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Connexins

Connexins are a family of transmembrane proteins that form gap junctions, allowing for direct communication between cells.
These channels enable the exchange of small molecules, ions, and signaling molecules, playing crucial roles in tissue homeostasis, development, and disease processes.
PubCompare.ai's AI-powered platform can help researchers optimize their Connexins studies by easily identifying the best protocols from literature, preprints, and patents.
Leverage AI-insigths to streamline your workflow and take your Connexins research to the next level.

Most cited protocols related to «Connexins»

1
Dye Loading Protocols for Connexin-Expressing HeLa Cells
Cited 7 times
Connexin expressing HeLa cells were plated on cover slips. A coverslip was placed in a small flow chamber and the cells were exposed to either: control aCSF with 200 µM carboxyfluorescein for 10 min; isohydric hypercapnic aCSF with 200 µM carboxyfluorescein for 10 min; or zero Ca2+, 1 mM EGTA-containing aCSF plus 200 µM carboxyfluorescein for 10 min. This was followed by control aCSF plus 200 µM carboxyfluorescein for 5 min and then thorough washing for 30 min with control aCSF. These protocols are summarized in Figure 7.10.7554/eLife.01213.017Dye loading protocols.

The control background loading tests for any potential CO2-insensitive pathways of dye loading that are constitutively active in the HeLa cells. Hypercapnic dye loading uses the 50 mM HCO3− aCSF to test CO2-sensitive loading under conditions of isohydric hypercapnia (PCO2 55 mmHg). The zero Ca2+ positive control tests for the presence of functional hemichannels in those cases where the misexpressed hemichannels exhibit no sensitivity to CO2.

DOI:http://dx.doi.org/10.7554/eLife.01213.017

The cells were then imaged by epifluorescence (Scientifica Slice Scope (Scientifica Ltd, Uckfield, UK), Cairn Research OptoLED illumination (Cairn Research Limited, Faversham, UK), 60x water Olympus immersion objective, NA 1.0 (Scientifica), Hamamatsu ImageEM EMCCD camera (Hamamatsu Photonics K.K., Japan), Metafluor software (Cairn Research)). Using ImageJ, the extent of dye loading was measured by drawing a region of interest (ROI) around individual cells and calculating the mean pixel intensity for the ROI. The mean pixel intensity of the background fluorescence was also measured in a representative ROI, and this value was subtracted from the measures obtained from the cells. All of the images displayed in the figures reflect this procedure in that the mean intensity of the pixels in a representative background ROI has been subtracted from every pixel of the image. At least 40 cells were measured in each condition, and the mean pixel intensities plotted as cumulative probability distributions.
For the dye loading experiments, the median pixel intensities of the control and CO2 dye loading conditions (minimum of five independent repetitions) were compared by a Kruskal Wallace ANOVA and pairwise comparions by the Mann-Whitney test. The false discovery rate procedure (Curran-Everett, 2000 (link)) was used to determine whether the multiple pairwise comparisons remained significant.
Meigh L., Greenhalgh S.A., Rodgers T.L., Cann M.J., Roper D.I, & Dale N. (2013). CO2 directly modulates connexin 26 by formation of carbamate bridges between subunits. eLife, 2, e01213.
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Publication 2013
Bicarbonates carboxyfluorescein Cells Connexins Egtazic Acid Fluorescence HeLa Cells Hypersensitivity Light neuro-oncological ventral antigen 2, human Submersion
2
Hypercapnic stress response in Cx26A88V transfected HeLa cells
Cited 5 times
Coverslips plated with HeLa cells transiently transfected with Cx26A88V were exposed to Hypercapnic aCSF (55 mmHg or 70 mmHg) containing 200 µM CBF for 10 min. This was followed by control aCSF with 200 µM CBF for 5 min and a 30 min wash with control aCSF to ensure that all dye is removed from the outside of the cells.
A control comparison was used to establish any baseline loading occurring in the absence of a stimulus. HeLa cells expressing Cx26A88V were exposed to 200 µM CBF in control aCSF for 15 min, followed by 30 min of washing.
A zero Ca2+ positive control was also performed to ensure functional connexin hemichannels were being expressed. Cx26A88V expressing HeLa cells were exposed to 200 µM CBF in zero Ca2+ aCSF for 10 min. This was followed by control aCSF with 200 µM CBF for 5 min and 30 min of washing with aCSF.
Meigh L., Hussain N., Mulkey D.K, & Dale N. (2014). Connexin26 hemichannels with a mutation that causes KID syndrome in humans lack sensitivity to CO2. eLife, 3, e04249.
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Publication 2014
Cells Connexins HeLa Cells
3
Quantifying Connexin Protein Levels
Cited 5 times

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Publication 2009
anti-IgG Antibodies Cells Connexins GJA1 protein, human Goat Immobilon P Milk, Cow's Peroxidase polyacrylamide gels Proteins Rabbits Saline Solution SDS-PAGE Tail Technique, Dilution Tissue, Membrane Tissues
4
Quantifying Connexin Expression by qPCR
Cited 5 times

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Publication 2009
Cells Connexins DNA, Complementary Enzymes GAPDH protein, human Genetic Profile GJA1 protein, human Mus Oligonucleotide Primers Reverse Transcription SYBR Green I Transcription, Genetic trizol
5
Modeling Atrial Myocyte-Myofibroblast Interactions
Cited 4 times
Mathematical modeling was done using a combination of (i) the Koivumäki et al. model (2011 (link)) of the human atrial action potential and intracellular [Ca2+]i changes, and (ii) the “active” model of the human atrial myofibroblast described in detail in Maleckar et al. (2009a (link),b (link)). For our simulations, the mathematical expression for the inwardly rectifying background K+ current, IK1, in the fibroblast originally developed by MacCannell et al. (2007 (link)) was modified. This was done so that this important non-linear background current, and its contribution to the fibroblast resting potential could be simulated more accurately. Specifically, the equation for IK1 in Nygren et al. (Nygren and Giles, 2000 (link)) was used, and IK1 current density was scaled for the size (capacitance) of the fibroblast. This adjustment required a directly related small change in the size of the Na+/K+ pump current.
Three variants of this myofibroblast/atrial myocyte model were used in the simulations shown in Figures 6–8:
The illustrations in Figures 6–9 were generated by using these myocyte/myofibroblast models, assuming connexin-mediated electrotonic cell-to-cell communication with an assumed gap junctional conductance of 0.5 nS. This value corresponds to the lower end of those measured between fibroblast-myocyte pairs in vitro (Maleckar et al., 2009a (link),b (link)).
In addition to a 1:1 myocyte-myofibroblast coupling scheme, two slightly more complex sets of starting conditions were explored in these simulations. Either 3 or 9 myofibroblasts were connected in series to 1 myocyte. Based on our findings that only approximately 30–50% of the isolated myofibroblasts express a measurable Na+ current, only approximately one third of these myofibroblasts models were programmed to express a Na+ current with the properties in Figures 1–4 (model variant V1).
Koivumäki J.T., Clark R.B., Belke D., Kondo C., Fedak P.W., Maleckar M.M, & Giles W.R. (2014). Na+ current expression in human atrial myofibroblasts: identity and functional roles. Frontiers in Physiology, 5, 275.
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Publication 2014
Action Potentials Connexins Fibroblasts Heart Atrium Homo sapiens Intercellular Junctions Membrane Potentials Muscle Cells Myofibroblasts Protoplasm

Most recents protocols related to «Connexins»

1
Comprehensive Neural Interaction Database
Cited 1 time
NeuronChatDB is curated from existing databases (including KEGG53 (link) and IUPHAR/BPS Guide to PHARMACOLOGY54 (link)) and literature (e.g., neuropeptide interactions are from the reference16 (link)), and contains neural-specific intercellular molecular interactions for both mouse and human. There are 373 entries in total. Each entry of NeuronChatDB represents an interaction pair, including one ligand and a cognate target as well as genes related to them. The ligands include small-molecule neurotransmitters, neuropeptides, gap junction proteins, gasotransmitters and synaptic adhesion molecules: small-molecule neurotransmitters include glutamate (Glu), GABA, glycine (Gly), acetylcholine (ACh), serotonin (5-HT), dopamine (DA), epinephrine (Epi) and norepinephrine (NE); gasotransmitters include carbon monoxide (CO) and Nitric oxide (NO); synaptic adhesion molecules refer to neurexins (regarded as the ligand) and neuroligins (regarded as the target). The targets are typically but not limited to receptors. For example, the target proteins for neurotransmitters can also be uptake transporters or deactivating enzymes; the target proteins for gap junction proteins are other compatible gap junction proteins. For non-peptide neurotransmitters, corresponding synthesizing enzymes and/or vesicular transporters are included in the entry; for heteromeric receptors that contain multiple different subunits, corresponding subunits are split into different entries with the same ligand. To be compatible with the inference model of NeuronChat, for the non-peptide neurotransmitters, related genes including vesicular transporters and synthesizing enzymes responsible for different catalyzing steps are annotated into separate groups.
Zhao W., Johnston K.G., Ren H., Xu X, & Nie Q. (2023). Inferring neuron-neuron communications from single-cell transcriptomics through NeuronChat. Nature Communications, 14, 1128.
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Publication 2023
Acetylcholine Cell Adhesion Molecules Connexins Dopamine Enzymes Epinephrine gamma Aminobutyric Acid Gasotransmitters Genes Glutamate Glycine Homo sapiens Ligands Membrane Transport Proteins Monoxide, Carbon Nervous Mice Neuropeptides Neurotransmitters Norepinephrine Oxide, Nitric Peptides Proteins Protein Subunits Protein Targeting, Cellular
2
Rat Epidermal Keratinocytes for Tissue Engineering
Rat epidermal keratinocytes (REKs) were originally obtained from Dr. Vincent Hascall (Cleveland Clinic, Cleveland, Ohio, United States). REKs are spontaneously immortal but retain the capacity to form an organotypic rat epidermis when grown in an air-liquid interface (Maher et al., 2005 (link); Au et al., 2020 (link)). REKs express transcripts for eight connexins, but only express high levels of Cx43 protein with trace levels of Cx26 detectable in overgrown cells (Maher et al., 2005 (link); Au et al., 2020 (link)). Therefore, Cx43-ablated REKs (Cx43KO REKs) were generated using CRISPR/Cas9 technology as described and characterized earlier (Au et al., 2020 (link)). REKs, Cx43KO REKs, and normal rat kidney (NRK) cells were grown in sterile T25 (Fisher Brand, Burlington, ON, Canada, Cat# FB012935) or T75 flasks (Fisher Brand, Cat# FBO12937) containing Dulbecco’s modified Eagle’s medium (DMEM; ThermoFisher, Burlington, ON, Canada, Cat# 11960) fortified with 10% (v/v) fetal bovine serum (FBS; ThermoFisher, Cat# 12483-020), 2 mM L-glutamine (ThermoFisher, Cat# 25030-081), and 100 U/mL penicillin/streptomycin (ThermoFisher, Cat# 15140-122). Cells were cultured at 5% CO2 and 37°C (unless otherwise stated) and subcultured when 80%–90% confluent into 35 mm plastic culture dishes or 35 mm 6-well plates using 0.25% (w/v) trypsin-ethylene-diamine-tetraacetic acid (Trypsin-EDTA; ThermoFisher, Cat# 25200056). Connexin-deficient Neuro-2a (N2a) cells were used for electrophysiology experiments and cultured under the same conditions outlined above.
Lucaciu S.A., Figliuzzi R., Neumann R., Nazarali S., Del Sordo L., Leighton S.E., Hauser A., Shao Q., Johnston D., Bai D, & Laird D.W. (2023). GJB4 variants linked to skin disease exhibit a trafficking deficiency en route to gap junction formation that can be restored by co-expression of select connexins. Frontiers in Cell and Developmental Biology, 11, 1073805.
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Publication 2023
Cells Clustered Regularly Interspaced Short Palindromic Repeats Connexins Eagle Edetic Acid Epidermis GJA1 protein, human GJB2 protein, human Glutamine Hyperostosis, Diffuse Idiopathic Skeletal Keratinocyte Kidney Penicillins Proteins Sterility, Reproductive Streptomycin Trypsin
3
Improving Protein Folding and Trafficking
To assess the ability of sub-physiological temperature incubation to improve protein folding and trafficking, cells expressing Cx30.3 or EKVP-linked mutants were incubated at 26°C for 48 h, fixed and subsequently immunolabelled as described earlier. To assess the potential for rescue of EKVP-linked mutants with chemical chaperones, connexin or mutant expressing cells were treated with 5 mM 4-phenylbutyrate (4-PBA; MilliporeSigma, Cat# 1716-12-7), or 500 μM tauroursodeoxycholic acid (TUDCA; MilliporeSigma, Cat# 14605-22-2). Drug treated cells were fixed and immunolabelled as described earlier. Connexin or mutant expressing cells treated with 10% (v/v) glycerol Caledon Laboratories, Georgetown, ON, Canada Cat# 5350-1) for 24 and 48 h were grown in glass-bottom 35 mm dishes and live-imaged as described earlier.
Lucaciu S.A., Figliuzzi R., Neumann R., Nazarali S., Del Sordo L., Leighton S.E., Hauser A., Shao Q., Johnston D., Bai D, & Laird D.W. (2023). GJB4 variants linked to skin disease exhibit a trafficking deficiency en route to gap junction formation that can be restored by co-expression of select connexins. Frontiers in Cell and Developmental Biology, 11, 1073805.
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Publication 2023
4-PBA 4-phenylbutyrate Cells Connexins Erythrokeratodermia Variabilis GIT1 protein, human GIT2 protein, human Glycerin Hyperostosis, Diffuse Idiopathic Skeletal Molecular Chaperones Pharmaceutical Preparations physiology tauroursodeoxycholic acid
4
Comparative Sequence Analysis of Vertebrate Cx30.3 and β-Connexins
The OMA (Ortholog MAtrix) phylogenomic database (https://omabrowser.org/) was queried for the available primary sequences of Cx30.3 and all seven β-connexins from various vertebrate species. Multi sequence alignment was then performed using Clustal Omega in Jalview version 2.11.2.0 (https://jalview.org/). Sequence logos were then generated for the span of nine amino acids flanking the G12, T85, or F189 residues using the WebLogo sequence logo generator (https://weblogo.berkeley.edu/) (Schneider and Stephens, 1990 (link); Crooks et al., 2004 (link); Bai, 2016 (link)).
Lucaciu S.A., Figliuzzi R., Neumann R., Nazarali S., Del Sordo L., Leighton S.E., Hauser A., Shao Q., Johnston D., Bai D, & Laird D.W. (2023). GJB4 variants linked to skin disease exhibit a trafficking deficiency en route to gap junction formation that can be restored by co-expression of select connexins. Frontiers in Cell and Developmental Biology, 11, 1073805.
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Publication 2023
Amino Acids Connexins Sequence Alignment Vertebrates
5
Measuring Connexin Hemichannel Activity
Two days after injection of 20 ng connexin RNA, the whole cell membrane conductance of control and Cx26- or Cx30-expressing oocytes were measured by a two-electrode voltage clamp. As we had demonstrated that Cx26 and Cx30 hemichannel conductance could reach a steady state after 10 min equilibrium in Ca2+-free KCl solution (not shown), the conductances were measured 10 min after transferring from L15 medium to KCl solution. After conductance measurement, oocytes were injected with 32 nL of either 10 mM Alexa dyes or 1.25 mCi/mL 35S-labeled 10 μM ATP-γ-S (Perkin-Elmer, Austin, TX, USA) in L15 medium and then washed three times with L15 medium and Ca2+-free KCl solution once (85 mM KCl, 1 mM MgCl2 and 5 mM HEPES at pH 7.4). They were then transferred into Eppendorf tubes with 110 μL KCl, Ca2+-free solution. After 30 min, 100 μL medium was collected and mixed with 900 μL water. The remaining oocyte was rinsed once with 110 uL KCl and then lysed in 0.1% SDS lysis buffer. Alex dyes released to the media (M) were measured at the appropriate wavelength on a Fluromax-3 fluorimeter (Horiba, Japan), and ATP released to the media (M) was measured by scintillation counting on an LS6500 Beckman instrument (Beckman Coulter Life Sciences, Indianapolis, IN, USA) after mixing the samples with 10 mL UniverSol scintillation emulsifier (MP Biomedicals LLC, Irvine, CA, USA). The lysed oocyte (O) was diluted to the same volume and similarly measured in order to generate the M/O (media/oocyte) ratio of fluorescence or radiation.
Xu J, & Nicholson B.J. (2023). Divergence between Hemichannel and Gap Junction Permeabilities of Connexin 30 and 26. Life, 13(2), 390.
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Publication 2023
austin Buffers Connexins Dyes Fluorescence GJB2 protein, human HEPES Magnesium Chloride Oocytes Radiation

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More about "Connexins"

Connexins are a family of transmembrane proteins that form gap junctions, enabling direct communication between cells.
These channels facilitate the exchange of small molecules, ions, and signaling compounds, playing crucial roles in tissue homeostasis, development, and disease processes.
Researchers can optimize their Connexins studies by leveraging PubCompare.ai's AI-powered platform to easily identify the best protocols from literature, preprints, and patents.
Connexins, also known as gap junction proteins or GJPs, are a group of integral membrane proteins that assemble to form intercellular channels called gap junctions.
These channels allow for the direct exchange of ions, small molecules, and signaling messengers between neighboring cells, enabling coordinated cellular activities and tissue-level homeostasis.
Connexin research is crucial for understanding a wide range of biological processes, from embryonic development and tissue differentiation to wound healing and disease pathogenesis.
Studying Connexins can provide insights into the regulation of cellular communication, metabolic coupling, and signaling cascades that govern tissue function and homeostasis.
Researchers can leverage PubCompare.ai's AI-powered platform to streamline their Connexin studies.
The platform's intelligent comparison tools can help identify the most effective protocols, techniques, and products from the scientific literature, preprints, and patent databases.
By utilizing PubCompare.ai's AI-driven insights, researchers can optimize their experimental designs, choose the most appropriate reagents (e.g., SP6 mMessage mMachine, DMEM, FBS, Lipofectamine, Penicillin/streptomycin, Carbenoxolone, PBluescript II, AxioVision software 4.8, Accutase, X-tremeGENE HP), and enhance the efficiency of their Connexin research workflows.
Harness the power of PubCompare.ai to take your Connexin studies to the next level.
Discover the latest advancements, identify best practices, and streamline your research processes to accelerate your discoveries in this critical field of cell biology and biomedical research.