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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 delays the formation of tubule-like structures in HDMECs. Tubule assays were performed as described in the methods. Untreated HDMECs or those treated with 10 μM Iso were imaged at 0 and 6 h. Images representative of control and Iso-treated cells at 6 h are presented; scale bar = 100 μm (A). Tubule formation was analysed by counting the number of tubule-like structures in five images, taken at pre-determined positions in each well, in duplicate. The data shown were combined from five independent experiments, from three separate cell strains. Data were averaged, statistically analysed and graphically represented with the bars representing the means ± SEM. (* P < 0.05) (B).
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fig06: β-AR activation delays the formation of tubule-like structures in HDMECs. Tubule assays were performed as described in the methods. Untreated HDMECs or those treated with 10 μM Iso were imaged at 0 and 6 h. Images representative of control and Iso-treated cells at 6 h are presented; scale bar = 100 μm (A). Tubule formation was analysed by counting the number of tubule-like structures in five images, taken at pre-determined positions in each well, in duplicate. The data shown were combined from five independent experiments, from three separate cell strains. Data were averaged, statistically analysed and graphically represented with the bars representing the means ± SEM. (* P < 0.05) (B).

Mentions: ECs undergo numerous physiological processes that contribute to angiogenesis, these include: invasion, alignment, elongation and apoptosis in addition to migration and proliferation (Bauer et al., 2005; Eming et al., 2007). To explore HDMEC physiological processes in a more complex environment, HDMECs were cultured on top of BME for 24 h in the presence or absence of Iso. HDMECs formed tubule-like structures after 6 h (Figs. 6A and B). To quantitate HDMEC tubule development, the number of tubule-like structures were counted. After 6 h, β-AR activation significantly delayed the formation of tubule-like structures by 14% (Fig. 6B).


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 delays the formation of tubule-like structures in HDMECs. Tubule assays were performed as described in the methods. Untreated HDMECs or those treated with 10 μM Iso were imaged at 0 and 6 h. Images representative of control and Iso-treated cells at 6 h are presented; scale bar = 100 μm (A). Tubule formation was analysed by counting the number of tubule-like structures in five images, taken at pre-determined positions in each well, in duplicate. The data shown were combined from five independent experiments, from three separate cell strains. Data were averaged, statistically analysed and graphically represented with the bars representing the means ± SEM. (* P < 0.05) (B).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig06: β-AR activation delays the formation of tubule-like structures in HDMECs. Tubule assays were performed as described in the methods. Untreated HDMECs or those treated with 10 μM Iso were imaged at 0 and 6 h. Images representative of control and Iso-treated cells at 6 h are presented; scale bar = 100 μm (A). Tubule formation was analysed by counting the number of tubule-like structures in five images, taken at pre-determined positions in each well, in duplicate. The data shown were combined from five independent experiments, from three separate cell strains. Data were averaged, statistically analysed and graphically represented with the bars representing the means ± SEM. (* P < 0.05) (B).
Mentions: ECs undergo numerous physiological processes that contribute to angiogenesis, these include: invasion, alignment, elongation and apoptosis in addition to migration and proliferation (Bauer et al., 2005; Eming et al., 2007). To explore HDMEC physiological processes in a more complex environment, HDMECs were cultured on top of BME for 24 h in the presence or absence of Iso. HDMECs formed tubule-like structures after 6 h (Figs. 6A and B). To quantitate HDMEC tubule development, the number of tubule-like structures were counted. After 6 h, β-AR activation significantly delayed the formation of tubule-like structures by 14% (Fig. 6B).

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