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Beta-Adrenoceptor Activation Reduces Both Dermal Microvascular Endothelial Cell Migration via a cAMP-Dependent Mechanism and Wound Angiogenesis.

O'Leary AP, Fox JM, Pullar CE - J. Cell. Physiol. (2015)

Bottom Line: They play a role in wound repair but their specific role in angiogenesis is unknown.In this study, a range of in vitro assays (single cell migration, scratch wound healing, ELISAs for angiogenic growth factors and tubule formation) were performed with human dermal microvascular endothelial cells (HDMEC) to investigate and dissect mechanisms underpinning β-AR-mediated modulation of angiogenesis in chick chorioallantoic membranes (CAM) and murine excisional skin wounds. β-AR activation reduced HDMEC migration via cyclic adenosine monophosphate (cAMP)-dependent and protein kinase A (PKA)-independent mechanisms as demonstrated through use of an EPAC agonist that auto-inhibited the cAMP-mediated β-AR transduced reduction in HDMEC motility; a PKA inhibitor was, conversely, ineffective.In more complex environments, β-AR activation delayed HDMEC tubule formation and decreased angiogenesis both in the CAM assay and in murine excisional skin wounds in vivo. β-AR activation reduced HDMEC function in vitro and angiogenesis in vivo; therefore, β-AR agonists could be promising anti-angiogenic modulators in skin.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Physiology and Pharmacology, University of Leicester, Leicester, UK.

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β-AR activation and sp cAMP decreases embryonic angiogenesis. CAM assays were performed as described. The CAMs were treated with 10 µM Iso or 50 µM sp cAMP at day 5. Eggs were imaged every 24 h until day 10. Images depicting representative angiogenesis 9 days post fertilisation are presented; scale bar = 1 mm (A). Images were analysed by counting the total number of vessel branch points per field of view. The mean total number of vessel branch points per field of view was then calculated for control and treatment groups to give a mean amount of angiogenesis. The data shown were combined from 3–16 independent experiments using a total of 28 eggs (control N = 16; Iso N = 9; sp cAMP N = 3). Data were statistically analysed, normalised to the control mean value for angiogenesis and graphically represented with the bars representing the means ± SEM. (** P < 0.01) (B).
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fig07: β-AR activation and sp cAMP decreases embryonic angiogenesis. CAM assays were performed as described. The CAMs were treated with 10 µM Iso or 50 µM sp cAMP at day 5. Eggs were imaged every 24 h until day 10. Images depicting representative angiogenesis 9 days post fertilisation are presented; scale bar = 1 mm (A). Images were analysed by counting the total number of vessel branch points per field of view. The mean total number of vessel branch points per field of view was then calculated for control and treatment groups to give a mean amount of angiogenesis. The data shown were combined from 3–16 independent experiments using a total of 28 eggs (control N = 16; Iso N = 9; sp cAMP N = 3). Data were statistically analysed, normalised to the control mean value for angiogenesis and graphically represented with the bars representing the means ± SEM. (** P < 0.01) (B).

Mentions: To investigate the effect of β-AR activation on embryonic angiogenesis, the CAM assay was performed, as described (Ausprunk et al., 1974). Representative images of CAMs 9 days post-fertilisation are presented (Fig. 7A). Iso significantly decreased angiogenesis by 45%. Similarly, an active cAMP analogue, sp cAMP, significantly decreased angiogenesis by 51% (Fig. 7B).


Beta-Adrenoceptor Activation Reduces Both Dermal Microvascular Endothelial Cell Migration via a cAMP-Dependent Mechanism and Wound Angiogenesis.

O'Leary AP, Fox JM, Pullar CE - J. Cell. Physiol. (2015)

β-AR activation and sp cAMP decreases embryonic angiogenesis. CAM assays were performed as described. The CAMs were treated with 10 µM Iso or 50 µM sp cAMP at day 5. Eggs were imaged every 24 h until day 10. Images depicting representative angiogenesis 9 days post fertilisation are presented; scale bar = 1 mm (A). Images were analysed by counting the total number of vessel branch points per field of view. The mean total number of vessel branch points per field of view was then calculated for control and treatment groups to give a mean amount of angiogenesis. The data shown were combined from 3–16 independent experiments using a total of 28 eggs (control N = 16; Iso N = 9; sp cAMP N = 3). Data were statistically analysed, normalised to the control mean value for angiogenesis and graphically represented with the bars representing the means ± SEM. (** P < 0.01) (B).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263239&req=5

fig07: β-AR activation and sp cAMP decreases embryonic angiogenesis. CAM assays were performed as described. The CAMs were treated with 10 µM Iso or 50 µM sp cAMP at day 5. Eggs were imaged every 24 h until day 10. Images depicting representative angiogenesis 9 days post fertilisation are presented; scale bar = 1 mm (A). Images were analysed by counting the total number of vessel branch points per field of view. The mean total number of vessel branch points per field of view was then calculated for control and treatment groups to give a mean amount of angiogenesis. The data shown were combined from 3–16 independent experiments using a total of 28 eggs (control N = 16; Iso N = 9; sp cAMP N = 3). Data were statistically analysed, normalised to the control mean value for angiogenesis and graphically represented with the bars representing the means ± SEM. (** P < 0.01) (B).
Mentions: To investigate the effect of β-AR activation on embryonic angiogenesis, the CAM assay was performed, as described (Ausprunk et al., 1974). Representative images of CAMs 9 days post-fertilisation are presented (Fig. 7A). Iso significantly decreased angiogenesis by 45%. Similarly, an active cAMP analogue, sp cAMP, significantly decreased angiogenesis by 51% (Fig. 7B).

Bottom Line: They play a role in wound repair but their specific role in angiogenesis is unknown.In this study, a range of in vitro assays (single cell migration, scratch wound healing, ELISAs for angiogenic growth factors and tubule formation) were performed with human dermal microvascular endothelial cells (HDMEC) to investigate and dissect mechanisms underpinning β-AR-mediated modulation of angiogenesis in chick chorioallantoic membranes (CAM) and murine excisional skin wounds. β-AR activation reduced HDMEC migration via cyclic adenosine monophosphate (cAMP)-dependent and protein kinase A (PKA)-independent mechanisms as demonstrated through use of an EPAC agonist that auto-inhibited the cAMP-mediated β-AR transduced reduction in HDMEC motility; a PKA inhibitor was, conversely, ineffective.In more complex environments, β-AR activation delayed HDMEC tubule formation and decreased angiogenesis both in the CAM assay and in murine excisional skin wounds in vivo. β-AR activation reduced HDMEC function in vitro and angiogenesis in vivo; therefore, β-AR agonists could be promising anti-angiogenic modulators in skin.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Physiology and Pharmacology, University of Leicester, Leicester, UK.

Show MeSH
Related in: MedlinePlus