Novel Pannexin-1-Coupled Signaling Cascade Involved in the Control of Endothelial Cell Function and NO-Dependent Relaxation
Journal
Oxidative Medicine and Cellular Longevity
ISSN
1942-0994
1942-0900
Date Issued
2021
Author(s)
Mauricio A. Lillo
Pablo S. Gaete
Pía C. Burboa
Inés Poblete
Xavier F. Figueroa
Type
Resource Types::text::journal::journal article
Abstract
<jats:p>Deletion of pannexin-1 (Panx-1) leads not only to a reduction in endothelium-derived hyperpolarization but also to an increase in NO-mediated vasodilation. Therefore, we evaluated the participation of Panx-1-formed channels in the control of membrane potential and [Ca2+]i of endothelial cells. Changes in NO-mediated vasodilation, membrane potential, superoxide anion (O2⋅–) formation, and endothelial cell [Ca2+]i were analyzed in rat isolated mesenteric arterial beds and primary cultures of mesenteric endothelial cells. Inhibition of Panx-1 channels with probenecid (1 mM) or the Panx-1 blocking peptide 10Panx (60 μM) evoked an increase in the ACh (100 nM)-induced vasodilation of KCl-contracted mesenteries and in the phosphorylation level of endothelial NO synthase (eNOS) at serine 1177 (P-eNOSS1177) and Akt at serine 473 (P-AktS473). In addition, probenecid or 10Panx application activated a rapid, tetrodotoxin (TTX, 300 nM)-sensitive, membrane potential depolarization and [Ca2+]i increase in endothelial cells. Interestingly, the endothelial cell depolarization was converted into a transient spike after removing Ca2+ ions from the buffer solution and in the presence of 100 μM mibefradil or 10 μM Ni2+. As expected, Ni2+ also abolished the increment in [Ca2+]i. Expression of Nav1.2, Nav1.6, and Cav3.2 isoforms of voltage-dependent Na+ and Ca2+ channels was confirmed by immunocytochemistry. Furthermore, the Panx-1 channel blockade was associated with an increase in O2⋅– production. Treatment with 10 μM TEMPOL or 100 μM apocynin prevented the increase in O2⋅– formation, ACh-induced vasodilation, P-eNOSS1177, and P-AktS473 observed in response to Panx-1 inhibition. These findings indicate that the Panx-1 channel blockade triggers a novel complex signaling pathway initiated by the sequential activation of TTX-sensitive Nav channels and Cav3.2 channels, leading to an increase in NO-mediated vasodilation through a NADPH oxidase-dependent P-eNOSS1177, which suggests that Panx-1 may be involved in the endothelium-dependent control of arterial blood pressure.</jats:p>
Cite this document
Lillo, M. A., Gaete, P. S., Puebla, M., Burboa, P. C., Poblete, I., & Figueroa, X. F. (2021). Novel pannexin‐1‐coupled signaling cascade involved in the control of endothelial cell function and no‐dependent relaxation. Oxidative Medicine and Cellular Longevity, 2021(1), 2678134. https://doi.org/10.1155/2021/2678134
Subjects
nadph oxidase activation
;
nitric-oxide
;
sodium-channels
;
k+-channels
;
depolarization
;
conductance
;
ca2+
;
phosphorylation
;
mechanisms
;
generation
;
amino acids
;
cells
;
control systems
;
cytology
;
depolarization
;
increments
;
inhibition
;
potassium compounds
;
animals
;
arteries
;
calcium channels
;
calcium signaling
;
connexins
;
endothelial cells
;
male
;
membrane potentials
;
nadph oxidases
;
nerve tissue proteins
;
nitric oxide
;
nitric oxide synthase type iii
;
phosphorylation
;
proto-oncogene proteins c-akt
;
rats, sprague-dawley
;
signal transduction
;
subcellular fractions
;
superoxides
;
tetrodotoxin
;
vascular resistance
;
vasodilation
;
amino acids
;
blood pressure
;
cytology
;
depolarization
;
endothelial cells
;
potassium compounds
;
apocynin
;
calcium
;
calcium channel
;
endothelial nitric oxide synthase
;
nitric oxide
;
oxygen
;
pannexin 1
;
potassium chloride
;
probenecid
;
protein
;
reduced nicotinamide adenine dinucleotide phosphate oxidase
;
sodium channel
;
superoxide
;
tempol
;
unclassified drug
;
calcium channel
;
endothelial nitric oxide synthase
;
gap junction protein
;
nerve protein
;
nitric oxide
;
pannexin 1, rat
;
protein kinase b
;
reduced nicotinamide adenine dinucleotide phosphate oxidase
;
superoxide
;
tetrodotoxin
;
arterial blood pressure
;
cell depolarization
;
immunocytochemistry
;
membrane potentials
;
sequential activation
;
signaling cascades
;
signaling pathways
;
superoxide anions
;
animal cell
;
animal experiment
;
animal tissue
;
article
;
cell function
;
controlled study
;
depolarization
;
endothelium cell
;
enzyme phosphorylation
;
immunocytochemistry
;
male
;
membrane depolarization
;
membrane potential
;
mesenteric arterial bed
;
mesentery
;
nonhuman
;
primary cell culture
;
protein expression
;
rat
;
signal transduction
;
vasodilatation
;
vasomotor reflex
;
animal
;
artery
;
calcium signaling
;
cell fractionation
;
drug effect
;
endothelium cell
;
metabolism
;
phosphorylation
;
sprague dawley rat
;
vascular resistance
;
cell signaling