cx43  (Alomone Labs)


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    Name:
    Anti Connexin 43 Antibody
    Description:
    Anti Connexin 43 Antibody is directed against an epitope of human Connexin 43 Anti Connexin 43 Antibody ACC 201 can be used in western blot immunohistochemistry and immunocytochemistry applications It has been designed to recognize Cx43 from human rat and mouse samples
    Catalog Number:
    ACC-201
    Price:
    397.0
    Category:
    Primary Antibody
    Applications:
    Immunocytochemistry, Immunofluorescence, Immunohistochemistry, Western Blot
    Purity:
    Affinity purified on immobilized antigen.
    Immunogen:
    Synthetic peptide
    Size:
    25 mcl
    Antibody Type:
    Polyclonal Primary Antibodies
    Format:
    Lyophilized Powder
    Host:
    Rabbit
    Isotype:
    Rabbit IgG
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    Structured Review

    Alomone Labs cx43
    Anti Connexin 43 Antibody
    Anti Connexin 43 Antibody is directed against an epitope of human Connexin 43 Anti Connexin 43 Antibody ACC 201 can be used in western blot immunohistochemistry and immunocytochemistry applications It has been designed to recognize Cx43 from human rat and mouse samples
    https://www.bioz.com/result/cx43/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cx43 - by Bioz Stars, 2021-09
    94/100 stars

    Images

    1) Product Images from "Blockade of Glial Connexin 43 Hemichannels Reduces Food Intake"

    Article Title: Blockade of Glial Connexin 43 Hemichannels Reduces Food Intake

    Journal: Cells

    doi: 10.3390/cells9112387

    ( A ): Representative images depicting ethidium bromide (EtBr) uptake via hemichannels in hypothalamic (ARC) slices treated with either artificial cerebrospinal fluid (aCSF), 200 μM carbenoxolone (Cbx) or 100 µM the connexin-43 (Cx43) hemichannel blocker TAT-GAP19 for 15 min, then exposed to EtBr for 10 min. After fixation, slices were submitted to Glial Fibrillary Acidic Protein (GFAP) immunostaining and Hoechst labelling. Scale bar: 15 µm. ( B ): Quantification of EtBr fluorescence measured in GFAP+ astrocytes. n = 5, with each n representing the average of three replicates. ** p
    Figure Legend Snippet: ( A ): Representative images depicting ethidium bromide (EtBr) uptake via hemichannels in hypothalamic (ARC) slices treated with either artificial cerebrospinal fluid (aCSF), 200 μM carbenoxolone (Cbx) or 100 µM the connexin-43 (Cx43) hemichannel blocker TAT-GAP19 for 15 min, then exposed to EtBr for 10 min. After fixation, slices were submitted to Glial Fibrillary Acidic Protein (GFAP) immunostaining and Hoechst labelling. Scale bar: 15 µm. ( B ): Quantification of EtBr fluorescence measured in GFAP+ astrocytes. n = 5, with each n representing the average of three replicates. ** p

    Techniques Used: Immunostaining, Fluorescence

    ( A , B ): Connexin-43 (Cx43) and Glial Fibrillary Acidic Protein (GFAP) mRNA expression (normalized with Glyceraldehyde-3-phosphate dehydrogenase gene, GAPDH) was assessed by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) from the hypothalamus ( A ) and dorsal vagal complex (DVC) ( B ) of control ad libitum ( n = 14), 24 h fasted ( n = 13), 24 h fasted plus 2 h refeed ( n = 8) or plus 4 h refeed ( n = 8) mice. ( C ): Representative Western blot analysis of phosphorylated Cx43 (pS368) and Cx43 expression in the hypothalamus of control ad libitum ( n = 5) and 24 h fasted ( n = 5) mice. ( D , E ): Cx43 and GFAP mRNA expressions (normalized with GAPDH) were assessed by Reverse Transcription quantitative Polymerase Chain Reaction (RT-qPCR) from the hypothalamus ( D ) and dorsal vagal complex (DVC) ( D ) of Normal Chow (NC) ( n = 9) and High Fat Diet (HFD) fed ( n = 9) mice. ( F ): Representative Western blot analysis and quantification of phosphorylated Cx43 (pS368) and Cx43 expression in the hypothalamus of NC ( n = 9) and HFD-fed ( n = 9) mice. A two-way analysis of variance test was performed between different experimental groups * p
    Figure Legend Snippet: ( A , B ): Connexin-43 (Cx43) and Glial Fibrillary Acidic Protein (GFAP) mRNA expression (normalized with Glyceraldehyde-3-phosphate dehydrogenase gene, GAPDH) was assessed by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) from the hypothalamus ( A ) and dorsal vagal complex (DVC) ( B ) of control ad libitum ( n = 14), 24 h fasted ( n = 13), 24 h fasted plus 2 h refeed ( n = 8) or plus 4 h refeed ( n = 8) mice. ( C ): Representative Western blot analysis of phosphorylated Cx43 (pS368) and Cx43 expression in the hypothalamus of control ad libitum ( n = 5) and 24 h fasted ( n = 5) mice. ( D , E ): Cx43 and GFAP mRNA expressions (normalized with GAPDH) were assessed by Reverse Transcription quantitative Polymerase Chain Reaction (RT-qPCR) from the hypothalamus ( D ) and dorsal vagal complex (DVC) ( D ) of Normal Chow (NC) ( n = 9) and High Fat Diet (HFD) fed ( n = 9) mice. ( F ): Representative Western blot analysis and quantification of phosphorylated Cx43 (pS368) and Cx43 expression in the hypothalamus of NC ( n = 9) and HFD-fed ( n = 9) mice. A two-way analysis of variance test was performed between different experimental groups * p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Mouse Assay, Western Blot

    ( A ): Confocal images of connexin-43 (Cx43) (red) and bassoon (green) double labeling immunohistochemistry performed within the nucleus tractus solitarius (NTS). Scale bar: 5 μm. Inset: Arrows show Cx43 labelling in close vicinity of synapses labelled by presynaptic bassoon protein. Scale bar 1 µm. ( B ): An example of a voxel (4 × 4 µm) used for the quantification of Cx43/bassoon appositions. Scale bar: 0.5 µm. ( C ): Twelve z-stack images defining the voxel illustrated in B and allowing the identification of Cx43/bassoon appositions. ( D ): Stereological estimation of Cx43/bassoon appositions in the NTS (85.13%) and arcuate nucleus of the hypothalamus (ARC) (80.46%) nuclei. ( E ): Table gathering the different stereological values acquired or estimated for each studied animal.
    Figure Legend Snippet: ( A ): Confocal images of connexin-43 (Cx43) (red) and bassoon (green) double labeling immunohistochemistry performed within the nucleus tractus solitarius (NTS). Scale bar: 5 μm. Inset: Arrows show Cx43 labelling in close vicinity of synapses labelled by presynaptic bassoon protein. Scale bar 1 µm. ( B ): An example of a voxel (4 × 4 µm) used for the quantification of Cx43/bassoon appositions. Scale bar: 0.5 µm. ( C ): Twelve z-stack images defining the voxel illustrated in B and allowing the identification of Cx43/bassoon appositions. ( D ): Stereological estimation of Cx43/bassoon appositions in the NTS (85.13%) and arcuate nucleus of the hypothalamus (ARC) (80.46%) nuclei. ( E ): Table gathering the different stereological values acquired or estimated for each studied animal.

    Techniques Used: Labeling, Immunohistochemistry

    ( A ): Cumulative food intake (g), measured over a 24 h period, of mice having received an i.c.v. injection of either vehicle (Sodium Chloride (NaCl) 0.9%) or TAT-GAP19 (12.5 or 25 µg). The TAT-GAP19-I14A, which includes a point mutation making it ineffective on connexin 43 hemichannels (Cx43 HCs) activity was tested on food intake at the dose of 25 µg. ( B ): Graph showing meal size over 24 h after i.c.v. injection of vehicle or TAT-GAP19 25 µg. Note that each bar represents a meal and its width the meal duration. The dark period is represented by the shaded box. ( C – F ): Meal microstructure analysis performed during the dark period for mice that received either vehicle or TAT-GAP19 25 µg allowed quantification of meals number ( C ), meals size (in mg of food, ( D )), meal duration (in min, ( E )) and interprandial period (in min, average intervals between meals, ( F – G )): Measure of energy expenditure (in Kcal/day-1/kg 0.75 ) within 12 h after injection of 0.9% NaCl vehicle ( n = 7) or 50 µg TAT-GAP19 ( n = 7). ( H , I ): Measure of total spontaneous activity (in Arbitrary Unit, UA; ( H ) and cumulative spontaneous activity ( I ) within 12 h for the 2 groups. ns: no significant difference; * p
    Figure Legend Snippet: ( A ): Cumulative food intake (g), measured over a 24 h period, of mice having received an i.c.v. injection of either vehicle (Sodium Chloride (NaCl) 0.9%) or TAT-GAP19 (12.5 or 25 µg). The TAT-GAP19-I14A, which includes a point mutation making it ineffective on connexin 43 hemichannels (Cx43 HCs) activity was tested on food intake at the dose of 25 µg. ( B ): Graph showing meal size over 24 h after i.c.v. injection of vehicle or TAT-GAP19 25 µg. Note that each bar represents a meal and its width the meal duration. The dark period is represented by the shaded box. ( C – F ): Meal microstructure analysis performed during the dark period for mice that received either vehicle or TAT-GAP19 25 µg allowed quantification of meals number ( C ), meals size (in mg of food, ( D )), meal duration (in min, ( E )) and interprandial period (in min, average intervals between meals, ( F – G )): Measure of energy expenditure (in Kcal/day-1/kg 0.75 ) within 12 h after injection of 0.9% NaCl vehicle ( n = 7) or 50 µg TAT-GAP19 ( n = 7). ( H , I ): Measure of total spontaneous activity (in Arbitrary Unit, UA; ( H ) and cumulative spontaneous activity ( I ) within 12 h for the 2 groups. ns: no significant difference; * p

    Techniques Used: Mouse Assay, Injection, Mutagenesis, Activity Assay

    Related Articles

    Staining:

    Article Title: Domain-specific distribution of gap junctions defines cellular coupling to establish a vascular relay in the retina
    Article Snippet: .. In addition, when the Alomone’s Cx43 antibody was preincubated with the control peptide antigen, the staining in the mouse retina was eliminated ( ). ..

    Microscopy:

    Article Title: Beneficial effect of voluntary physical exercise in Plakophilin2 transgenic mice
    Article Snippet: .. The sections were labelled with anti-Cx43 (1:400, Alomone Labs, Jerusalem, Israel) and Alexa488 (1:500, Invitrogen) and subsequently analyzed with a confocal microscope. ..

    Laser Capture Microdissection:

    Article Title: Development of the stria vascularis and potassium regulation in the human fetal cochlea: Insights into hereditary sensorineural hearing loss
    Article Snippet: .. The primary antibodies used in this study were mouse anti‐acetylated tubulin (aceTUBA, 1:500, T6793, Sigma), rabbit anti‐collagen type IV (COL4, 1:200, AB748, Chemicon), rabbit anti‐fibronectin (FN, 1:400, F3648, Sigma‐Aldrich), rabbit anti‐GJA1 (1:1000, C6219, Sigma), rabbit anti‐GJA1 (1:2000, ACC‐201, Alomone labs), rabbit anti‐GJB2 (1:100, ACC‐212, Alomone labs), rabbit anti‐GJB6 (1:100, PA511640, Thermo Scientific), rabbit anti‐GJE1 (1:1000, NBP2–14051, Novus biologicals), goat anti‐KCNJ10 (1:100, NBP1–70371), rabbit anti‐KCNQ1 (1:200, ab65092, Abcam), rabbit anti‐KCNQ1 (1:100, APC‐022, Alomone labs), rabbit anti‐KIT (1:100, A4502, Dako), rabbit anti‐laminin (LAM, 1:200, Z009701, Dako), rabbit anti‐melan‐A (1:500, NBP1–30151, Novus), mouse anti‐microphthalmia‐associated transcription factor (MITF, 1:100, M3621, Dako), mouse anti‐Na+ /K+ ‐ATPase α1 (ATP1A1, 1:200, α6F, Developmental Studies Hybridoma Bank), rabbit anti‐solute family carrier 2, member 1 (SLC2A1, 1:500, ab15309, Abcam), and goat anti‐SOX10 (1:50, sc‐17342, Santa Cruz Biotechnology). ..

    Labeling:

    Article Title: Astrocyte remodeling without gliosis precedes optic nerve Axonopathy
    Article Snippet: .. We labeled 10 μm cryosections with the following antibodies: anti-glial fibrillary acidic protein (GFAP; EMD Millipore, Billerica, MA, 1:500), and anti-Connexin-43 (Cx43; Alomone Labs, Jerusalem, Israel, 1:250). ..

    other:

    Article Title: Directed and systematic differentiation of cardiovascular cells from mouse induced pluripotent stem cells.
    Article Snippet: Anti-Kir2.1 (1:200) and anticonnexin 43 (1:200) antibodies were from Alomone (Israel) and Invitrogen (Carlsbad, Calif), respectively.

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  • 94
    Alomone Labs cx43 rabbit primary antibody
    ( A ): Representative images depicting ethidium bromide (EtBr) uptake via hemichannels in hypothalamic (ARC) slices treated with either artificial cerebrospinal fluid (aCSF), 200 μM carbenoxolone (Cbx) or 100 µM the connexin-43 <t>(Cx43)</t> hemichannel blocker TAT-GAP19 for 15 min, then exposed to EtBr for 10 min. After fixation, slices were submitted to Glial Fibrillary Acidic Protein (GFAP) immunostaining and Hoechst labelling. Scale bar: 15 µm. ( B ): Quantification of EtBr fluorescence measured in GFAP+ astrocytes. n = 5, with each n representing the average of three replicates. ** p
    Cx43 Rabbit Primary Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cx43 rabbit primary antibody/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cx43 rabbit primary antibody - by Bioz Stars, 2021-09
    94/100 stars
      Buy from Supplier

    94
    Alomone Labs connexin 43 cx43
    Histological analysis of cardiac tissue after long-term training. ( A ) Representative images for sedentary, 8 weeks of running and 16 weeks of running tissue from wt (upper row) and PKP2 +/- mice (lower row) are shown for Trichrome-stained ventricular slices. No change of fibrous tissue could be detected in either genotype or in response to long-term training. ( B ) The oil stain in frozen sections from cardiac tissue with oil red did not reveal an increase in fatty replacements. ( C ) We labeled the gap junctions with a <t>Cx43</t> specific antibody and analyzed the images by confocal microscopy. The connexins were predominantly located at the intercalated discs as seen in the representative images shown. There was no shift in subcellular distribution due to genotype or long-term training observed. The single data points indicate the number of animals analyzed.
    Connexin 43 Cx43, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/connexin 43 cx43/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    connexin 43 cx43 - by Bioz Stars, 2021-09
    94/100 stars
      Buy from Supplier

    Image Search Results


    ( A ): Representative images depicting ethidium bromide (EtBr) uptake via hemichannels in hypothalamic (ARC) slices treated with either artificial cerebrospinal fluid (aCSF), 200 μM carbenoxolone (Cbx) or 100 µM the connexin-43 (Cx43) hemichannel blocker TAT-GAP19 for 15 min, then exposed to EtBr for 10 min. After fixation, slices were submitted to Glial Fibrillary Acidic Protein (GFAP) immunostaining and Hoechst labelling. Scale bar: 15 µm. ( B ): Quantification of EtBr fluorescence measured in GFAP+ astrocytes. n = 5, with each n representing the average of three replicates. ** p

    Journal: Cells

    Article Title: Blockade of Glial Connexin 43 Hemichannels Reduces Food Intake

    doi: 10.3390/cells9112387

    Figure Lengend Snippet: ( A ): Representative images depicting ethidium bromide (EtBr) uptake via hemichannels in hypothalamic (ARC) slices treated with either artificial cerebrospinal fluid (aCSF), 200 μM carbenoxolone (Cbx) or 100 µM the connexin-43 (Cx43) hemichannel blocker TAT-GAP19 for 15 min, then exposed to EtBr for 10 min. After fixation, slices were submitted to Glial Fibrillary Acidic Protein (GFAP) immunostaining and Hoechst labelling. Scale bar: 15 µm. ( B ): Quantification of EtBr fluorescence measured in GFAP+ astrocytes. n = 5, with each n representing the average of three replicates. ** p

    Article Snippet: Once again, the same saturation solution was used before the Cx43 rabbit primary antibody (#ACC-201 Alomone Labs 1/3000, Jerusalem, Israel) and goat anti-rabbit-IgG secondary antibody (1/400) on the same conditions.

    Techniques: Immunostaining, Fluorescence

    ( A , B ): Connexin-43 (Cx43) and Glial Fibrillary Acidic Protein (GFAP) mRNA expression (normalized with Glyceraldehyde-3-phosphate dehydrogenase gene, GAPDH) was assessed by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) from the hypothalamus ( A ) and dorsal vagal complex (DVC) ( B ) of control ad libitum ( n = 14), 24 h fasted ( n = 13), 24 h fasted plus 2 h refeed ( n = 8) or plus 4 h refeed ( n = 8) mice. ( C ): Representative Western blot analysis of phosphorylated Cx43 (pS368) and Cx43 expression in the hypothalamus of control ad libitum ( n = 5) and 24 h fasted ( n = 5) mice. ( D , E ): Cx43 and GFAP mRNA expressions (normalized with GAPDH) were assessed by Reverse Transcription quantitative Polymerase Chain Reaction (RT-qPCR) from the hypothalamus ( D ) and dorsal vagal complex (DVC) ( D ) of Normal Chow (NC) ( n = 9) and High Fat Diet (HFD) fed ( n = 9) mice. ( F ): Representative Western blot analysis and quantification of phosphorylated Cx43 (pS368) and Cx43 expression in the hypothalamus of NC ( n = 9) and HFD-fed ( n = 9) mice. A two-way analysis of variance test was performed between different experimental groups * p

    Journal: Cells

    Article Title: Blockade of Glial Connexin 43 Hemichannels Reduces Food Intake

    doi: 10.3390/cells9112387

    Figure Lengend Snippet: ( A , B ): Connexin-43 (Cx43) and Glial Fibrillary Acidic Protein (GFAP) mRNA expression (normalized with Glyceraldehyde-3-phosphate dehydrogenase gene, GAPDH) was assessed by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) from the hypothalamus ( A ) and dorsal vagal complex (DVC) ( B ) of control ad libitum ( n = 14), 24 h fasted ( n = 13), 24 h fasted plus 2 h refeed ( n = 8) or plus 4 h refeed ( n = 8) mice. ( C ): Representative Western blot analysis of phosphorylated Cx43 (pS368) and Cx43 expression in the hypothalamus of control ad libitum ( n = 5) and 24 h fasted ( n = 5) mice. ( D , E ): Cx43 and GFAP mRNA expressions (normalized with GAPDH) were assessed by Reverse Transcription quantitative Polymerase Chain Reaction (RT-qPCR) from the hypothalamus ( D ) and dorsal vagal complex (DVC) ( D ) of Normal Chow (NC) ( n = 9) and High Fat Diet (HFD) fed ( n = 9) mice. ( F ): Representative Western blot analysis and quantification of phosphorylated Cx43 (pS368) and Cx43 expression in the hypothalamus of NC ( n = 9) and HFD-fed ( n = 9) mice. A two-way analysis of variance test was performed between different experimental groups * p

    Article Snippet: Once again, the same saturation solution was used before the Cx43 rabbit primary antibody (#ACC-201 Alomone Labs 1/3000, Jerusalem, Israel) and goat anti-rabbit-IgG secondary antibody (1/400) on the same conditions.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Mouse Assay, Western Blot

    ( A ): Confocal images of connexin-43 (Cx43) (red) and bassoon (green) double labeling immunohistochemistry performed within the nucleus tractus solitarius (NTS). Scale bar: 5 μm. Inset: Arrows show Cx43 labelling in close vicinity of synapses labelled by presynaptic bassoon protein. Scale bar 1 µm. ( B ): An example of a voxel (4 × 4 µm) used for the quantification of Cx43/bassoon appositions. Scale bar: 0.5 µm. ( C ): Twelve z-stack images defining the voxel illustrated in B and allowing the identification of Cx43/bassoon appositions. ( D ): Stereological estimation of Cx43/bassoon appositions in the NTS (85.13%) and arcuate nucleus of the hypothalamus (ARC) (80.46%) nuclei. ( E ): Table gathering the different stereological values acquired or estimated for each studied animal.

    Journal: Cells

    Article Title: Blockade of Glial Connexin 43 Hemichannels Reduces Food Intake

    doi: 10.3390/cells9112387

    Figure Lengend Snippet: ( A ): Confocal images of connexin-43 (Cx43) (red) and bassoon (green) double labeling immunohistochemistry performed within the nucleus tractus solitarius (NTS). Scale bar: 5 μm. Inset: Arrows show Cx43 labelling in close vicinity of synapses labelled by presynaptic bassoon protein. Scale bar 1 µm. ( B ): An example of a voxel (4 × 4 µm) used for the quantification of Cx43/bassoon appositions. Scale bar: 0.5 µm. ( C ): Twelve z-stack images defining the voxel illustrated in B and allowing the identification of Cx43/bassoon appositions. ( D ): Stereological estimation of Cx43/bassoon appositions in the NTS (85.13%) and arcuate nucleus of the hypothalamus (ARC) (80.46%) nuclei. ( E ): Table gathering the different stereological values acquired or estimated for each studied animal.

    Article Snippet: Once again, the same saturation solution was used before the Cx43 rabbit primary antibody (#ACC-201 Alomone Labs 1/3000, Jerusalem, Israel) and goat anti-rabbit-IgG secondary antibody (1/400) on the same conditions.

    Techniques: Labeling, Immunohistochemistry

    ( A ): Cumulative food intake (g), measured over a 24 h period, of mice having received an i.c.v. injection of either vehicle (Sodium Chloride (NaCl) 0.9%) or TAT-GAP19 (12.5 or 25 µg). The TAT-GAP19-I14A, which includes a point mutation making it ineffective on connexin 43 hemichannels (Cx43 HCs) activity was tested on food intake at the dose of 25 µg. ( B ): Graph showing meal size over 24 h after i.c.v. injection of vehicle or TAT-GAP19 25 µg. Note that each bar represents a meal and its width the meal duration. The dark period is represented by the shaded box. ( C – F ): Meal microstructure analysis performed during the dark period for mice that received either vehicle or TAT-GAP19 25 µg allowed quantification of meals number ( C ), meals size (in mg of food, ( D )), meal duration (in min, ( E )) and interprandial period (in min, average intervals between meals, ( F – G )): Measure of energy expenditure (in Kcal/day-1/kg 0.75 ) within 12 h after injection of 0.9% NaCl vehicle ( n = 7) or 50 µg TAT-GAP19 ( n = 7). ( H , I ): Measure of total spontaneous activity (in Arbitrary Unit, UA; ( H ) and cumulative spontaneous activity ( I ) within 12 h for the 2 groups. ns: no significant difference; * p

    Journal: Cells

    Article Title: Blockade of Glial Connexin 43 Hemichannels Reduces Food Intake

    doi: 10.3390/cells9112387

    Figure Lengend Snippet: ( A ): Cumulative food intake (g), measured over a 24 h period, of mice having received an i.c.v. injection of either vehicle (Sodium Chloride (NaCl) 0.9%) or TAT-GAP19 (12.5 or 25 µg). The TAT-GAP19-I14A, which includes a point mutation making it ineffective on connexin 43 hemichannels (Cx43 HCs) activity was tested on food intake at the dose of 25 µg. ( B ): Graph showing meal size over 24 h after i.c.v. injection of vehicle or TAT-GAP19 25 µg. Note that each bar represents a meal and its width the meal duration. The dark period is represented by the shaded box. ( C – F ): Meal microstructure analysis performed during the dark period for mice that received either vehicle or TAT-GAP19 25 µg allowed quantification of meals number ( C ), meals size (in mg of food, ( D )), meal duration (in min, ( E )) and interprandial period (in min, average intervals between meals, ( F – G )): Measure of energy expenditure (in Kcal/day-1/kg 0.75 ) within 12 h after injection of 0.9% NaCl vehicle ( n = 7) or 50 µg TAT-GAP19 ( n = 7). ( H , I ): Measure of total spontaneous activity (in Arbitrary Unit, UA; ( H ) and cumulative spontaneous activity ( I ) within 12 h for the 2 groups. ns: no significant difference; * p

    Article Snippet: Once again, the same saturation solution was used before the Cx43 rabbit primary antibody (#ACC-201 Alomone Labs 1/3000, Jerusalem, Israel) and goat anti-rabbit-IgG secondary antibody (1/400) on the same conditions.

    Techniques: Mouse Assay, Injection, Mutagenesis, Activity Assay

    Diminished anterograde transport in a sample of DBA/2 J nerves.  a  Top Left: Coronal section through superior colliculus of DBA/2 J mouse (between dashed white lines) following intravitreal injection of CTB (green). Bottom Left: Corresponding retinotopic map shows nearly depleted anterograde transport of CTB. Top Right: Coronal section through superior colliculus of D2 control mouse prepared as on left. Bottom Right: Corresponding retinotopic map shows a full complement of anterogradely transported CTB.  b  Transport of CTB from DBA/2 J eyes was near 20% (22.8 ± 5.5), significantly reduced from D2 control which is near 75% (74.896 ± 4.328) ( p  = 0.0015).  c  Confocal micrographs of proximal (left) and distal (right) DBA/2 J (top) and D2 control (bottom) optic nerves. Connexin 43 (Cx43, blue) and GFAP (red) colocalize, and both are elevated in proximal optic nerve.  d . Density of Cx43 (puncta/μm 2 ) in proximal segment of DBA/2 J nerves is significantly elevated compared to both distal DBA/2 J (*;  p  = 0.043) and proximal D2 nerves (#;  p  = 0.032)

    Journal: Acta Neuropathologica Communications

    Article Title: Astrocyte remodeling without gliosis precedes optic nerve Axonopathy

    doi: 10.1186/s40478-018-0542-0

    Figure Lengend Snippet: Diminished anterograde transport in a sample of DBA/2 J nerves. a Top Left: Coronal section through superior colliculus of DBA/2 J mouse (between dashed white lines) following intravitreal injection of CTB (green). Bottom Left: Corresponding retinotopic map shows nearly depleted anterograde transport of CTB. Top Right: Coronal section through superior colliculus of D2 control mouse prepared as on left. Bottom Right: Corresponding retinotopic map shows a full complement of anterogradely transported CTB. b Transport of CTB from DBA/2 J eyes was near 20% (22.8 ± 5.5), significantly reduced from D2 control which is near 75% (74.896 ± 4.328) ( p  = 0.0015). c Confocal micrographs of proximal (left) and distal (right) DBA/2 J (top) and D2 control (bottom) optic nerves. Connexin 43 (Cx43, blue) and GFAP (red) colocalize, and both are elevated in proximal optic nerve. d . Density of Cx43 (puncta/μm 2 ) in proximal segment of DBA/2 J nerves is significantly elevated compared to both distal DBA/2 J (*; p  = 0.043) and proximal D2 nerves (#; p  = 0.032)

    Article Snippet: We labeled 10 μm cryosections with the following antibodies: anti-glial fibrillary acidic protein (GFAP; EMD Millipore, Billerica, MA, 1:500), and anti-Connexin-43 (Cx43; Alomone Labs, Jerusalem, Israel, 1:250).

    Techniques: Injection, CtB Assay

    Cx43 is primarily localized to astrocytes and vascular cells. (a) Around an artery, majority of the large puncta was not directly associated with vascular cells. (b) A vertical view from the highlighted area in (a) was obtained by 90 0 rotation of the z-stack. On the right, fluorescence intensity profile for the three labeling from (b). (c–d) In the relay, large puncta were also not directly located on the blood vessels contrarily to the string-like structures which were enclosed by the mural cells (d panel and the intensity profile). (e–f) Around the finest capillaries, Cx43-puncta were mostly outside of the blood vessels. (h) In veins, putative labeling was not directly associated with mural cells. (i–j) Bright Cx43 puncta around blood vessels were localized to astroglia labeled for glial fibrillary acidic protein (GFAP). (k–m) Cx43-positive strings were associated with endothelial cells labeled for platelet endothelial cell adhesion molecule-1 (PECAM-1). Scale bar 5 μm.

    Journal: The Journal of comparative neurology

    Article Title: Domain-specific distribution of gap junctions defines cellular coupling to establish a vascular relay in the retina

    doi: 10.1002/cne.24699

    Figure Lengend Snippet: Cx43 is primarily localized to astrocytes and vascular cells. (a) Around an artery, majority of the large puncta was not directly associated with vascular cells. (b) A vertical view from the highlighted area in (a) was obtained by 90 0 rotation of the z-stack. On the right, fluorescence intensity profile for the three labeling from (b). (c–d) In the relay, large puncta were also not directly located on the blood vessels contrarily to the string-like structures which were enclosed by the mural cells (d panel and the intensity profile). (e–f) Around the finest capillaries, Cx43-puncta were mostly outside of the blood vessels. (h) In veins, putative labeling was not directly associated with mural cells. (i–j) Bright Cx43 puncta around blood vessels were localized to astroglia labeled for glial fibrillary acidic protein (GFAP). (k–m) Cx43-positive strings were associated with endothelial cells labeled for platelet endothelial cell adhesion molecule-1 (PECAM-1). Scale bar 5 μm.

    Article Snippet: In addition, when the Alomone’s Cx43 antibody was preincubated with the control peptide antigen, the staining in the mouse retina was eliminated ( ).

    Techniques: Fluorescence, Labeling

    Hierarchy within the vascular tree dictates distinct patterns of Cx43 clustering. (a) Cx43 is highly concentrated along retinal vasculature. Inset shows the arteries from the same area labeled for smooth muscle actin (SMA). Scale bar 200 μm. (b) In the superficial layer from area b in (a), majority of puncta were associated with vasculature: arteries, veins and capillaries and lower number of similar size puncta were present outside of the blood vessels. Scale bar 50 μm. (c) In the branches of major arteries from an area similar to the area c in (a), string-like structures running along the blood vessels were found in addition to the puncta. Scale bar 50 μm. (d) Density of the Cx43 puncta was the highest on veins and capillaries. The strings were found predominantly in relay regions connecting artery with capillaries. Density of Cx43 puncta outside of the blood vessels was significantly lower and no strings were detected. (e) Dendrogram created by cluster analysis of data from (d). (f) Schematic presentation of Cx43 expression pattern in retinal vasculature. (g) In a vertical section through the retina (created by rotation of a thin slit from the z-stack), the string-like structures from the superficial layer were extending along some capillaries into the intermediate and deep vascular layers. Scale bar 50 μm. (h) Distribution of Cx43 shows that Cx43 (puncta and strings combined) is predominantly expressed on the vasculature throughout the retina. The relays have the highest densities of Cx43-positive structures. The intermediate and deep layer capillaries above these relay zones also have the highest expression of Cx43 in string-like structures. The non-vascular expression in the superficial layer reflects the puncta outside of the blood vessels; no puncta of the same size and labeling intensity was detected in the intermediate and deep layers. Data are shown as average ± SD; arteries, vein and transitions were quantified in 6 samples from 6 mice; capillaries and non-vascular areas in n = 12 samples from n = 6 mice. ANOVA on ranks, * p = 0.0001.

    Journal: The Journal of comparative neurology

    Article Title: Domain-specific distribution of gap junctions defines cellular coupling to establish a vascular relay in the retina

    doi: 10.1002/cne.24699

    Figure Lengend Snippet: Hierarchy within the vascular tree dictates distinct patterns of Cx43 clustering. (a) Cx43 is highly concentrated along retinal vasculature. Inset shows the arteries from the same area labeled for smooth muscle actin (SMA). Scale bar 200 μm. (b) In the superficial layer from area b in (a), majority of puncta were associated with vasculature: arteries, veins and capillaries and lower number of similar size puncta were present outside of the blood vessels. Scale bar 50 μm. (c) In the branches of major arteries from an area similar to the area c in (a), string-like structures running along the blood vessels were found in addition to the puncta. Scale bar 50 μm. (d) Density of the Cx43 puncta was the highest on veins and capillaries. The strings were found predominantly in relay regions connecting artery with capillaries. Density of Cx43 puncta outside of the blood vessels was significantly lower and no strings were detected. (e) Dendrogram created by cluster analysis of data from (d). (f) Schematic presentation of Cx43 expression pattern in retinal vasculature. (g) In a vertical section through the retina (created by rotation of a thin slit from the z-stack), the string-like structures from the superficial layer were extending along some capillaries into the intermediate and deep vascular layers. Scale bar 50 μm. (h) Distribution of Cx43 shows that Cx43 (puncta and strings combined) is predominantly expressed on the vasculature throughout the retina. The relays have the highest densities of Cx43-positive structures. The intermediate and deep layer capillaries above these relay zones also have the highest expression of Cx43 in string-like structures. The non-vascular expression in the superficial layer reflects the puncta outside of the blood vessels; no puncta of the same size and labeling intensity was detected in the intermediate and deep layers. Data are shown as average ± SD; arteries, vein and transitions were quantified in 6 samples from 6 mice; capillaries and non-vascular areas in n = 12 samples from n = 6 mice. ANOVA on ranks, * p = 0.0001.

    Article Snippet: In addition, when the Alomone’s Cx43 antibody was preincubated with the control peptide antigen, the staining in the mouse retina was eliminated ( ).

    Techniques: Labeling, Expressing, Mouse Assay

    Validation and optimization of Connexin antibodies used in this study. (a–d) Cx37 labeling revealed just a few puncta (green) on blood vessels (blue) in pure PFA fixative (a) and entire outlines of contractile cells in carbodiimide-based fixatives (b and c). Ganglion cells were more visible in PFA fixative (asterisks); the lowest background was in carbodiimide. Pre-absorption of the antibody with a control peptide eliminated the labeling (d). (e–h) Cx40 (green) was associated with outlines of endothelial cells (blue) in all fixative with the less background in carbodiimide (f). Incubation of primary antibody eliminated the labeling (h) with the exception of labeled microglia (arrow). (i–k) Cx43s-positive puncta (green, Sigma antibody) were associated with vasculature in PFA fixative (i); in addition string-like structures along endothelial cell contacts (blue) became visible in carbodiimide fixative (j–k). (l) Omission of the primary antibody eliminated punctate and string-like structures; the microglia was labeled by the secondary antibody (arrow). (m–p) Cx43a produced identical pattern as Cx43s in carbodiimide containing fixatives (n, o). In pure PFA fixative, this antibody labeled well punctate and string-like structures; however the background was significantly higher (m). Incubation of the primary antibodies with control peptide eliminated the specific labeling (p). (q–s) Morphology of DsRed expressing pericytes was best preserved in PFA-containing fixatives (q and s); the DsRed labeling in pericyte processes was lost in pure carbodiimide fixative (q). (t) Summary of fixation procedure. Red crosses indicate signal, black crosses – background; the number of crosses indicates intensity. Scale bar 10 μm. Cx43s – Cx43 antibody from Sigma, Cx43a – Cx43 antibody from Alomone, NB – Neurobiotin.

    Journal: The Journal of comparative neurology

    Article Title: Domain-specific distribution of gap junctions defines cellular coupling to establish a vascular relay in the retina

    doi: 10.1002/cne.24699

    Figure Lengend Snippet: Validation and optimization of Connexin antibodies used in this study. (a–d) Cx37 labeling revealed just a few puncta (green) on blood vessels (blue) in pure PFA fixative (a) and entire outlines of contractile cells in carbodiimide-based fixatives (b and c). Ganglion cells were more visible in PFA fixative (asterisks); the lowest background was in carbodiimide. Pre-absorption of the antibody with a control peptide eliminated the labeling (d). (e–h) Cx40 (green) was associated with outlines of endothelial cells (blue) in all fixative with the less background in carbodiimide (f). Incubation of primary antibody eliminated the labeling (h) with the exception of labeled microglia (arrow). (i–k) Cx43s-positive puncta (green, Sigma antibody) were associated with vasculature in PFA fixative (i); in addition string-like structures along endothelial cell contacts (blue) became visible in carbodiimide fixative (j–k). (l) Omission of the primary antibody eliminated punctate and string-like structures; the microglia was labeled by the secondary antibody (arrow). (m–p) Cx43a produced identical pattern as Cx43s in carbodiimide containing fixatives (n, o). In pure PFA fixative, this antibody labeled well punctate and string-like structures; however the background was significantly higher (m). Incubation of the primary antibodies with control peptide eliminated the specific labeling (p). (q–s) Morphology of DsRed expressing pericytes was best preserved in PFA-containing fixatives (q and s); the DsRed labeling in pericyte processes was lost in pure carbodiimide fixative (q). (t) Summary of fixation procedure. Red crosses indicate signal, black crosses – background; the number of crosses indicates intensity. Scale bar 10 μm. Cx43s – Cx43 antibody from Sigma, Cx43a – Cx43 antibody from Alomone, NB – Neurobiotin.

    Article Snippet: In addition, when the Alomone’s Cx43 antibody was preincubated with the control peptide antigen, the staining in the mouse retina was eliminated ( ).

    Techniques: Labeling, Incubation, Produced, Expressing

    Cx37, Cx40, and Cx43 are expressed in retinal neurons at much lower levels than in the vasculature. (a) Cx37 expression shown in a vertical view, created by rotation of a z-stack from the corresponding retinal whole mount region in (b). (b) Confocal projection of a retinal whole mount at the GCL. Some ganglion cells expressed Cx37 at their cell membrane (asterisks, insert). (c) Blood vessels had higher expression of Cx37 (blue) than ganglion cells (red). (d–e) Cx40 was weakly expressed in numerous nuclei of inner retinal neurons. High expression of Cx40 was detected in blood vessels (f, blue). (g–i) Cx43 was expressed in nuclei (insert) and processes of inner retinal neurons (g, four bright bands in the inner plexiform layer). The expression in neurons was lower than in blood vessels (i). Scale bar 20 μm. ONL – outer nuclear layer, OPL – outer plexiform layer, INL – inner nuclear layer, IPL – inner plexiform layer, GCL – ganglion cell layer.

    Journal: The Journal of comparative neurology

    Article Title: Domain-specific distribution of gap junctions defines cellular coupling to establish a vascular relay in the retina

    doi: 10.1002/cne.24699

    Figure Lengend Snippet: Cx37, Cx40, and Cx43 are expressed in retinal neurons at much lower levels than in the vasculature. (a) Cx37 expression shown in a vertical view, created by rotation of a z-stack from the corresponding retinal whole mount region in (b). (b) Confocal projection of a retinal whole mount at the GCL. Some ganglion cells expressed Cx37 at their cell membrane (asterisks, insert). (c) Blood vessels had higher expression of Cx37 (blue) than ganglion cells (red). (d–e) Cx40 was weakly expressed in numerous nuclei of inner retinal neurons. High expression of Cx40 was detected in blood vessels (f, blue). (g–i) Cx43 was expressed in nuclei (insert) and processes of inner retinal neurons (g, four bright bands in the inner plexiform layer). The expression in neurons was lower than in blood vessels (i). Scale bar 20 μm. ONL – outer nuclear layer, OPL – outer plexiform layer, INL – inner nuclear layer, IPL – inner plexiform layer, GCL – ganglion cell layer.

    Article Snippet: In addition, when the Alomone’s Cx43 antibody was preincubated with the control peptide antigen, the staining in the mouse retina was eliminated ( ).

    Techniques: Expressing

    Cx37 is expressed in all vascular cells. (a) In confocal section through an artery, puncta of cx37 (green) were present on endothelial cells (blue). (b) In the same artery, Cx37 was also co-localized on mural cells (magenta). (c) In the cross section of the artery from (a-b) Cx37 was present on both endothelial and contractile cells as reflected by the intensity profile on the right. (d–e) Cx37 was present on the entire membrane of the mural cells. (f–i) Similar to artery, in a capillary Cx43 was expressed in both endothelial and contractile cells. (j–I) In veins the strongest Cx37 labeling was around pericytes. Cx37 puncta were present on endothelial cells. Scale bar 5 μm.

    Journal: The Journal of comparative neurology

    Article Title: Domain-specific distribution of gap junctions defines cellular coupling to establish a vascular relay in the retina

    doi: 10.1002/cne.24699

    Figure Lengend Snippet: Cx37 is expressed in all vascular cells. (a) In confocal section through an artery, puncta of cx37 (green) were present on endothelial cells (blue). (b) In the same artery, Cx37 was also co-localized on mural cells (magenta). (c) In the cross section of the artery from (a-b) Cx37 was present on both endothelial and contractile cells as reflected by the intensity profile on the right. (d–e) Cx37 was present on the entire membrane of the mural cells. (f–i) Similar to artery, in a capillary Cx43 was expressed in both endothelial and contractile cells. (j–I) In veins the strongest Cx37 labeling was around pericytes. Cx37 puncta were present on endothelial cells. Scale bar 5 μm.

    Article Snippet: In addition, when the Alomone’s Cx43 antibody was preincubated with the control peptide antigen, the staining in the mouse retina was eliminated ( ).

    Techniques: Labeling

    Cx43 promotes vascular coupling between the capillaries and the artery. (a-c) The relay zones were identified based on the staining against smooth muscle actin (SMA). The string-like Cx43-positive structures started at the primary artery and continued uninterrupted beyond SMA-labeling into selected capillaries (arrows). Some capillaries had much weaker or no string-like expression (arrowheads). The strings from the superficial layer in (a) extended into intermediate (b) and deep (c) vascular layer capillaries using the shortest route from the artery. Scale bar 50 μm. (d) In the projection of all layers, string-bearing selected capillaries created a specialized vascular domain – vascular relay - spanning all three layers. The bottom panel shows a vertical view. Scale bar 50 μm. (e) Neurobiotin injected into a pericyte (arrow) in relay area spread throughout the large area. (f) Neurobiotin injected into the area outside the relay was restricted to a few pericytes. (g) Quantification shows that under normal conditions cells are extensively coupled through GJs in vascular relay (data are shown as average ± SD; standard: 3.3 ± 2.1, 15 samples, 8 mice; sensor: 23.7 ± 9.6, 11 samples, 6 mice; T-test, p = 0.00003). (h) Distribution of Cx43-positive strings across vascular layers shows that in the vascular relay region Neurobiotin spread farther into intermediate and deep vascular layers.

    Journal: The Journal of comparative neurology

    Article Title: Domain-specific distribution of gap junctions defines cellular coupling to establish a vascular relay in the retina

    doi: 10.1002/cne.24699

    Figure Lengend Snippet: Cx43 promotes vascular coupling between the capillaries and the artery. (a-c) The relay zones were identified based on the staining against smooth muscle actin (SMA). The string-like Cx43-positive structures started at the primary artery and continued uninterrupted beyond SMA-labeling into selected capillaries (arrows). Some capillaries had much weaker or no string-like expression (arrowheads). The strings from the superficial layer in (a) extended into intermediate (b) and deep (c) vascular layer capillaries using the shortest route from the artery. Scale bar 50 μm. (d) In the projection of all layers, string-bearing selected capillaries created a specialized vascular domain – vascular relay - spanning all three layers. The bottom panel shows a vertical view. Scale bar 50 μm. (e) Neurobiotin injected into a pericyte (arrow) in relay area spread throughout the large area. (f) Neurobiotin injected into the area outside the relay was restricted to a few pericytes. (g) Quantification shows that under normal conditions cells are extensively coupled through GJs in vascular relay (data are shown as average ± SD; standard: 3.3 ± 2.1, 15 samples, 8 mice; sensor: 23.7 ± 9.6, 11 samples, 6 mice; T-test, p = 0.00003). (h) Distribution of Cx43-positive strings across vascular layers shows that in the vascular relay region Neurobiotin spread farther into intermediate and deep vascular layers.

    Article Snippet: In addition, when the Alomone’s Cx43 antibody was preincubated with the control peptide antigen, the staining in the mouse retina was eliminated ( ).

    Techniques: Staining, Labeling, Expressing, Injection, Mouse Assay

    In the arteries and relays gap junctions of endothelial cells strictly colocalize with tight junctions. (a–d) In the artery, Cx40 (green) was strictly colocalized with tight junctions (magenta, arrows) and less precisely with adherens proteins (blue). In a merged image of claudin5 and PECAM (d), the colocalization was not precise. White line highlights the region used for fluorescent intensity profile. Note that claudin5 and Cx40 profiles closely match and differ from the PECAM profile. (e–h) In the vascular relay, Cx43-positive strings were colocalized with tight junctions including strings and small varicosities (arrows). (i–l) In the capillary, Cx43 was not detected along tight junctions. Scale bar 10 μm.

    Journal: The Journal of comparative neurology

    Article Title: Domain-specific distribution of gap junctions defines cellular coupling to establish a vascular relay in the retina

    doi: 10.1002/cne.24699

    Figure Lengend Snippet: In the arteries and relays gap junctions of endothelial cells strictly colocalize with tight junctions. (a–d) In the artery, Cx40 (green) was strictly colocalized with tight junctions (magenta, arrows) and less precisely with adherens proteins (blue). In a merged image of claudin5 and PECAM (d), the colocalization was not precise. White line highlights the region used for fluorescent intensity profile. Note that claudin5 and Cx40 profiles closely match and differ from the PECAM profile. (e–h) In the vascular relay, Cx43-positive strings were colocalized with tight junctions including strings and small varicosities (arrows). (i–l) In the capillary, Cx43 was not detected along tight junctions. Scale bar 10 μm.

    Article Snippet: In addition, when the Alomone’s Cx43 antibody was preincubated with the control peptide antigen, the staining in the mouse retina was eliminated ( ).

    Techniques:

    Histological analysis of cardiac tissue after long-term training. ( A ) Representative images for sedentary, 8 weeks of running and 16 weeks of running tissue from wt (upper row) and PKP2 +/- mice (lower row) are shown for Trichrome-stained ventricular slices. No change of fibrous tissue could be detected in either genotype or in response to long-term training. ( B ) The oil stain in frozen sections from cardiac tissue with oil red did not reveal an increase in fatty replacements. ( C ) We labeled the gap junctions with a Cx43 specific antibody and analyzed the images by confocal microscopy. The connexins were predominantly located at the intercalated discs as seen in the representative images shown. There was no shift in subcellular distribution due to genotype or long-term training observed. The single data points indicate the number of animals analyzed.

    Journal: PLoS ONE

    Article Title: Beneficial effect of voluntary physical exercise in Plakophilin2 transgenic mice

    doi: 10.1371/journal.pone.0252649

    Figure Lengend Snippet: Histological analysis of cardiac tissue after long-term training. ( A ) Representative images for sedentary, 8 weeks of running and 16 weeks of running tissue from wt (upper row) and PKP2 +/- mice (lower row) are shown for Trichrome-stained ventricular slices. No change of fibrous tissue could be detected in either genotype or in response to long-term training. ( B ) The oil stain in frozen sections from cardiac tissue with oil red did not reveal an increase in fatty replacements. ( C ) We labeled the gap junctions with a Cx43 specific antibody and analyzed the images by confocal microscopy. The connexins were predominantly located at the intercalated discs as seen in the representative images shown. There was no shift in subcellular distribution due to genotype or long-term training observed. The single data points indicate the number of animals analyzed.

    Article Snippet: A subset of paraffin sections was used for immunohistochemistry to study the distribution of connexin 43 (Cx43) in the ventricular tissue.

    Techniques: Mouse Assay, Staining, Labeling, Confocal Microscopy