxidants ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 860
Author(s):  
Wu-Sheng Sun ◽  
Hoon Jang ◽  
Mi-Ryung Park ◽  
Keon Bong Oh ◽  
Haesun Lee ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Early Stage ◽  
Developmental Competence ◽  
Beneficial Effects ◽  
Developmental Rates ◽  
A Cell ◽  
Bovine Oocytes ◽  
Dose Dependent

Oxidative stress has been suggested to negatively affect oocyte and embryo quality and developmental competence, resulting in failure to reach full term. In this study, we investigated the effect of N-acetyl-L-cysteine (NAC), a cell-permeating antioxidant, on developmental competence and the quality of oocytes and embryos upon supplementation (0.1–10 mM) in maturation and culture medium in vitro using slaughterhouse-derived oocytes and embryos. The results show that treating oocytes with 1.0 mM NAC for 8 h during in vitro maturation attenuated the intracellular reactive oxygen species (ROS) (p < 0.05) and upregulated intracellular glutathione levels (p < 0.01) in oocytes. Interestingly, we found that NAC affects early embryonic development, not only in a dose-dependent, but also in a stage-specific, manner. Significantly (p < 0.05) decreased cleavage rates (90.25% vs. 81.46%) were observed during the early stage (days 0–2), while significantly (p < 0.05) increased developmental rates (38.20% vs. 44.46%) were observed during the later stage (from day 3) of embryonic development. In particular, NAC supplementation decreased the proportion of apoptotic blastomeres significantly (p < 0.05), resulting in enhanced hatching capability and developmental rates during the in vitro culture of embryos. Taken together, our results suggest that NAC supplementation has beneficial effects on bovine oocytes and embryos through the prevention of apoptosis and the elimination of oxygen free radicals during maturation and culture in vitro.

scholarly journals PATH-04. THE BLOOD-BRAIN BARRIER IN DIFFUSE MIDLINE GLIOMA AND ITS IMPLICATIONS FOR DRUG DELIVERY

2020 ◽  
Vol 22 (Supplement_2) ◽  
pp. ii164-ii164
Author(s):  
Rianne Haumann ◽  
Fatma El-Khouly ◽  
Marjolein Breur ◽  
Sophie Veldhuijzen van Zanten ◽  
Gertjan Kaspers ◽  
...  
Keyword(s):  
Drug Delivery ◽  
Blood Vessels ◽  
Blood Brain Barrier ◽  
Brain Barrier ◽  
Vascular Density ◽  
Zonula Occludens ◽  
Blood Brain ◽  
Zonula Occludens 1 ◽  
End Stage ◽  
Diffuse Midline Glioma

Abstract INTRODUCTION Chemotherapy has been unsuccessful for pediatric diffuse midline glioma (DMG) most likely due to an intact blood-brain barrier (BBB). However, the BBB has not been characterized in DMG and therefore its implications for drug delivery are unknown. In this study we characterized the BBB in DMG patients and compared this to healthy controls. METHODS End-stage DMG pontine samples (n=5) were obtained from the VUmc diffuse intrinsic pontine glioma (DIPG) autopsy study and age-matched healthy pontine samples (n=22) were obtained from the NIH NeuroBioBank. Tissues were stained for BBB markers claudin-5, zonula occludens-1, laminin, and PDGFRβ. Claudin-5 stains were used to determine vascular density and diameter. RESULTS In DMG, expression of claudin-5 was reduced and dislocated to the abluminal side of endothelial cells. In addition, the expression of zonula occludens-1 was reduced. The basement membrane protein laminin expression was reduced at the glia limitans in both pre-existent vessels and neovascular proliferation. PDGFRβ expression was not observed in DMG but was present in healthy pons. Furthermore, the number of blood vessels in DMG was significantly (P&lt; 0.01) reduced (13.9 ± 11.8/mm2) compared to healthy pons (26.3 ± 14.2/mm2). Markedly, the number of small blood vessels (&lt; 10µm) was significantly lower (P&lt; 0.01) while larger blood vessels (&gt; 10µm) were not significantly different (P= 0.223). The mean vascular diameter was larger for DMG 9.3 ± 9.9µm compared to 7.7 ± 9.0µm for healthy pons (P= 0.016). CONCLUSION Both the BBB and the vasculature are altered at end-stage DMG. The reduced vascular density might have implications for several drug delivery methods such as focused ultrasound and convection enhanced delivery that are being explored for the treatment of DMG. The functional effects of the structurally altered BBB remain unknown and further research is needed to evaluate the BBB integrity at end-stage DMG

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