Connexin and Pannexin Large-Pore Channels in Microcirculation and Neurovascular Coupling Function

Burboa, Pía C.; Puebla, Mariela; Gaete, Pablo S.; Durán, Walter N.; Lillo, Mauricio A.

doi:10.3390/ijms23137303

Cited by 5 publications

(5 citation statements)

References 313 publications

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“…ECs are pivotal in maintaining vascular homeostasis through the regulation of permeability and vasoconstriction, both of which are tightly controlled by cytosolic Ca 2+ ([Ca 2+ ] c ) levels. In postcapillary veins, a [Ca 2+ ] c increase is crucial for promoting hyperpermeability [24–27], whereas in arterioles, [Ca 2+ ] c mediates the response to vasoactive agents such as nitric oxide (NO), endothelium hyperpolarizing factor, and endothelin‐1 [5]. Given the importance of Ca 2+ signaling in endothelial function, understanding how different matrix gels may influence Ca 2+ responses to endothelium agonists is of significant interest.…”

Section: Resultsmentioning

confidence: 99%

Impact of Matrix Gel Variations on Primary Culture of Microvascular Endothelial Cell Function

Burboa,

Corrêa‐Velloso,

Arriagada

et al. 2024

Microcirculation

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ObjectiveThe endothelium regulates crucial aspects of vascular function, including hemostasis, vasomotor tone, proliferation, immune cell adhesion, and microvascular permeability. Endothelial cells (ECs), especially in arterioles, are pivotal for flow distribution and peripheral resistance regulation. Investigating vascular endothelium physiology, particularly in microvascular ECs, demands precise isolation and culturing techniques.MethodsFreshly isolated ECs are vital for examining protein expression, ion channel behavior, and calcium dynamics. Establishing primary endothelial cell cultures is crucial for unraveling vascular functions and understanding intact microvessel endothelium roles. Despite the significance, detailed protocols and comparisons with intact vessels are scarce in microvascular research. We developed a reproducible method to isolate microvascular ECs, assessing substrate influence by cultivating cells on fibronectin and gelatin matrix gels. This comparative approach enhances our understanding of microvascular endothelial cell biology.ResultsMicrovascular mesenteric ECs expressed key markers (VE‐cadherin and eNOS) in both matrix gels, confirming cell culture purity. Under uncoated conditions, ECs were undetected, whereas proteins linked to smooth muscle cells and fibroblasts were evident.Examining endothelial cell (EC) physiological dynamics on distinct matrix substrates revealed comparable cell length, shape, and Ca2+ elevations in both male and female ECs on gelatin and fibronectin matrix gels. Gelatin‐cultured ECs exhibited analogous membrane potential responses to acetylcholine (ACh) or adenosine triphosphate (ATP), contrasting with their fibronectin‐cultured counterparts. In the absence of stimulation, fibronectin‐cultured ECs displayed a more depolarized resting membrane potential than gelatin‐cultured ECs.ConclusionsGelatin‐cultured ECs demonstrated electrical behaviors akin to intact endothelium from mouse mesenteric arteries, thus advancing our understanding of endothelial cell behavior within diverse microenvironments.

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Section: Resultsmentioning

confidence: 99%

Impact of Matrix Gel Variations on Primary Culture of Microvascular Endothelial Cell Function

Burboa,

Corrêa‐Velloso,

Arriagada

et al. 2024

Microcirculation

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“…11,63 As a result, Panx channels are associated with a wide range of cellular physiological and pathophysiological functions. 17,64 Studies demonstrated that Panxs participate in various biological processes, including inflammation, 8,65 ATP signaling, longrange Ca 2+ wave propagation, 66 synaptic plasticity regulation, 67 vascular homeostasis, 68 and neurotoxicity. 7 Additionally, Panxs may contribute to tumor suppression, ischemic cell death, atherosclerosis, apoptosis, 11,69 human immunodeficiency virus (HIV), and epileptic seizures.…”

Section: Panxs Formed Channelsmentioning

confidence: 99%

Pannexins in the musculoskeletal system: new targets for development and disease progression

Luo,

Zheng,

Xiao

et al. 2024

Bone Res

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During cell differentiation, growth, and development, cells can respond to extracellular stimuli through communication channels. Pannexin (Panx) family and connexin (Cx) family are two important types of channel-forming proteins. Panx family contains three members (Panx1-3) and is expressed widely in bone, cartilage and muscle. Although there is no sequence homology between Panx family and Cx family, they exhibit similar configurations and functions. Similar to Cxs, the key roles of Panxs in the maintenance of physiological functions of the musculoskeletal system and disease progression were gradually revealed later. Here, we seek to elucidate the structure of Panxs and their roles in regulating processes such as osteogenesis, chondrogenesis, and muscle growth. We also focus on the comparison between Cx and Panx. As a new key target, Panxs expression imbalance and dysfunction in muscle and the therapeutic potentials of Panxs in joint diseases are also discussed.

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“…TRPV4-activation induced a strongly ethidium uptake in ECs during the time (Figure 2D). Either inhibition of NO production with 100 µM L-NAME or the blockade of Cx43 hemichannels with 50 µM β-glycyrrhetinic acid (β-GA) or 50 µM Gap19-TAT, a specific Cx43 hemichannel blocker 15,16,24,25 , significantly reduced dye uptake increases. (Figure 2D).…”

Section: Trpv4 Associates Closely With Cx43 and Its Activation Promot...mentioning

confidence: 99%

“…ECs were incubated in MOPS-buffered physiological saline solution (pH 7.4) containing 5 μM ethidium bromide to assess hemichannel activity a well-established method to evaluate connexin hemichannel function 14,16,24,75,76 . The opening of Cx hemichannels was evaluated by tracking the uptake of ethidium over time.…”

Section: Hemichannel Activitymentioning

confidence: 99%

Endothelial TRPV4/Cx43 Signaling Complex Regulates Vasomotor Tone in Resistance Arteries

Burboa,

Gaete,

Shu

et al. 2024

Preprint

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S-nitrosylation of Cx43 gap junction channels critically regulates communication between smooth muscle cells and endothelial cells. This posttranslational modification also induces the opening of undocked Cx43 hemichannels. However, its specific impact on vasomotor regulation remains unclear. Considering the role of endothelial TRPV4 channel activation in promoting vasodilation through nitric oxide (NO) production, we investigated the direct modulation of endothelial Cx43 hemichannels by TRPV4 channel activation. Using the proximity ligation assay, we identify that Cx43 and TRPV4 are found in close proximity in the endothelium of resistance arteries. In primary endothelial cell cultures from resistance arteries (ECs), GSK-induced TRPV4 activation enhances eNOS activity, increases NO production, and opens Cx43 hemichannels via direct S-nitrosylation. Notably, the elevated intracellular Ca2+ levels caused by TRPV4 activation were reduced by blocking Cx43 hemichannels. In ex vivo mesenteric arteries, inhibiting Cx43 hemichannels reduced endothelial hyperpolarization without affecting NO production in ECs, underscoring a critical role of TRPV4/Cx43 signaling in endothelial electrical behavior. We perturbed the proximity of Cx43/TRPV4 by disrupting lipid rafts in ECs using β-cyclodextrin. Under these conditions, hemichannel activity, Ca2+ influx, and endothelial hyperpolarization were blunted upon GSK stimulation. Intravital microscopy of mesenteric arterioles in vivo further demonstrated that inhibiting Cx43 hemichannels activity, NO production and disrupting endothelial integrity reduce TRPV4-induced relaxation. These findings underscore a new pivotal role of Cx43 hemichannel associated with TRPV4 signaling pathway in modulating endothelial electrical behavior and vasomotor tone regulation.

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Connexin and Pannexin Large-Pore Channels in Microcirculation and Neurovascular Coupling Function

Cited by 5 publications

References 313 publications

Impact of Matrix Gel Variations on Primary Culture of Microvascular Endothelial Cell Function

Impact of Matrix Gel Variations on Primary Culture of Microvascular Endothelial Cell Function

Pannexins in the musculoskeletal system: new targets for development and disease progression

Endothelial TRPV4/Cx43 Signaling Complex Regulates Vasomotor Tone in Resistance Arteries

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