Individual Gap Junction Plaques Contain Multiple Connexins in Arterial Endothelium

Yeh, Hung‐I; Rothery, Stephen; Dupont, Emmanuel; Coppen, Steven R.; Severs, Nicholas J.

doi:10.1161/01.res.83.12.1248

Cited by 163 publications

(161 citation statements)

References 58 publications

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“…This approach demonstrates that neither antibody cross-reacts with other connexins. 13 More comprehensive characterization was performed by using immunogold labeling and electron microscopy. 13 For Cx43, a mouse monoclonal antibody of widely established reliability was purchased from Chemicon.…”

Section: Anti-connexin Antibodies and Endothelial Cell Markermentioning

confidence: 99%

Multiple Connexin Expression in Regenerating Arterial Endothelial Gap Junctions

Yeh

Lai²,

Chang³

et al. 2000

ATVB

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Abstract-Endothelial cells form gap junctions that, according to vessel type, may be composed of up to 3 types of connexin, connexin37, connexin40, and connexin43. Although changes in connexin expression have been linked to growth and injury in cultured endothelial cells, information on connexin expression in regenerating endothelium in situ is lacking. We investigated gap junction distribution and expression of all 3 endothelial connexins during healing in rat carotid artery after denudation injury. En face viewing of the vascular luminal surface by means of immunoconfocal microscopy was used to examine the spatial and temporal expression pattern of the endothelial connexins. Gap junction spots labeled by specific antisera against connexin37, connexin40, and connexin43 were quantified 7, 14, and 28 days after injury, and the relations among the connexins were examined by using colocalization analysis. Complementary electron microscopy was also conducted. After injury, the regenerating endothelium initially expressed small, sparse gap junctions, the numbers of which progressively increased to values equivalent to those of controls. Although connexin40 gap-junctional spot size and area returned to uninjured levels by 28 days after injury, connexin37 and connexin43 spot size and area exceeded those of the uninjured artery (PϽ0.05). Double-label analysis showed that even though colocalization of connexins to the same gap-junctional spot is a common feature, the extent of colocalization was time dependent (Ͼ80% in the intact artery at postinjury day 28 and Ͻ70% at postinjury days 7 and 14, PϽ0.01). We conclude that distinct alterations in expression of the 3 connexins are associated with regeneration of the arterial endothelium in situ, implying different intercellular communication requirements during the various phases of the healing process.

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Section: Anti-connexin Antibodies and Endothelial Cell Markermentioning

confidence: 99%

Multiple Connexin Expression in Regenerating Arterial Endothelial Gap Junctions

Yeh

Lai²,

Chang³

et al. 2000

ATVB

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“…The specificity of these antibodies has been demonstrated by western blotting and immunolabeling of HeLa cell transfectants, and by immunogold labeling (thin sections and/or freeze-fracture cytochemistry). 7,25 8 Macrophages were detected by a mouse anti-CD68 monoclonal antibody (1:50; Dako, catalog No. M0876), and T lymphocytes by a mouse anti-CD3 monoclonal antibody (1:200; Chemicon, catalog No.…”

Section: Immunofluorescence Labeling Of Gap-junctional Connexins Vasmentioning

confidence: 99%

Differential Expression of Connexin43 and Desmin Defines Two Subpopulations of Medial Smooth Muscle Cells in the Human Internal Mammary Artery

Yeh

Haw

et al. 1999

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Abstract-Upregulation of connexin43-gap junctions is associated with transition of contractile vascular smooth muscle cells (SMCs) to the synthetic state. To determine whether phenotypically distinct subpopulations of medial SMCs differentially express connexin43, we investigated the human distal internal mammary artery, a structurally heterogeneous vessel with features ranging from elastic to elastomuscular to muscular. Immunoconfocal microscopy combined with quantitative analysis and complemented by in situ hybridization showed that SMCs in the elastic medial regions expressed high levels of connexin43 but low levels of desmin, whereas those of muscular medial regions expressed low levels of connexin43 but high levels of desmin. Ultrastructurally, SMCs of both regions were of the contractile phenotype, but the former cells were irregular in shape with relatively prominent synthetic organelles whereas the latter were spindle shaped with fewer synthetic organelles. Vimentin, smooth muscle ␣-actin, calponin, h-caldesmon, and myosin heavy chains (SM1 and SM2) were equally highly expressed by most cells in both subpopulations. The connexin43/desmin expression pattern of SMCs in regions of intimal thickening resembled those of elastic medial regions. These findings refine the view suggested from previous studies that high levels of connexin43 expression are associated with SMCs of a less contractile/more synthetic phenotype. In the internal mammary artery, the 2 subpopulations of SMCs with markedly different connexin43 expression levels both represent a differentiated contractile phenotype, but the subpopulation showing high levels of connexin43-gap junctions is characterized by low levels of desmin and structural features that reflect a more synthetic tendency.

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“…Individually, these proteins form channels with different physiological properties (1)(2)(3). However, these connexins have all been found within the same cell type (the endothelial cell), and in a number of cases they can be identified within the same individual cell (4)(5)(6). The relative abundance of these connexins varies in different endothelia.…”

Section: Introductionmentioning

confidence: 99%

Modulation of intercellular communication by differential regulation and heteromeric mixing of co-expressed connexins

Beyer

Gemel

Seul

et al. 2000

Braz J Med Biol Res

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Intercellular communication may be regulated by the differential expression of subunit gap junction proteins (connexins) which form channels with differing gating and permeability properties. Endothelial cells express three different connexins (connexin37, connexin40, and connexin43) in vivo. To study the differential regulation of expression and synthesis of connexin37 and connexin43, we used cultured bovine aortic endothelial cells which contain these two connexins in vitro. RNA blots demonstrated discordant expression of these two connexins during growth to confluency. RNA blots and immunoblots showed that levels of these connexins were modulated by treatment of cultures with transforming growth factor-ß1. To examine the potential ability of these connexins to form heteromeric channels (containing different connexins within the same hemi-channel), we stably transfected connexin43-containing normal rat kidney (NRK) cells with connexin37 or connexin40. In the transfected cells, both connexin proteins were abundantly produced and localized in identical distributions as detected by immunofluorescence. Double whole-cell patch-clamp studies showed that co-expressing cells exhibited unitary channel conductances and gating characteristics that could not be explained by hemi-channels formed of either connexin alone. These observations suggest that these connexins can readily mix with connexin43 to form heteromeric channels and that the intercellular communication between cells is determined not only by the properties of individual connexins, but also by the interactions of those connexins to form heteromeric channels with novel properties. Furthermore, modulation of levels of the co-expressed connexins during cell proliferation or by cytokines may alter the relative abundance of different heteromeric combinations.

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Individual Gap Junction Plaques Contain Multiple Connexins in Arterial Endothelium

Cited by 163 publications

References 58 publications

Multiple Connexin Expression in Regenerating Arterial Endothelial Gap Junctions

Multiple Connexin Expression in Regenerating Arterial Endothelial Gap Junctions

Differential Expression of Connexin43 and Desmin Defines Two Subpopulations of Medial Smooth Muscle Cells in the Human Internal Mammary Artery

Modulation of intercellular communication by differential regulation and heteromeric mixing of co-expressed connexins

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