Multicolor and Electron Microscopic Imaging of Connexin Trafficking

Gaietta, Guido; Deerinck, Thomas J.; Adams, Stephen; Bouwer, James C.; Tour, Oded; Laird, Dale W.; Sosinsky, Gina E.; Tsien, Roger Y.; Ellisman, Mark H.

doi:10.1126/science.1068793

Cited by 865 publications

(706 citation statements)

References 26 publications

Supporting

Mentioning

671

Contrasting

Unclassified

Order By: Relevance

“…More recent studies using epitope and fluorescent protein-tagged Cx43 support the mechanism of GJ internalization by annular junctions Gaietta et al, 2002]. Mechanistic analyses revealed that components of the clathrin-dependent endocytosis machinery are directly involved in the internalization, inward movement, and initial degradation of these intercellular double-membrane vesicles.…”

Section: Gap Junction Removal and Degradationmentioning

confidence: 94%

“…Connexon-containing cargo-vesicles may fuse with the plasma membrane in the center of plaques or at their marginal boundaries, followed by lateral diffusion in the plasma membrane to reach the plaque site. According to Gaietta et al [2002], the latter pathway seems to be the preferred route for plaque renewal with removal of older connexons from the center of the plaques. Incorporation of new connexons preferentially at the periphery of plaques indicated that a directed transport of connexons via cargo-vesicles is fundamental for GJ assembly (see also below).…”

Section: Shuttling To the Plasma Membrane And Intramembrane Movementmentioning

confidence: 99%

See 1 more Smart Citation

Connexins, cell motility, and the cytoskeleton

Olk

Zoidl

Dermietzel

2009

Cell Motil. Cytoskeleton

View full text Add to dashboard Cite

Connexins (Cx) comprise a family of transmembrane proteins, which form intercellular channels between plasma membranes of two adjoining cells, commonly known as gap junctions. Recent reports revealed that Cx proteins interact with diverse cellular components to form a multiprotein complex, which has been termed “Nexus”. Potential interaction partners include proteins such as cytoskeletal proteins, scaffolding proteins, protein kinases and phosphatases. These interactions allow correct subcellular localization of Cxs and functional regulation of gap junction‐mediated intercellular communication. Evidence is accruing that Cxs might have channel‐independent functions, which potentially include regulation of cell migration, cell polarization and growth control. In the current review, we summarize recent knowledge on Cx interactions with cytoskeletal proteins and highlight some aspects of their role in cellular motility. Cell Motil. Cytoskeleton 66: 1000–1016, 2009. © 2009 Wiley‐Liss, Inc.

show abstract

Section: Gap Junction Removal and Degradationmentioning

confidence: 94%

Section: Shuttling To the Plasma Membrane And Intramembrane Movementmentioning

confidence: 99%

Connexins, cell motility, and the cytoskeleton

Olk

Zoidl

Dermietzel

2009

Cell Motil. Cytoskeleton

View full text Add to dashboard Cite

show abstract

“…While hemichannels are inevitably present in gap junction expressing cells because gap junctions are formed after docking to newly assembled hemichannels in adjacent cells [66], their opening, which becomes prominent under pathological conditions, could result in release of a series of biologically relevant signaling molecules such as ATP, glutamate, glutathione, NAD + , and prostaglandin E2 [65]. Hemichannels made by Cx43 or structurally related connexins, if presented in mature ORNs, will be impaired in OlfDNCX.…”

Section: Discussionmentioning

confidence: 99%

Gap junctions in olfactory neurons modulate olfactory sensitivity

Zhang

2010

BMC Neurosci

View full text Add to dashboard Cite

BackgroundOne of the fundamental questions in olfaction is whether olfactory receptor neurons (ORNs) behave as independent entities within the olfactory epithelium. On the basis that mature ORNs express multiple connexins, I postulated that gap junctional communication modulates olfactory responses in the periphery and that disruption of gap junctions in ORNs reduces olfactory sensitivity. The data collected from characterizing connexin 43 (Cx43) dominant negative transgenic mice OlfDNCX, and from calcium imaging of wild type mice (WT) support my hypothesis.ResultsI generated OlfDNCX mice that express a dominant negative Cx43 protein, Cx43/β-gal, in mature ORNs to inactivate gap junctions and hemichannels composed of Cx43 or other structurally related connexins. Characterization of OlfDNCX revealed that Cx43/β-gal was exclusively expressed in areas where mature ORNs resided. Real time quantitative PCR indicated that cellular machineries of OlfDNCX were normal in comparison to WT. Electroolfactogram recordings showed decreased olfactory responses to octaldehyde, heptaldehyde and acetyl acetate in OlfDNCX compared to WT. Octaldehyde-elicited glomerular activity in the olfactory bulb, measured according to odor-elicited c-fos mRNA upregulation in juxtaglomerular cells, was confined to smaller areas of the glomerular layer in OlfDNCX compared to WT. In WT mice, octaldehyde sensitive neurons exhibited reduced response magnitudes after application of gap junction uncoupling reagents and the effects were specific to subsets of neurons.ConclusionsMy study has demonstrated that altered assembly of Cx43 or structurally related connexins in ORNs modulates olfactory responses and changes olfactory activation maps in the olfactory bulb. Furthermore, pharmacologically uncoupling of gap junctions reduces olfactory activity in subsets of ORNs. These data suggest that gap junctional communication or hemichannel activity plays a critical role in maintaining olfactory sensitivity and odor perception.

show abstract

“…A small peptide with a bright associated dye would be an ideal tag and may be possible. The FlAsH/ReAsH peptide tagging system [110, 111] is currently the smallest commercially available tag. One of the major shortcomings is the lack of suitability for the secretory pathway.…”

Section: Perspectivementioning

confidence: 99%

Approaches to imaging unfolded secretory protein stress in living cells

Lajoie

Fazio

Snapp

2014

Endoplasmic Reticulum Stress in Diseases

View full text Add to dashboard Cite

The endoplasmic reticulum (ER) is the point of entry of proteins into the secretory pathway. Nascent peptides interact with the ER quality control machinery that ensures correct folding of the nascent proteins. Failure to properly fold proteins can lead to loss of protein function and cytotoxic aggregation of misfolded proteins that can lead to cell death. To cope with increases in the ER unfolded secretory protein burden, cells have evolved the Unfolded Protein Response (UPR). The UPR is the primary signaling pathway that monitors the state of the ER folding environment. When the unfolded protein burden overwhelms the capacity of the ER quality control machinery, a state termed ER stress, sensor proteins detect accumulation of misfolded peptides and trigger the UPR transcriptional response. The UPR, which is conserved from yeast to mammals, consists of an ensemble of complex signaling pathways that aims at adapting the ER to the new misfolded protein load. To determine how different factors impact the ER folding environment, various tools and assays have been developed. In this review, we discuss recent advances in live cell imaging reporters and model systems that enable researchers to monitor changes in the unfolded secretory protein burden and activation of the UPR and its associated signaling pathways.

show abstract

Multicolor and Electron Microscopic Imaging of Connexin Trafficking

Cited by 865 publications

References 26 publications

Connexins, cell motility, and the cytoskeleton

Connexins, cell motility, and the cytoskeleton

Gap junctions in olfactory neurons modulate olfactory sensitivity

Approaches to imaging unfolded secretory protein stress in living cells

Contact Info

Product

Resources

About