Limits...
ADAM12 and ADAM17 are essential molecules for hypoxia-induced impairment of neural vascular barrier function.

Cui D, Arima M, Takubo K, Kimura T, Horiuchi K, Minagawa T, Matsuda S, Ikeda E - Sci Rep (2015)

Bottom Line: Therefore, the molecules involved in hypoxia-induced impairment of vascular barrier can be the targets to establish new therapies for intractable diseases.Hypoxic disappearance of claudin-5 from cell membranes and the consequent loss of barrier properties were completely suppressed by inhibition of the metalloproteinase activity which was found to be attributed to ADAM12 and ADAM17.This is the first report to specify the molecules which are responsible for hypoxia-induced impairment of neural vascular barrier and furthermore can be the targets of new therapeutic strategies for intractable neural diseases.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505, Japan.

ABSTRACT
Neural vascular barrier is essential for the life of multicellular organisms, and its impairment by tissue hypoxia is known to be a central of pathophysiology accelerating the progression of various intractable neural diseases. Therefore, the molecules involved in hypoxia-induced impairment of vascular barrier can be the targets to establish new therapies for intractable diseases. Here, we demonstrate that a disintegrin and metalloproteinases (ADAMs) 12 and 17 expressed in endothelial cells are the molecules responsible for the impairment of neural vascular barrier by hypoxia. Brain microvascular endothelial cells in vitro lost their barrier properties immediately after hypoxic stimulation through diminished localization of claudin-5, a tight junction molecule, on cell membranes. Hypoxic disappearance of claudin-5 from cell membranes and the consequent loss of barrier properties were completely suppressed by inhibition of the metalloproteinase activity which was found to be attributed to ADAM12 and ADAM17. Inhibition of either ADAM12 or ADAM17 was sufficient to rescue the in vivo neural vasculature under hypoxia from the loss of barrier function. This is the first report to specify the molecules which are responsible for hypoxia-induced impairment of neural vascular barrier and furthermore can be the targets of new therapeutic strategies for intractable neural diseases.

No MeSH data available.


Related in: MedlinePlus

ADAM12 as well as ADAM17 are essential for the hypoxia-induced disappearance of claudin-5 from endothelial cell membranes.(a and b) Immunofluorescence images (a) and their corresponding quantitative analysis of claudin-5 levels on cell membranes (b) in bEnd.3 monolayers treated with CHX, MG-132, non-silencing siRNA (NC siRNA, for negative control), siRNA for ADAM12 and siRNA for ADAM17 as described. With CHX-treated cells, it is clearly shown that the suppression of either ADAM12 or ADAM17 expression with siRNA rescues the cells from the hypoxia-accelerated decrease in claudin-5 levels on cell membranes, while it has no significant effects on the oxygen concentration-independent decrease. Oxygen concentration-independent decrease in claudin-5 is abolished by inhibition of ubiquitin-proteasome system with MG-132. NC siRNA; non-silencing siRNA for negative control. *P < 0.05; ns, not significant.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4525292&req=5

f3: ADAM12 as well as ADAM17 are essential for the hypoxia-induced disappearance of claudin-5 from endothelial cell membranes.(a and b) Immunofluorescence images (a) and their corresponding quantitative analysis of claudin-5 levels on cell membranes (b) in bEnd.3 monolayers treated with CHX, MG-132, non-silencing siRNA (NC siRNA, for negative control), siRNA for ADAM12 and siRNA for ADAM17 as described. With CHX-treated cells, it is clearly shown that the suppression of either ADAM12 or ADAM17 expression with siRNA rescues the cells from the hypoxia-accelerated decrease in claudin-5 levels on cell membranes, while it has no significant effects on the oxygen concentration-independent decrease. Oxygen concentration-independent decrease in claudin-5 is abolished by inhibition of ubiquitin-proteasome system with MG-132. NC siRNA; non-silencing siRNA for negative control. *P < 0.05; ns, not significant.

Mentions: In CHX-treated cells, the suppression of either ADAM12 or ADAM17 expression rescued the endothelial cells from hypoxic disappearance of claudin-5 to the level of CHX-treated normoxic cells, and the additional blockade of ubiquitin-proteasome with MG-132 led to the further increase in the claudin-5 immunofluorescence intensity to the level of normoxic cells without CHX treatment (Fig. 3a,b). These results clearly indicate the indispensable role of ADAM12 as well as ADAM17 in the hypoxia-induced disappearance of claudin-5 from cell membranes and consequently the impairment of neural vascular barrier function under hypoxia.


ADAM12 and ADAM17 are essential molecules for hypoxia-induced impairment of neural vascular barrier function.

Cui D, Arima M, Takubo K, Kimura T, Horiuchi K, Minagawa T, Matsuda S, Ikeda E - Sci Rep (2015)

ADAM12 as well as ADAM17 are essential for the hypoxia-induced disappearance of claudin-5 from endothelial cell membranes.(a and b) Immunofluorescence images (a) and their corresponding quantitative analysis of claudin-5 levels on cell membranes (b) in bEnd.3 monolayers treated with CHX, MG-132, non-silencing siRNA (NC siRNA, for negative control), siRNA for ADAM12 and siRNA for ADAM17 as described. With CHX-treated cells, it is clearly shown that the suppression of either ADAM12 or ADAM17 expression with siRNA rescues the cells from the hypoxia-accelerated decrease in claudin-5 levels on cell membranes, while it has no significant effects on the oxygen concentration-independent decrease. Oxygen concentration-independent decrease in claudin-5 is abolished by inhibition of ubiquitin-proteasome system with MG-132. NC siRNA; non-silencing siRNA for negative control. *P < 0.05; ns, not significant.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4525292&req=5

f3: ADAM12 as well as ADAM17 are essential for the hypoxia-induced disappearance of claudin-5 from endothelial cell membranes.(a and b) Immunofluorescence images (a) and their corresponding quantitative analysis of claudin-5 levels on cell membranes (b) in bEnd.3 monolayers treated with CHX, MG-132, non-silencing siRNA (NC siRNA, for negative control), siRNA for ADAM12 and siRNA for ADAM17 as described. With CHX-treated cells, it is clearly shown that the suppression of either ADAM12 or ADAM17 expression with siRNA rescues the cells from the hypoxia-accelerated decrease in claudin-5 levels on cell membranes, while it has no significant effects on the oxygen concentration-independent decrease. Oxygen concentration-independent decrease in claudin-5 is abolished by inhibition of ubiquitin-proteasome system with MG-132. NC siRNA; non-silencing siRNA for negative control. *P < 0.05; ns, not significant.
Mentions: In CHX-treated cells, the suppression of either ADAM12 or ADAM17 expression rescued the endothelial cells from hypoxic disappearance of claudin-5 to the level of CHX-treated normoxic cells, and the additional blockade of ubiquitin-proteasome with MG-132 led to the further increase in the claudin-5 immunofluorescence intensity to the level of normoxic cells without CHX treatment (Fig. 3a,b). These results clearly indicate the indispensable role of ADAM12 as well as ADAM17 in the hypoxia-induced disappearance of claudin-5 from cell membranes and consequently the impairment of neural vascular barrier function under hypoxia.

Bottom Line: Therefore, the molecules involved in hypoxia-induced impairment of vascular barrier can be the targets to establish new therapies for intractable diseases.Hypoxic disappearance of claudin-5 from cell membranes and the consequent loss of barrier properties were completely suppressed by inhibition of the metalloproteinase activity which was found to be attributed to ADAM12 and ADAM17.This is the first report to specify the molecules which are responsible for hypoxia-induced impairment of neural vascular barrier and furthermore can be the targets of new therapeutic strategies for intractable neural diseases.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505, Japan.

ABSTRACT
Neural vascular barrier is essential for the life of multicellular organisms, and its impairment by tissue hypoxia is known to be a central of pathophysiology accelerating the progression of various intractable neural diseases. Therefore, the molecules involved in hypoxia-induced impairment of vascular barrier can be the targets to establish new therapies for intractable diseases. Here, we demonstrate that a disintegrin and metalloproteinases (ADAMs) 12 and 17 expressed in endothelial cells are the molecules responsible for the impairment of neural vascular barrier by hypoxia. Brain microvascular endothelial cells in vitro lost their barrier properties immediately after hypoxic stimulation through diminished localization of claudin-5, a tight junction molecule, on cell membranes. Hypoxic disappearance of claudin-5 from cell membranes and the consequent loss of barrier properties were completely suppressed by inhibition of the metalloproteinase activity which was found to be attributed to ADAM12 and ADAM17. Inhibition of either ADAM12 or ADAM17 was sufficient to rescue the in vivo neural vasculature under hypoxia from the loss of barrier function. This is the first report to specify the molecules which are responsible for hypoxia-induced impairment of neural vascular barrier and furthermore can be the targets of new therapeutic strategies for intractable neural diseases.

No MeSH data available.


Related in: MedlinePlus