kynurenic acid  (Alomone Labs)


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    Alomone Labs kynurenic acid
    Kynurenic Acid, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs anti twik 1 antibodies
    Ba 2+ at micromolar concentration does not eliminate the inward component of passive conductance in <t>TWIK-1</t> −/− mice. (A) 100 μ M BaCl 2 induced similar Vm depolarization in WT and TWIK-1 −/− astrocytes. Δ V indicates the plateau Vm depolarization at 5 min in BaCl 2 . (B) 100 μ M BaCl 2 induced Δ V was not differed between WT and TWIK-1 −/− astrocytes. (C) The representative whole cell passive currents before and after 5 min 100 μ M BaCl 2 application. I–V relationships were shown in (D) . The Ba 2+ sensitive currents were obtained from sweep subtraction. The Ba 2+ sensitive currents were shown in expanded y-axis in the inset in (D) that showed a moderate inward rectification in both WT, RI = 0.86, and TWIK-1 −/− , RI = 0.87, astrocytes. Data are shown as mean ± SEM. Numbers of experiments are given in the bars.
    Anti Twik 1 Antibodies, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti twik 1 antibodies/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti twik 1 antibodies - by Bioz Stars, 2022-05
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    Ba 2+ at micromolar concentration does not eliminate the inward component of passive conductance in TWIK-1 −/− mice. (A) 100 μ M BaCl 2 induced similar Vm depolarization in WT and TWIK-1 −/− astrocytes. Δ V indicates the plateau Vm depolarization at 5 min in BaCl 2 . (B) 100 μ M BaCl 2 induced Δ V was not differed between WT and TWIK-1 −/− astrocytes. (C) The representative whole cell passive currents before and after 5 min 100 μ M BaCl 2 application. I–V relationships were shown in (D) . The Ba 2+ sensitive currents were obtained from sweep subtraction. The Ba 2+ sensitive currents were shown in expanded y-axis in the inset in (D) that showed a moderate inward rectification in both WT, RI = 0.86, and TWIK-1 −/− , RI = 0.87, astrocytes. Data are shown as mean ± SEM. Numbers of experiments are given in the bars.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: The contribution of TWIK-1 channels to astrocyte K+ current is limited by retention in intracellular compartments

    doi: 10.3389/fncel.2013.00246

    Figure Lengend Snippet: Ba 2+ at micromolar concentration does not eliminate the inward component of passive conductance in TWIK-1 −/− mice. (A) 100 μ M BaCl 2 induced similar Vm depolarization in WT and TWIK-1 −/− astrocytes. Δ V indicates the plateau Vm depolarization at 5 min in BaCl 2 . (B) 100 μ M BaCl 2 induced Δ V was not differed between WT and TWIK-1 −/− astrocytes. (C) The representative whole cell passive currents before and after 5 min 100 μ M BaCl 2 application. I–V relationships were shown in (D) . The Ba 2+ sensitive currents were obtained from sweep subtraction. The Ba 2+ sensitive currents were shown in expanded y-axis in the inset in (D) that showed a moderate inward rectification in both WT, RI = 0.86, and TWIK-1 −/− , RI = 0.87, astrocytes. Data are shown as mean ± SEM. Numbers of experiments are given in the bars.

    Article Snippet: The membranes were incubated with anti-TWIK-1 antibodies (1:2000, Alomone Labs, Jerusalem, Israel) at 4°C overnight.

    Techniques: Concentration Assay, Mouse Assay

    TWIK-1 channels are predominantly located in cytoplasm. (A) Western blot results of TWIK-1 location in cytoplasmic vs . membrane proteins of two mice hippocampus. GFAP (50 kDa) and ATP1α2 (112 kDa) were markers for cytoplasmic and membrane fractions, respectively. Note that TWIK-1 dimer and monomer were mainly located in cytoplasmic fraction in both mice. (B) A bar graph showing the relative ratio of TWIK-1 proteins located in cytoplasmic and membrane fractions. (C) A representative blotting result from the second fractionation method (see Methods). Total protein from hippocampus was fractionated into cytoplasmic proteins (CP), mildly hydrophobic membrane proteins (MHMP), plasma membrane protein (PMP) and lipid raft proteins (LRP). GFAP, ATP1α2 and CAV1 (21 kDa) were markers for CP, PMP and LRP, respectively. TWIK-1 also expressed highly in cytoplasm, relative less in MHMP and very low in PMP fraction. K ir 4.1 (42 kDa), another glial specific K + channel, was detected from both of the MHMP and PMP as anticipated. (D) Quantitation of TWIK-1 subcellular location in the four subcellular regions noted above ( n = 8). (E) A representative blot with the second fractionation method in (C) from kidney proteins. GAPDH and CAV1 were used as cytoplasm and lipid raft markers, respectively. The amount of TWIK-1 channels in PMP fraction was significantly higher in kidney than that of the hippocampus. (F) Comparison of the subcellular location of TWIK-1 between hippocampus ( n = 8) and kidney ( n = 3). The blots shown in (A) , (C) and (E) were all first incubated with anti-TWIK-1 antibody and then re-probed with the rest of primary antibodies sequentially after the original membranes were stripped with stripping buffer. * p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: The contribution of TWIK-1 channels to astrocyte K+ current is limited by retention in intracellular compartments

    doi: 10.3389/fncel.2013.00246

    Figure Lengend Snippet: TWIK-1 channels are predominantly located in cytoplasm. (A) Western blot results of TWIK-1 location in cytoplasmic vs . membrane proteins of two mice hippocampus. GFAP (50 kDa) and ATP1α2 (112 kDa) were markers for cytoplasmic and membrane fractions, respectively. Note that TWIK-1 dimer and monomer were mainly located in cytoplasmic fraction in both mice. (B) A bar graph showing the relative ratio of TWIK-1 proteins located in cytoplasmic and membrane fractions. (C) A representative blotting result from the second fractionation method (see Methods). Total protein from hippocampus was fractionated into cytoplasmic proteins (CP), mildly hydrophobic membrane proteins (MHMP), plasma membrane protein (PMP) and lipid raft proteins (LRP). GFAP, ATP1α2 and CAV1 (21 kDa) were markers for CP, PMP and LRP, respectively. TWIK-1 also expressed highly in cytoplasm, relative less in MHMP and very low in PMP fraction. K ir 4.1 (42 kDa), another glial specific K + channel, was detected from both of the MHMP and PMP as anticipated. (D) Quantitation of TWIK-1 subcellular location in the four subcellular regions noted above ( n = 8). (E) A representative blot with the second fractionation method in (C) from kidney proteins. GAPDH and CAV1 were used as cytoplasm and lipid raft markers, respectively. The amount of TWIK-1 channels in PMP fraction was significantly higher in kidney than that of the hippocampus. (F) Comparison of the subcellular location of TWIK-1 between hippocampus ( n = 8) and kidney ( n = 3). The blots shown in (A) , (C) and (E) were all first incubated with anti-TWIK-1 antibody and then re-probed with the rest of primary antibodies sequentially after the original membranes were stripped with stripping buffer. * p

    Article Snippet: The membranes were incubated with anti-TWIK-1 antibodies (1:2000, Alomone Labs, Jerusalem, Israel) at 4°C overnight.

    Techniques: Western Blot, Mouse Assay, Fractionation, Quantitation Assay, Incubation, Stripping Membranes

    Tissue specific expression of TWIK-1 mRNA and proteins and age-dependent up-regulation of TWIK-1 in hippocampus. (A) RT-PCR analysis of TWIK-1 mRNA expression in various adult mouse tissues. TWIK-1 mRNA expression is high in hippocampus, kidney and lung, but barely detectable from heart, liver and skeletal (Sk.) muscle. GAPDH was measured as internal control. The PCR product size for TWIK-1 and GAPDH gene were 191 bp and 394 bp, respectively. (B) Representative Western blot results analyzed from various tissues noted above. Note that TWIK-1 protein expression followed exactly the same tissue-specific pattern of TWIK-1 mRNA. (C) When treating the hippocampal tissue samples with the reducing agent dithiothreitol (DTT, 20 mM) at different times, the ratio of TWIK-1 dimer versus monomer decreased in a time-dependent manner. (D) A representative blotting result of TWIK-1 proteins from mice hippocampus of different ages as indicated. Both GAPDH (36 kDa) and GFAP (50 kDa) were the loading controls from the same blots. (E) Quantitation of TWIK-1 expression levels from different ages as shown in (D) . Data were normalized to the levels of 3 week (wk.), n = 3. * p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: The contribution of TWIK-1 channels to astrocyte K+ current is limited by retention in intracellular compartments

    doi: 10.3389/fncel.2013.00246

    Figure Lengend Snippet: Tissue specific expression of TWIK-1 mRNA and proteins and age-dependent up-regulation of TWIK-1 in hippocampus. (A) RT-PCR analysis of TWIK-1 mRNA expression in various adult mouse tissues. TWIK-1 mRNA expression is high in hippocampus, kidney and lung, but barely detectable from heart, liver and skeletal (Sk.) muscle. GAPDH was measured as internal control. The PCR product size for TWIK-1 and GAPDH gene were 191 bp and 394 bp, respectively. (B) Representative Western blot results analyzed from various tissues noted above. Note that TWIK-1 protein expression followed exactly the same tissue-specific pattern of TWIK-1 mRNA. (C) When treating the hippocampal tissue samples with the reducing agent dithiothreitol (DTT, 20 mM) at different times, the ratio of TWIK-1 dimer versus monomer decreased in a time-dependent manner. (D) A representative blotting result of TWIK-1 proteins from mice hippocampus of different ages as indicated. Both GAPDH (36 kDa) and GFAP (50 kDa) were the loading controls from the same blots. (E) Quantitation of TWIK-1 expression levels from different ages as shown in (D) . Data were normalized to the levels of 3 week (wk.), n = 3. * p

    Article Snippet: The membranes were incubated with anti-TWIK-1 antibodies (1:2000, Alomone Labs, Jerusalem, Israel) at 4°C overnight.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Western Blot, Mouse Assay, Quantitation Assay

    TWIK-1 deletion does not alter the expression pattern of astrocyte K + channels. (A) qRT-PCR results of the relative quantity of TWIK-1, K ir 4.1, TREK-1, TWIK-2, and TWIK-3 from the total mRNA isolated from mice hippocampus. The expression levels of TWIK-1 mRNAs were reduced to half and hundred percent in TWIK-1 +/− and TWIK-1 −/− mice, respectively. (B) Morphology of freshly isolated astrocyte and neuron from mice hippocampus (DIC). Scale bar: 10 μm. These cells were harvested separately for qRT-PCR analysis. Astrocytes were selected based on their positive SR101 staining (middle up panel), and SR101 staining was completely devoid in isolated pyramidal neurons (middle bottom panel). (C) The expression pattern of TWIK-1, K ir 4.1 and TREK-1 of isolated astrocytes resembled that of the hippocampal tissues (A) . The expression of TWIK-1 and K ir 4.1 appeared to be astrocytic, while TREK-1 was both astrocytic and neuronal. Data were normalized to the expression level of TWIK-1 mRNA in WT. In each genotype group, 30 isolated astrocytes or neurons were harvested from 3 different mice and analyzed separately to obtain a mean ± SEM ( n = 3). ** P

    Journal: Frontiers in Cellular Neuroscience

    Article Title: The contribution of TWIK-1 channels to astrocyte K+ current is limited by retention in intracellular compartments

    doi: 10.3389/fncel.2013.00246

    Figure Lengend Snippet: TWIK-1 deletion does not alter the expression pattern of astrocyte K + channels. (A) qRT-PCR results of the relative quantity of TWIK-1, K ir 4.1, TREK-1, TWIK-2, and TWIK-3 from the total mRNA isolated from mice hippocampus. The expression levels of TWIK-1 mRNAs were reduced to half and hundred percent in TWIK-1 +/− and TWIK-1 −/− mice, respectively. (B) Morphology of freshly isolated astrocyte and neuron from mice hippocampus (DIC). Scale bar: 10 μm. These cells were harvested separately for qRT-PCR analysis. Astrocytes were selected based on their positive SR101 staining (middle up panel), and SR101 staining was completely devoid in isolated pyramidal neurons (middle bottom panel). (C) The expression pattern of TWIK-1, K ir 4.1 and TREK-1 of isolated astrocytes resembled that of the hippocampal tissues (A) . The expression of TWIK-1 and K ir 4.1 appeared to be astrocytic, while TREK-1 was both astrocytic and neuronal. Data were normalized to the expression level of TWIK-1 mRNA in WT. In each genotype group, 30 isolated astrocytes or neurons were harvested from 3 different mice and analyzed separately to obtain a mean ± SEM ( n = 3). ** P

    Article Snippet: The membranes were incubated with anti-TWIK-1 antibodies (1:2000, Alomone Labs, Jerusalem, Israel) at 4°C overnight.

    Techniques: Expressing, Quantitative RT-PCR, Isolation, Mouse Assay, Staining

    TWIK-1 gene deletion alters the membrane properties of astrocytes. (A) Representative whole-cell currents of three genotypes. Cells were held at −80 mV at resting and the membrane currents were induced by command voltages from −180 to +20 mV with 10 mV increment ( inset , command voltages, Vcom). Astrocytes in all genotypes showed a similar linear I–V relationship membrane conductance. (B), (C) TWIK-1 gene deletion hyperpolarized astrocyte membrane potential (Vm, B ), but did not significantly alter the membrane resistance (Rm, C ). (D) To construct I–V curves, the current amplitudes from the dashed vertical lines in (A) were plotted against their corresponding Vcom. The current amplitude at y1 (V 20 mV ) and y2 (V -180 mV ) were used to calculate RI according to equation [1] (see Methods). (E) The RI values shifted from weakly inward rectification in WT to close to linear in TWIK-1 −/− astrocytes in a gene dependent manner. Data are shown as mean ± SEM, * p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: The contribution of TWIK-1 channels to astrocyte K+ current is limited by retention in intracellular compartments

    doi: 10.3389/fncel.2013.00246

    Figure Lengend Snippet: TWIK-1 gene deletion alters the membrane properties of astrocytes. (A) Representative whole-cell currents of three genotypes. Cells were held at −80 mV at resting and the membrane currents were induced by command voltages from −180 to +20 mV with 10 mV increment ( inset , command voltages, Vcom). Astrocytes in all genotypes showed a similar linear I–V relationship membrane conductance. (B), (C) TWIK-1 gene deletion hyperpolarized astrocyte membrane potential (Vm, B ), but did not significantly alter the membrane resistance (Rm, C ). (D) To construct I–V curves, the current amplitudes from the dashed vertical lines in (A) were plotted against their corresponding Vcom. The current amplitude at y1 (V 20 mV ) and y2 (V -180 mV ) were used to calculate RI according to equation [1] (see Methods). (E) The RI values shifted from weakly inward rectification in WT to close to linear in TWIK-1 −/− astrocytes in a gene dependent manner. Data are shown as mean ± SEM, * p

    Article Snippet: The membranes were incubated with anti-TWIK-1 antibodies (1:2000, Alomone Labs, Jerusalem, Israel) at 4°C overnight.

    Techniques: Construct

    Effects of quinine on astrocyte membrane potential and passive conductance in WT and TWIK-1 −/− mice. (A) 0.4 mM quinine induced comparable amplitude of Vm depolarization in both WT and TWIK-1 −/− astrocyte. ΔV indicates the plateau amplitude of Vm depolarization at 8 min of quinine application. (B) Summary of ΔV from both genotypes. (C) The representative whole-cell passive currents before and after 8 min 0.4 mM quinine application. The Vcom protocol shown in Figure 3A was also used here for membrane current induction. I–V relationships were shown in (D) . The quinine sensitive currents were obtained from sweep subtraction, which were linear I–V relationship in both genotypes. Data are shown as mean ± SEM.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: The contribution of TWIK-1 channels to astrocyte K+ current is limited by retention in intracellular compartments

    doi: 10.3389/fncel.2013.00246

    Figure Lengend Snippet: Effects of quinine on astrocyte membrane potential and passive conductance in WT and TWIK-1 −/− mice. (A) 0.4 mM quinine induced comparable amplitude of Vm depolarization in both WT and TWIK-1 −/− astrocyte. ΔV indicates the plateau amplitude of Vm depolarization at 8 min of quinine application. (B) Summary of ΔV from both genotypes. (C) The representative whole-cell passive currents before and after 8 min 0.4 mM quinine application. The Vcom protocol shown in Figure 3A was also used here for membrane current induction. I–V relationships were shown in (D) . The quinine sensitive currents were obtained from sweep subtraction, which were linear I–V relationship in both genotypes. Data are shown as mean ± SEM.

    Article Snippet: The membranes were incubated with anti-TWIK-1 antibodies (1:2000, Alomone Labs, Jerusalem, Israel) at 4°C overnight.

    Techniques: Mouse Assay

    TWIK-1 gene deletion decreases the relative Cs + to K + permeability of passive conductance. (A) Astrocyte membrane potential (Vm) approaches to Cs + equilibrium potential ( E Cs ) in Cs + -based pipette and bath solutions. Switch extracellular 3.5 mM K + to 3.5 mM Cs + resulted in a rapid negative Vm shift in all three genotypes with a net negative shift of around −10 mV at the plateau in 2–3 min. The plateau Vm values at the time point indicated by “*” were summarized in (B) . The Cs + established Vm showed a TWIK-1 gene dependent positive shift and the difference between WT and TWIK-1 −/− mice was statistically significant. (C) Representative whole-cell membrane potential recording traces, with bath solutions switched from normal aCSF containing 3.5 mM K + to solutions containing, in sequence, 70 mM K + and 70 mM Cs + . E Cs and E K are the equilibrium potential of Cs + and K + at 70 mM, respectively. (D) The relative Cs + to K + permeability ( P Cs / P K ) values for three genotypes; the P Cs / P K of TWIK-1 −/− mice was significantly lower than that of the TWIK-1. Data are shown as mean ± SEM. Numbers of experiments are given within the bars. * p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: The contribution of TWIK-1 channels to astrocyte K+ current is limited by retention in intracellular compartments

    doi: 10.3389/fncel.2013.00246

    Figure Lengend Snippet: TWIK-1 gene deletion decreases the relative Cs + to K + permeability of passive conductance. (A) Astrocyte membrane potential (Vm) approaches to Cs + equilibrium potential ( E Cs ) in Cs + -based pipette and bath solutions. Switch extracellular 3.5 mM K + to 3.5 mM Cs + resulted in a rapid negative Vm shift in all three genotypes with a net negative shift of around −10 mV at the plateau in 2–3 min. The plateau Vm values at the time point indicated by “*” were summarized in (B) . The Cs + established Vm showed a TWIK-1 gene dependent positive shift and the difference between WT and TWIK-1 −/− mice was statistically significant. (C) Representative whole-cell membrane potential recording traces, with bath solutions switched from normal aCSF containing 3.5 mM K + to solutions containing, in sequence, 70 mM K + and 70 mM Cs + . E Cs and E K are the equilibrium potential of Cs + and K + at 70 mM, respectively. (D) The relative Cs + to K + permeability ( P Cs / P K ) values for three genotypes; the P Cs / P K of TWIK-1 −/− mice was significantly lower than that of the TWIK-1. Data are shown as mean ± SEM. Numbers of experiments are given within the bars. * p

    Article Snippet: The membranes were incubated with anti-TWIK-1 antibodies (1:2000, Alomone Labs, Jerusalem, Israel) at 4°C overnight.

    Techniques: Permeability, Transferring, Mouse Assay, Sequencing

    TWIK-1 gene knockout mice. (A) Schematic illustration of the targeted exon 2 deletion in TWIK-1 gene. Exon 2 encodes the two ion conducting pores (P1, P2), and the transmembrane domain 2 and 3 (M2, M3) (all in shadow). (B) RT-PCR amplification targeted on the axon 2 of TWIK-1 gene was performed with the mRNAs extracted from hippocampus of WT, heterozygote (TWIK-1 +/− ) and TWIK-1 −/− mice, that yielded the anticipated products as indicated. (C) Sequencing analysis from the PCR products from (B) confirmed an anticipated deletion of exon 2 in TWIK-1 −/− mouse, which encodes 132 amino acids in the position 119–250 of TWIK-1 channel.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: The contribution of TWIK-1 channels to astrocyte K+ current is limited by retention in intracellular compartments

    doi: 10.3389/fncel.2013.00246

    Figure Lengend Snippet: TWIK-1 gene knockout mice. (A) Schematic illustration of the targeted exon 2 deletion in TWIK-1 gene. Exon 2 encodes the two ion conducting pores (P1, P2), and the transmembrane domain 2 and 3 (M2, M3) (all in shadow). (B) RT-PCR amplification targeted on the axon 2 of TWIK-1 gene was performed with the mRNAs extracted from hippocampus of WT, heterozygote (TWIK-1 +/− ) and TWIK-1 −/− mice, that yielded the anticipated products as indicated. (C) Sequencing analysis from the PCR products from (B) confirmed an anticipated deletion of exon 2 in TWIK-1 −/− mouse, which encodes 132 amino acids in the position 119–250 of TWIK-1 channel.

    Article Snippet: The membranes were incubated with anti-TWIK-1 antibodies (1:2000, Alomone Labs, Jerusalem, Israel) at 4°C overnight.

    Techniques: Gene Knockout, Mouse Assay, Reverse Transcription Polymerase Chain Reaction, Amplification, Sequencing, Polymerase Chain Reaction

    Enhanced potentiation of mGluR3-mediated NH 4 + uptake in the absence of Na + -K + ATPase activity. a To eliminate Na + -K + ATPase (NKA) activity, NaCl in the electrode solution (14 mM) and aCSF (125 mM) was substituted by equimolar LiCl, and the perfusate was switched from normal to high LiCl-containing aCSF (Li-aCSF) after whole-cell recording was established. These conditions depolarized V M by ~10 mV resulting from inhibition of electrogenic NKA activity. NH 4 Cl (5 mM, 10 min) was applied 9~10 min after NKA inhibition. Right panel , the NH 4 Cl response outlined in the dashed rectangle in left panel is displayed in an enlarged scale ( green ) and superimposed with a recording with intact NKA activity ( black ). The differences in NH 4 Cl response under these two conditions are summarized in b ; elimination of NKA activity resulted in a (1) progressive increase in NH 4 + -induced V M depolarization ( left , quantified as “time to peak”), (2) an average of 22.0 % increase in peak amplitude ( middle ), and (3) a total abolishment of V M undershot upon withdrawal of NH 4 Cl ( right ). c Representative recording showing that mGluR3 activation induced potentiation was more pronounced in the absence of NKA activity. d mGluR3-induced potentiation of NH 4 + response remained when NKA is inactive in WT but not in TWIK-1 KO astrocytes. Data are presented as mean±SEM from the indicated number of experiments. ** P

    Journal: Molecular neurobiology

    Article Title: mGluR3 Activation Recruits Cytoplasmic TWIK-1 Channels to Membrane that Enhances Ammonium Uptake in Hippocampal Astrocytes

    doi: 10.1007/s12035-015-9496-4

    Figure Lengend Snippet: Enhanced potentiation of mGluR3-mediated NH 4 + uptake in the absence of Na + -K + ATPase activity. a To eliminate Na + -K + ATPase (NKA) activity, NaCl in the electrode solution (14 mM) and aCSF (125 mM) was substituted by equimolar LiCl, and the perfusate was switched from normal to high LiCl-containing aCSF (Li-aCSF) after whole-cell recording was established. These conditions depolarized V M by ~10 mV resulting from inhibition of electrogenic NKA activity. NH 4 Cl (5 mM, 10 min) was applied 9~10 min after NKA inhibition. Right panel , the NH 4 Cl response outlined in the dashed rectangle in left panel is displayed in an enlarged scale ( green ) and superimposed with a recording with intact NKA activity ( black ). The differences in NH 4 Cl response under these two conditions are summarized in b ; elimination of NKA activity resulted in a (1) progressive increase in NH 4 + -induced V M depolarization ( left , quantified as “time to peak”), (2) an average of 22.0 % increase in peak amplitude ( middle ), and (3) a total abolishment of V M undershot upon withdrawal of NH 4 Cl ( right ). c Representative recording showing that mGluR3 activation induced potentiation was more pronounced in the absence of NKA activity. d mGluR3-induced potentiation of NH 4 + response remained when NKA is inactive in WT but not in TWIK-1 KO astrocytes. Data are presented as mean±SEM from the indicated number of experiments. ** P

    Article Snippet: They were rabbit anti-TWIK-1 (1:400, Alomone Labs, Jerusalem, Israel), guinea pig anti-GLT1 (1:1000, Millipore, Billerica, MA), goat anti-GFAP (1:1000, Abcam, Cambridge, MA), or rat anti-Rab11 (1: 500, Abcam, Cambridge, MA).

    Techniques: Activity Assay, Inhibition, Activation Assay

    Cellular identification of GFP-expressing cells of the DG, LEC, and Cb in P56 of TWIK-1 BAC-GFP Tg mice. ( A ) Overview of GFP expression in DG, LEC, and CB of TWIK-1 BAC-GFP Tg mice. Scale bar, 200 μm. ( B ) Representative co-immunofluorescence images with GFP, NeuN, and GAD67 antibodies. Scale bar, 200 μm. ( C ) Quantification bar graph of the cell type of GFP-positive cells in each brain area from B. Quantification was analyzed by the percentage of each cell type from all GFP-positive cells. Raw data are listed in Supplementary Materials Table S1 . Data are presented as the Mean ± SEM.

    Journal: Cells

    Article Title: TWIK-1 BAC-GFP Transgenic Mice, an Animal Model for TWIK-1 Expression

    doi: 10.3390/cells10102751

    Figure Lengend Snippet: Cellular identification of GFP-expressing cells of the DG, LEC, and Cb in P56 of TWIK-1 BAC-GFP Tg mice. ( A ) Overview of GFP expression in DG, LEC, and CB of TWIK-1 BAC-GFP Tg mice. Scale bar, 200 μm. ( B ) Representative co-immunofluorescence images with GFP, NeuN, and GAD67 antibodies. Scale bar, 200 μm. ( C ) Quantification bar graph of the cell type of GFP-positive cells in each brain area from B. Quantification was analyzed by the percentage of each cell type from all GFP-positive cells. Raw data are listed in Supplementary Materials Table S1 . Data are presented as the Mean ± SEM.

    Article Snippet: The following antibodies were used: chicken anti-GFP (Abcam, Cat#; ab136970, 1:300); rabbit anti-TWIK-1 (Alomone labs, Cat#; APC-10, 1:200); mouse anti-NeuN (Abcam, Cat#; ab104224, 1:100); rabbit anti-GAD67 (GeneTex, Cat#; GTX113190, 1:300); guinea pig anti-doublecortin (Millipore, Cat#; AB2253, 1:100); rabbit anti-calbindin (Swant, Cat#; CB38, 1:2000); rat anti-Ki67 (Invitrogen, Cat#; 14-5698-85, 1:200); rat anti-GFAP (Invitrogen, Cat#; 13-0300, 1:500); rabbit anti-NG2 (Millipore, Cat#; AB5320, 1:300); and Alexa Fluor 488-, 594-, and 647-conjugated secondary antibodies (Jackson ImmunoResearch, 1:300).

    Techniques: Expressing, BAC Assay, Mouse Assay, Immunofluorescence

    Generation of TWIK-1 BAC-GFP Tg mice. ( A ) Schematic design of the modified Kcnk1 BAC vector. GFP coding sequence was inserted just before the start codon of Kcnk1 . ( B ) Genotype PCR results for the designed primer pair in A. ( C ) Representative macroscopic GFP fluorescent images in the adult brain (P56 age) from TWIK-1 BAC-GFP Tg mice. Upper left numbers indicate the relative position of the brain slice from bregma. Strong GFP expression was identified in DG, LEC, and Cb (yellow dashed arrow), Scale bar, 1000 μm. ( D ) Representative co-immunofluorescence images with GFP and TWIK-1 in the DG. Scale bar, 200 μm. ( E ) Enlarged inset from D. Scale bar, 50 μm. ( F ) Representative co-immunofluorescence images with GFP and TWIK-1 in the LEC. Scale bar, 200 μm. ( G ) Enlarged inset from F. Scale bar, 50 μm. ( H ) Representative co-immunofluorescence images with GFP and TWIK-1 in the Cb. Scale bar, 200 μm. ( I ) Enlarged inset from H. Scale bar, 50 μm.

    Journal: Cells

    Article Title: TWIK-1 BAC-GFP Transgenic Mice, an Animal Model for TWIK-1 Expression

    doi: 10.3390/cells10102751

    Figure Lengend Snippet: Generation of TWIK-1 BAC-GFP Tg mice. ( A ) Schematic design of the modified Kcnk1 BAC vector. GFP coding sequence was inserted just before the start codon of Kcnk1 . ( B ) Genotype PCR results for the designed primer pair in A. ( C ) Representative macroscopic GFP fluorescent images in the adult brain (P56 age) from TWIK-1 BAC-GFP Tg mice. Upper left numbers indicate the relative position of the brain slice from bregma. Strong GFP expression was identified in DG, LEC, and Cb (yellow dashed arrow), Scale bar, 1000 μm. ( D ) Representative co-immunofluorescence images with GFP and TWIK-1 in the DG. Scale bar, 200 μm. ( E ) Enlarged inset from D. Scale bar, 50 μm. ( F ) Representative co-immunofluorescence images with GFP and TWIK-1 in the LEC. Scale bar, 200 μm. ( G ) Enlarged inset from F. Scale bar, 50 μm. ( H ) Representative co-immunofluorescence images with GFP and TWIK-1 in the Cb. Scale bar, 200 μm. ( I ) Enlarged inset from H. Scale bar, 50 μm.

    Article Snippet: The following antibodies were used: chicken anti-GFP (Abcam, Cat#; ab136970, 1:300); rabbit anti-TWIK-1 (Alomone labs, Cat#; APC-10, 1:200); mouse anti-NeuN (Abcam, Cat#; ab104224, 1:100); rabbit anti-GAD67 (GeneTex, Cat#; GTX113190, 1:300); guinea pig anti-doublecortin (Millipore, Cat#; AB2253, 1:100); rabbit anti-calbindin (Swant, Cat#; CB38, 1:2000); rat anti-Ki67 (Invitrogen, Cat#; 14-5698-85, 1:200); rat anti-GFAP (Invitrogen, Cat#; 13-0300, 1:500); rabbit anti-NG2 (Millipore, Cat#; AB5320, 1:300); and Alexa Fluor 488-, 594-, and 647-conjugated secondary antibodies (Jackson ImmunoResearch, 1:300).

    Techniques: BAC Assay, Mouse Assay, Modification, Plasmid Preparation, Sequencing, Polymerase Chain Reaction, Slice Preparation, Expressing, Immunofluorescence

    Glial expression of TWIK-1 in TWIK-1 BAC-GFP Tg mice. ( A ) Representative co-immunofluorescence images with TWIK-1 and GFAP. Scale bar, 200 μm. ( B ) Enlarged inset from A. Yellow arrow indicates double immunoreactive cells with TWIK-1 and GFAP. Scale bar, 50 μm. ( C ) Representative co-immunofluorescence images with GFP and GFAP. Scale bar, 200 μm. ( D ) Enlarged inset from C. Yellow arrow indicates double immunoreactive cells with GFP and GFAP. Scale bar, 50 μm. ( E ) Representative co-immunofluorescence images with GFP and Iba1. Scale bar, 200 μm. ( F ) Enlarged inset from E. There are no Iba1-positive glial-like GFP-expressing cells (White arrow). Scale bar, 50 μm. ( G ) Representative co-immunofluorescence images with GFP and NG2. Scale bar, 200 μm. ( H ) Enlarged inset from G. There are no NG2-positive glial-like GFP-expressing cells (White arrow). Scale bar, 50 μm.

    Journal: Cells

    Article Title: TWIK-1 BAC-GFP Transgenic Mice, an Animal Model for TWIK-1 Expression

    doi: 10.3390/cells10102751

    Figure Lengend Snippet: Glial expression of TWIK-1 in TWIK-1 BAC-GFP Tg mice. ( A ) Representative co-immunofluorescence images with TWIK-1 and GFAP. Scale bar, 200 μm. ( B ) Enlarged inset from A. Yellow arrow indicates double immunoreactive cells with TWIK-1 and GFAP. Scale bar, 50 μm. ( C ) Representative co-immunofluorescence images with GFP and GFAP. Scale bar, 200 μm. ( D ) Enlarged inset from C. Yellow arrow indicates double immunoreactive cells with GFP and GFAP. Scale bar, 50 μm. ( E ) Representative co-immunofluorescence images with GFP and Iba1. Scale bar, 200 μm. ( F ) Enlarged inset from E. There are no Iba1-positive glial-like GFP-expressing cells (White arrow). Scale bar, 50 μm. ( G ) Representative co-immunofluorescence images with GFP and NG2. Scale bar, 200 μm. ( H ) Enlarged inset from G. There are no NG2-positive glial-like GFP-expressing cells (White arrow). Scale bar, 50 μm.

    Article Snippet: The following antibodies were used: chicken anti-GFP (Abcam, Cat#; ab136970, 1:300); rabbit anti-TWIK-1 (Alomone labs, Cat#; APC-10, 1:200); mouse anti-NeuN (Abcam, Cat#; ab104224, 1:100); rabbit anti-GAD67 (GeneTex, Cat#; GTX113190, 1:300); guinea pig anti-doublecortin (Millipore, Cat#; AB2253, 1:100); rabbit anti-calbindin (Swant, Cat#; CB38, 1:2000); rat anti-Ki67 (Invitrogen, Cat#; 14-5698-85, 1:200); rat anti-GFAP (Invitrogen, Cat#; 13-0300, 1:500); rabbit anti-NG2 (Millipore, Cat#; AB5320, 1:300); and Alexa Fluor 488-, 594-, and 647-conjugated secondary antibodies (Jackson ImmunoResearch, 1:300).

    Techniques: Expressing, BAC Assay, Mouse Assay, Immunofluorescence

    High TWIK-1 expression in immature neurons of the DG in TWIK-1 BAC-GFP Tg mice. ( A ) Representative co-immunofluorescence images with GFP, DCX, CB, and Ki67 in P56 of DG. Scale bar, 200 μm. ( B ) Enlarged inset from A. Most of strong GFP-expressing cells co-labeled with DCX (yellow arrow), but not with CB and Ki67 (white arrow). Scale bar, 10 μm. ( C , D ) Quantification of the cell type of strong GFP-expressing cells. Raw data are listed in Supplementary Materials Table S1 . Data are presented as the Mean ± SEM.

    Journal: Cells

    Article Title: TWIK-1 BAC-GFP Transgenic Mice, an Animal Model for TWIK-1 Expression

    doi: 10.3390/cells10102751

    Figure Lengend Snippet: High TWIK-1 expression in immature neurons of the DG in TWIK-1 BAC-GFP Tg mice. ( A ) Representative co-immunofluorescence images with GFP, DCX, CB, and Ki67 in P56 of DG. Scale bar, 200 μm. ( B ) Enlarged inset from A. Most of strong GFP-expressing cells co-labeled with DCX (yellow arrow), but not with CB and Ki67 (white arrow). Scale bar, 10 μm. ( C , D ) Quantification of the cell type of strong GFP-expressing cells. Raw data are listed in Supplementary Materials Table S1 . Data are presented as the Mean ± SEM.

    Article Snippet: The following antibodies were used: chicken anti-GFP (Abcam, Cat#; ab136970, 1:300); rabbit anti-TWIK-1 (Alomone labs, Cat#; APC-10, 1:200); mouse anti-NeuN (Abcam, Cat#; ab104224, 1:100); rabbit anti-GAD67 (GeneTex, Cat#; GTX113190, 1:300); guinea pig anti-doublecortin (Millipore, Cat#; AB2253, 1:100); rabbit anti-calbindin (Swant, Cat#; CB38, 1:2000); rat anti-Ki67 (Invitrogen, Cat#; 14-5698-85, 1:200); rat anti-GFAP (Invitrogen, Cat#; 13-0300, 1:500); rabbit anti-NG2 (Millipore, Cat#; AB5320, 1:300); and Alexa Fluor 488-, 594-, and 647-conjugated secondary antibodies (Jackson ImmunoResearch, 1:300).

    Techniques: Expressing, BAC Assay, Mouse Assay, Immunofluorescence, Labeling

    TWIK-1 BAC-GFP Tg mice represent kainic acid (KA)-induced increase of TWIK-1 expression. ( A ) Representative GFP expression in a brain slice from saline or KA-treated TWIK-1 BAC-GFP Tg mice. Scale bar, 1000 μm. ( B ) Representative GFP expression in the DG from saline or KA-treated TWIK-1 BAC-GFP Tg mice. Scale bar, 200 μm. Enlarged inset from the experiment. Scale bar, 50 μm. ( C ) Quantification of relative mean GFP intensity of the granule cell layer. ( D ) Representative Kcnk1 mRNA fluorescence in situ hybridization (FISH) images of DG from saline- or KA-treated mice. Scale bar, 100 μm. ( E ) Quantification bar graph of Kcnk1 mRNA spot density in the granule cell layer from saline- or KA-treated mice. ( F ) Representative TWIK-1 immunofluorescence images of DG from saline- or KA-treated mice. Scale bar, 200 μm. ( G ) Quantification bar graph of the relative mean TWIK-1 immunofluorescence in the granule cell layer of saline- or KA-treated mice. Raw data are listed in Supplementary Materials Table S1 . **** p

    Journal: Cells

    Article Title: TWIK-1 BAC-GFP Transgenic Mice, an Animal Model for TWIK-1 Expression

    doi: 10.3390/cells10102751

    Figure Lengend Snippet: TWIK-1 BAC-GFP Tg mice represent kainic acid (KA)-induced increase of TWIK-1 expression. ( A ) Representative GFP expression in a brain slice from saline or KA-treated TWIK-1 BAC-GFP Tg mice. Scale bar, 1000 μm. ( B ) Representative GFP expression in the DG from saline or KA-treated TWIK-1 BAC-GFP Tg mice. Scale bar, 200 μm. Enlarged inset from the experiment. Scale bar, 50 μm. ( C ) Quantification of relative mean GFP intensity of the granule cell layer. ( D ) Representative Kcnk1 mRNA fluorescence in situ hybridization (FISH) images of DG from saline- or KA-treated mice. Scale bar, 100 μm. ( E ) Quantification bar graph of Kcnk1 mRNA spot density in the granule cell layer from saline- or KA-treated mice. ( F ) Representative TWIK-1 immunofluorescence images of DG from saline- or KA-treated mice. Scale bar, 200 μm. ( G ) Quantification bar graph of the relative mean TWIK-1 immunofluorescence in the granule cell layer of saline- or KA-treated mice. Raw data are listed in Supplementary Materials Table S1 . **** p

    Article Snippet: The following antibodies were used: chicken anti-GFP (Abcam, Cat#; ab136970, 1:300); rabbit anti-TWIK-1 (Alomone labs, Cat#; APC-10, 1:200); mouse anti-NeuN (Abcam, Cat#; ab104224, 1:100); rabbit anti-GAD67 (GeneTex, Cat#; GTX113190, 1:300); guinea pig anti-doublecortin (Millipore, Cat#; AB2253, 1:100); rabbit anti-calbindin (Swant, Cat#; CB38, 1:2000); rat anti-Ki67 (Invitrogen, Cat#; 14-5698-85, 1:200); rat anti-GFAP (Invitrogen, Cat#; 13-0300, 1:500); rabbit anti-NG2 (Millipore, Cat#; AB5320, 1:300); and Alexa Fluor 488-, 594-, and 647-conjugated secondary antibodies (Jackson ImmunoResearch, 1:300).

    Techniques: BAC Assay, Mouse Assay, Expressing, Slice Preparation, Fluorescence, In Situ Hybridization, Fluorescence In Situ Hybridization, Immunofluorescence