Journal: Developmental biology

Article Title: The glycocalyx is present as soon as blood flow is initiated and is required for normal vascular development.

doi: 10.1016/j.ydbio.2012.07.009

Figure Lengend Snippet: Fig. 5. Vessel branching is altered by selective degradation of the glycocalyx. Whole-mount embryos were stained with QH1 antibody following 16-hour exposure to bead-immobilized enzymes (A–E). Hyal treatment induced regions of hyperfused vessels and stunted the growth of the vitelline artery (arrow, B). Injection of bead- immobilized ChABC resulted in a decrease in vascular density near the vitelline artery (arrow, C). Injection of HepIII and NeurV did not inhibit vascular remodeling (D, E). Images were obtained by capturing 40–60 images of the extra-embryonic yolk sac on a confocal microscope and reassembled using Adobe Photoshop’s photomerge function. The beads used for protein immobilization are fluorescent and create a speckle pattern in the blood vessels. Scale bar 1 mm.

Article Snippet: Endothelial cell were labeled with QH1 (Developmental Studies Hybridoma Bank, 1:100).

Techniques: Staining, Injection, Microscopy

Journal: International journal of cardiology

Article Title: Enhanced external counterpulsation inhibits endothelial apoptosis via modulation of BIRC2 and Apaf-1 genes in porcine hypercholesterolemia.

doi: 10.1016/j.ijcard.2013.11.033

Figure Lengend Snippet: Fig. 2. Isolation and identification of vascular endothelial cells from the aortic endothelium with collagenase. Representative photomicrographs (×400) of isolated cells treated histocytochemically with CD31 antibody and incubated with DiI-Ac-LDL. Cells with cytomembrane and cytoplasmic staining of amber color indicate vascular endothelial cells (black arrow), and cells with red fluorescence were identified as viable vascular endothelial cells (white arrow), which consisted of the majority of total treated cells, indicating a successful en- dothelial cell isolation from aortic endothelium.

Article Snippet: Then mouse polyclonal CD31 antibody (at 1:100 dilution; Boster Biological Technology, Inc, Wuhan, China) was applied and incubated at 37 °C for 1 h, then stained with SABC, enhanced by DAB, dehydrated in a graded ethanol series, and followed by dimethylbenzene treatment to improve transparency, and finally mounted with neutral gum.

Techniques: Isolation, Incubation, Staining, Cell Isolation

Journal: Cardiovascular research

Article Title: Intermedin induces loss of coronary microvascular endothelial barrier via derangement of actin cytoskeleton: role of RhoA and Rac1.

doi: 10.1093/cvr/cvr213

Figure Lengend Snippet: Figure 1 Intermedin (IMD) activates the cAMP/protein kinase A (PKA) pathway. (A) Intermedin induces cAMP production in rat cor- onary microvascular endothelial cells (RCECs). The RCECs were treated with increasing concentrations of IMD for 30 min, and the cAMP concentrations were measured by a colorimetric method. (B) The RCECs were exposed to IMD (10 nM) for the indicated time periods. n ¼ 3; *P , 0.05 vs. control. (C) Effect of IMD, aCGRP8–37 and PKI on VASP phosphorylation at Ser157. Represen- tative western blots of VASP phosphorylation. The RCECs were exposed to IMD (10 nM), aCGRP8–37 (1 mM), aCGRP8–37 plus IMD, PKI (20 mM), and PKI plus IMD or vehicle (C; control) for 30 min. The cells were incubated with PKI for 60 min or aCGRP8–37 for 30 min before IMD was added. The western blots are representative of three separate experiments with independent cell preparations.

Article Snippet: Rat aortic endothelial cells were purchased from CELL applications Inc. (San Diego, CA, USA) in passage 1 and cultured according to the instructions of the supplier.

Techniques: Control, Phospho-proteomics, Western Blot, Incubation

Journal: Cardiovascular research

Article Title: Intermedin induces loss of coronary microvascular endothelial barrier via derangement of actin cytoskeleton: role of RhoA and Rac1.

doi: 10.1093/cvr/cvr213

Figure Lengend Snippet: Figure 2 Effect of IMD on RCECs permeability. (A) RCEC monolayers were treated with IMD (0.1, 1, and 10 nM), FSK (5 mM), or vehicle (control) as indicated. Data are means +SD of five separate experiments with independent cell preparations. *P , 0.05 vs. control. (B) Concentration– response curve of the effect of IMD as in A on permeability after 30 min. The RCEC monolayers were exposed to IMD as in A or vehicle (control) as indicated. n ¼ 5; *P , 0.05 vs. control. (C) Effect of CGRP receptor inhibition on the IMD-induced increase in permeability. The RCEC monolayers were exposed to IMD (10 nM), aCGRP8–37 (1 mM) plus IMD, or vehicle (control) as indicated. n ¼ 3; *P , 0.05 vs. control; #P , 0.05 vs. IMD alone. (D) Concentration–response curve of the effect of aCGRP8–37 on IMD-induced hyperpermeability. The RCEC monolayers were exposed to IMD (10 nM), in the absence or presence of increasing concentrations of aCGRP8–37 or vehicle (control) as indicated. n ¼ 3; *P , 0.05 vs. IMD alone. (E) Effect of PKA inhibition on IMD-induced hyperpermeability. The RCEC monolayers were treated with IMD (10 nM), PKI (40 mM) plus IMD, or vehicle (control), as indicated. n ¼ 3; *P , 0.05 vs. control; #P , 0.05 vs. IMD alone. (F) Effect of cAMP analogues on RCEC monolayer permeability. The RCEC monolayers were treated with the PKA activator, 6-Bnz-cAMP (50 mM), the Epac activator, 8-CPT-cAMP (200 mM), or vehicle (control) as indicated. n ¼ 3; *P , 0.05 vs. control. (G) human umbilical vein endothelial cell (HUVEC) monolayers were exposed to IMD (10 nM), hAM22–52 (1 mM; a specific AM-receptor antagonist) plus IMD, FSK (5 mM), or vehicle (control) as indicated. n ¼ 3; *P , 0.05 vs. control; #P , 0.05 vs. IMD alone. (H ) The HUVEC monolayers were treated with IMD (10 nM), PKI (20 mM), PKI plus IMD, or vehicle (control; C), as indicated. n ¼ 3; *P , 0.05 vs. control; #P , 0.05 vs. IMD alone.

Article Snippet: Rat aortic endothelial cells were purchased from CELL applications Inc. (San Diego, CA, USA) in passage 1 and cultured according to the instructions of the supplier.

Techniques: Permeability, Control, Concentration Assay, Inhibition, Analogues

Journal: Brain : a journal of neurology

Article Title: Connexin43 mimetic peptide reduces vascular leak and retinal ganglion cell death following retinal ischaemia.

doi: 10.1093/brain/awr338

Figure Lengend Snippet: Figure 3 Three hours of hypoxia and 6 h of reperfusion lead to significant endothelial cell death in vitro in rat brain microvascular endothelial cells (RBMVEC; A). Non-specific gap junction blocker carbenoxolone, non-specific hemichannel blocker LaCl3, and connexin43 (Cx43) mimetic peptide protected endothelial cells against hypoxic injury, with the number of viable cells significantly higher than no treatment, while scrambled peptide did not have any protective effects. The number of viable cells was expressed as percentage of the control without hypoxia. Hypoxia and reperfusion also lead to significant propidium iodide dye uptake into primary rat brain microvascular endothelial cells indicating open hemichannels (B). Carbenoxolone, LaCl3 and connexin43 mimetic peptide significantly prevented dye uptake compared to no treatment, indicating hemichannel closure, while scrambled peptide did not have any effect. Stars denote statistical significance when compared to the control group or compared between groups in brackets; P 5 0.05.

Article Snippet: Rat brain microvascular endothelial cells (R840K-05a, Cell Applications) were plated into 24-well plates (1 105 cells/well) in rat brain microvascular endothelial cell growth media (R819K-500, Cell Applications) and allowed to settle for 16 h. Medium was then removed and replaced with Dulbecco’s Modified Eagle’s Medium/F12 containing 0.5% foetal bovine serum and 1% glutamine.

Techniques: In Vitro, Control