ca2  (Thermo Fisher)


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    Name:
    CA2 Recombinant Human Protein
    Description:
    Carbonic anhydrase II CA2 recombinant human protein is supplied as a lyophilized powder It is suitable for use in analysis of protein structure In general recombinant proteins can also be used as an immunogen as a protein standard or in cell biology research applications This recombinant protein was expressed from a DNA sequence encoding the human CA2 NP 000058 1 Met 1 Lys 260 fused to the polyhistidinetag at the C terminus Activity Measured by its esterase activity The specific activity is 500 pmoles min µg Formulation Lyophilized in 20 mM Tris 0 5 M NaCl pH 8 0 5 Mannitol 5 Trehalose 0 02 Tween 80 Reconstitution Dissolve the protein in sterile double distilled water to a concentration of 0 2 mg ml or lower It is recommended that the protein be aliquoted and be used as soon as possible Store aliquots under sterile conditions at 20°C Avoid repeated freeze thaw cycles Expiration Date Expires one year from date of receipt when stored as instructed This protein is manufactured by Sino Biological Inc
    Catalog Number:
    10478h08e50
    Price:
    None
    Applications:
    Protein Biology
    Category:
    Proteins Enzymes Peptides
    Buy from Supplier


    Structured Review

    Thermo Fisher ca2
    Lack of homology among Slo3 sequences in the ligand-sensing cytosolic domain. ( A ) Alignments of human Slo1 and Slo3 from various species are shown for the membrane-associated, pore-forming part of the channels, indicating the relatively high extent of conservation through this part of the Slo3 protein. The Slo1 N-terminus is omitted to minimize effects of S0–S1 linker gaps on the alignment. Slo1 numbering starts from amino acids MDAL. Tick marks below each segment of residues counts every 10 residues in human Slo3. ( B ) Alignments of human Slo1 and Slo3 from various species are shown for the cytosolic gating ring domain beginning with the conserved sequence at the beginning of the first RCK domain. Blue highlights residue differences between human Slo1 and human Slo3. Yellow highlights differences of Slo3 of various species to human Slo3. Alignments were generated by Clustal 1.2.0 and minor adjustments were made based on structural considerations ( Leonetti et al., 2012 ). Above the residues, the correspondence of particular amino acid segments to structurally defined α-helical and β-strand segments is shown based on Leonetti et al. (2012) . In red, residues or segments identified in Slo1 or Slo3 isoforms which are implicated in ligand-sensing or species-specific functional differences are highlighted. Although extensive information is available regarding loci important in ligand-sensing in Slo1, such information for Slo3 remains lacking. Numbers identify the following: 1, the sequence of residues termed the Ca 2+ bowl ( Schreiber and Salkoff, 1997 ), for which there is good correspondence of mutations affecting <t>Ca2+-dependent</t> function ( Bao et al., 2004 ) and coordination of density in a crystal structure ( Yuan et al., 2012 ); 2, the D367 residue implicated in the role of the RCK1 domain in Ca 2+ -dependent activation ( Xia et al., 2002 ) which is clearly distinct from Ca 2+ -bowl dependent activation ( Zeng et al., 2005 ); 3, the M513 residue, which also affects Ca 2+− dependent activation involving the RCK1 domain ( Bao et al., 2002 ), but probably is not involved in ligand coordination; 4, residues E374 and E399 which have been implicated in low affinity effects of divalent cations, specifically Mg 2+ ( Shi et al., 2002 ; Xia et al., 2002 ; Yang et al., 2006 ); 5, residue E535 which may also be involved in Ca 2+ coordination in RCK1 ( Zhang et al., 2010 ); 6, residues H365 and H394, which have been implicated in proton-dependent activation of Slo1 and also influence Ca 2+ -dependent activation when protonated ( Hou et al., 2008 ); 7, H417 and segment 368–475, which influence pH-sensing in mouse Slo3 (Zhang et al., 2006); 8, segment 495–515 in bovine Slo3 which accounts for part of the different in functional properties between mouse Slo3 and bovine Slo3 ( Santi et al., 2009 ). Illustrated sequences and accession numbers include: HsSlo1 ( Homo sapiens ), NP_001154824, Gene ID 3778; HsSlo3 ( Homo sapiens ), NP_001027006, Gene ID 157855; MmSlo3 ( Mus musculus ), NP_032458, Gene ID 16532; RnSlo3 ( Rattus norvegicus ), XP_006253398, Gene ID 680912; TcSlo3 (Tupaia chinensis, Chinese tree shrew), XP_006171561, Gene ID 102493286; CcSlo3 ( Condylura cristata , star-nosed mole), XP_004682520, Gene ID 101620543; CfSlo3 (Canis lupus familiaris), XP_539971, Gene ID 482856; BtSlo3 (Bos taurus), NP_001156721, Gene ID 524144; OaSlo3 ( Ovis aries , sheep), XP_004021821, Gene ID 10110209. DOI: http://dx.doi.org/10.7554/eLife.01438.019
    Carbonic anhydrase II CA2 recombinant human protein is supplied as a lyophilized powder It is suitable for use in analysis of protein structure In general recombinant proteins can also be used as an immunogen as a protein standard or in cell biology research applications This recombinant protein was expressed from a DNA sequence encoding the human CA2 NP 000058 1 Met 1 Lys 260 fused to the polyhistidinetag at the C terminus Activity Measured by its esterase activity The specific activity is 500 pmoles min µg Formulation Lyophilized in 20 mM Tris 0 5 M NaCl pH 8 0 5 Mannitol 5 Trehalose 0 02 Tween 80 Reconstitution Dissolve the protein in sterile double distilled water to a concentration of 0 2 mg ml or lower It is recommended that the protein be aliquoted and be used as soon as possible Store aliquots under sterile conditions at 20°C Avoid repeated freeze thaw cycles Expiration Date Expires one year from date of receipt when stored as instructed This protein is manufactured by Sino Biological Inc
    https://www.bioz.com/result/ca2/product/Thermo Fisher
    Average 99 stars, based on 277 article reviews
    Price from $9.99 to $1999.99
    ca2 - by Bioz Stars, 2020-11
    99/100 stars

    Images

    1) Product Images from "The Ca2+-activated K+ current of human sperm is mediated by Slo3"

    Article Title: The Ca2+-activated K+ current of human sperm is mediated by Slo3

    Journal: eLife

    doi: 10.7554/eLife.01438

    Lack of homology among Slo3 sequences in the ligand-sensing cytosolic domain. ( A ) Alignments of human Slo1 and Slo3 from various species are shown for the membrane-associated, pore-forming part of the channels, indicating the relatively high extent of conservation through this part of the Slo3 protein. The Slo1 N-terminus is omitted to minimize effects of S0–S1 linker gaps on the alignment. Slo1 numbering starts from amino acids MDAL. Tick marks below each segment of residues counts every 10 residues in human Slo3. ( B ) Alignments of human Slo1 and Slo3 from various species are shown for the cytosolic gating ring domain beginning with the conserved sequence at the beginning of the first RCK domain. Blue highlights residue differences between human Slo1 and human Slo3. Yellow highlights differences of Slo3 of various species to human Slo3. Alignments were generated by Clustal 1.2.0 and minor adjustments were made based on structural considerations ( Leonetti et al., 2012 ). Above the residues, the correspondence of particular amino acid segments to structurally defined α-helical and β-strand segments is shown based on Leonetti et al. (2012) . In red, residues or segments identified in Slo1 or Slo3 isoforms which are implicated in ligand-sensing or species-specific functional differences are highlighted. Although extensive information is available regarding loci important in ligand-sensing in Slo1, such information for Slo3 remains lacking. Numbers identify the following: 1, the sequence of residues termed the Ca 2+ bowl ( Schreiber and Salkoff, 1997 ), for which there is good correspondence of mutations affecting Ca2+-dependent function ( Bao et al., 2004 ) and coordination of density in a crystal structure ( Yuan et al., 2012 ); 2, the D367 residue implicated in the role of the RCK1 domain in Ca 2+ -dependent activation ( Xia et al., 2002 ) which is clearly distinct from Ca 2+ -bowl dependent activation ( Zeng et al., 2005 ); 3, the M513 residue, which also affects Ca 2+− dependent activation involving the RCK1 domain ( Bao et al., 2002 ), but probably is not involved in ligand coordination; 4, residues E374 and E399 which have been implicated in low affinity effects of divalent cations, specifically Mg 2+ ( Shi et al., 2002 ; Xia et al., 2002 ; Yang et al., 2006 ); 5, residue E535 which may also be involved in Ca 2+ coordination in RCK1 ( Zhang et al., 2010 ); 6, residues H365 and H394, which have been implicated in proton-dependent activation of Slo1 and also influence Ca 2+ -dependent activation when protonated ( Hou et al., 2008 ); 7, H417 and segment 368–475, which influence pH-sensing in mouse Slo3 (Zhang et al., 2006); 8, segment 495–515 in bovine Slo3 which accounts for part of the different in functional properties between mouse Slo3 and bovine Slo3 ( Santi et al., 2009 ). Illustrated sequences and accession numbers include: HsSlo1 ( Homo sapiens ), NP_001154824, Gene ID 3778; HsSlo3 ( Homo sapiens ), NP_001027006, Gene ID 157855; MmSlo3 ( Mus musculus ), NP_032458, Gene ID 16532; RnSlo3 ( Rattus norvegicus ), XP_006253398, Gene ID 680912; TcSlo3 (Tupaia chinensis, Chinese tree shrew), XP_006171561, Gene ID 102493286; CcSlo3 ( Condylura cristata , star-nosed mole), XP_004682520, Gene ID 101620543; CfSlo3 (Canis lupus familiaris), XP_539971, Gene ID 482856; BtSlo3 (Bos taurus), NP_001156721, Gene ID 524144; OaSlo3 ( Ovis aries , sheep), XP_004021821, Gene ID 10110209. DOI: http://dx.doi.org/10.7554/eLife.01438.019
    Figure Legend Snippet: Lack of homology among Slo3 sequences in the ligand-sensing cytosolic domain. ( A ) Alignments of human Slo1 and Slo3 from various species are shown for the membrane-associated, pore-forming part of the channels, indicating the relatively high extent of conservation through this part of the Slo3 protein. The Slo1 N-terminus is omitted to minimize effects of S0–S1 linker gaps on the alignment. Slo1 numbering starts from amino acids MDAL. Tick marks below each segment of residues counts every 10 residues in human Slo3. ( B ) Alignments of human Slo1 and Slo3 from various species are shown for the cytosolic gating ring domain beginning with the conserved sequence at the beginning of the first RCK domain. Blue highlights residue differences between human Slo1 and human Slo3. Yellow highlights differences of Slo3 of various species to human Slo3. Alignments were generated by Clustal 1.2.0 and minor adjustments were made based on structural considerations ( Leonetti et al., 2012 ). Above the residues, the correspondence of particular amino acid segments to structurally defined α-helical and β-strand segments is shown based on Leonetti et al. (2012) . In red, residues or segments identified in Slo1 or Slo3 isoforms which are implicated in ligand-sensing or species-specific functional differences are highlighted. Although extensive information is available regarding loci important in ligand-sensing in Slo1, such information for Slo3 remains lacking. Numbers identify the following: 1, the sequence of residues termed the Ca 2+ bowl ( Schreiber and Salkoff, 1997 ), for which there is good correspondence of mutations affecting Ca2+-dependent function ( Bao et al., 2004 ) and coordination of density in a crystal structure ( Yuan et al., 2012 ); 2, the D367 residue implicated in the role of the RCK1 domain in Ca 2+ -dependent activation ( Xia et al., 2002 ) which is clearly distinct from Ca 2+ -bowl dependent activation ( Zeng et al., 2005 ); 3, the M513 residue, which also affects Ca 2+− dependent activation involving the RCK1 domain ( Bao et al., 2002 ), but probably is not involved in ligand coordination; 4, residues E374 and E399 which have been implicated in low affinity effects of divalent cations, specifically Mg 2+ ( Shi et al., 2002 ; Xia et al., 2002 ; Yang et al., 2006 ); 5, residue E535 which may also be involved in Ca 2+ coordination in RCK1 ( Zhang et al., 2010 ); 6, residues H365 and H394, which have been implicated in proton-dependent activation of Slo1 and also influence Ca 2+ -dependent activation when protonated ( Hou et al., 2008 ); 7, H417 and segment 368–475, which influence pH-sensing in mouse Slo3 (Zhang et al., 2006); 8, segment 495–515 in bovine Slo3 which accounts for part of the different in functional properties between mouse Slo3 and bovine Slo3 ( Santi et al., 2009 ). Illustrated sequences and accession numbers include: HsSlo1 ( Homo sapiens ), NP_001154824, Gene ID 3778; HsSlo3 ( Homo sapiens ), NP_001027006, Gene ID 157855; MmSlo3 ( Mus musculus ), NP_032458, Gene ID 16532; RnSlo3 ( Rattus norvegicus ), XP_006253398, Gene ID 680912; TcSlo3 (Tupaia chinensis, Chinese tree shrew), XP_006171561, Gene ID 102493286; CcSlo3 ( Condylura cristata , star-nosed mole), XP_004682520, Gene ID 101620543; CfSlo3 (Canis lupus familiaris), XP_539971, Gene ID 482856; BtSlo3 (Bos taurus), NP_001156721, Gene ID 524144; OaSlo3 ( Ovis aries , sheep), XP_004021821, Gene ID 10110209. DOI: http://dx.doi.org/10.7554/eLife.01438.019

    Techniques Used: Sequencing, Generated, Functional Assay, Activation Assay

    2) Product Images from "Inositol 1,4,5-Trisphosphate Directs Ca2+ Flow between Mitochondria and the Endoplasmic/Sarcoplasmic Reticulum: A Role in Regulating Cardiac Autonomic Ca2+ Spiking"

    Article Title: Inositol 1,4,5-Trisphosphate Directs Ca2+ Flow between Mitochondria and the Endoplasmic/Sarcoplasmic Reticulum: A Role in Regulating Cardiac Autonomic Ca2+ Spiking

    Journal: Molecular Biology of the Cell

    doi:

    Intracellular Localization and Properties of the IP3 -sensitive Intracellular Ca2+ Pool
    Figure Legend Snippet: Intracellular Localization and Properties of the IP3 -sensitive Intracellular Ca2+ Pool

    Techniques Used:

    3) Product Images from "Small mouse cholangiocytes proliferate in response to H1 histamine receptor stimulation by activation of the IP3/CaMK I/CREB pathway"

    Article Title: Small mouse cholangiocytes proliferate in response to H1 histamine receptor stimulation by activation of the IP3/CaMK I/CREB pathway

    Journal:

    doi: 10.1152/ajpcell.00369.2007

    Effect of HTMT Dimaleate on Intracellular IP3 , Ca2+ , and cAMP Levels
    Figure Legend Snippet: Effect of HTMT Dimaleate on Intracellular IP3 , Ca2+ , and cAMP Levels

    Techniques Used:

    4) Product Images from "INVOLVEMENT OF PHOSPHOLIPASE C SIGNALING IN MELANOMA CELL-INDUCED ENDOTHELIAL JUNCTION DISASSEMBLY"

    Article Title: INVOLVEMENT OF PHOSPHOLIPASE C SIGNALING IN MELANOMA CELL-INDUCED ENDOTHELIAL JUNCTION DISASSEMBLY

    Journal:

    doi:

    Pertussis toxin inhibits endothelial [Ca2+]i responses. HUVEC were preincubated with 0.5 μg/ml PT in culture medium for 2 hr. Melanoma cell contact induced an increase 121 ± 22 % (n = 11) in peak [Ca2+]i magnitude in untreated endothelial
    Figure Legend Snippet: Pertussis toxin inhibits endothelial [Ca2+]i responses. HUVEC were preincubated with 0.5 μg/ml PT in culture medium for 2 hr. Melanoma cell contact induced an increase 121 ± 22 % (n = 11) in peak [Ca2+]i magnitude in untreated endothelial

    Techniques Used:

    Endothelial cells release [Ca2+]i in response to contact with melanoma cells. Melanoma cells elicited a threefold increase of [Ca2+]i over baseline in HUVEC. In contrast, inert beads did not trigger significant [Ca2+]i response. Arrows indicate the time
    Figure Legend Snippet: Endothelial cells release [Ca2+]i in response to contact with melanoma cells. Melanoma cells elicited a threefold increase of [Ca2+]i over baseline in HUVEC. In contrast, inert beads did not trigger significant [Ca2+]i response. Arrows indicate the time

    Techniques Used:

    5) Product Images from "Allopregnanolone-induced rise in intracellular calcium in embryonic hippocampal neurons parallels their proliferative potential"

    Article Title: Allopregnanolone-induced rise in intracellular calcium in embryonic hippocampal neurons parallels their proliferative potential

    Journal: BMC Neuroscience

    doi: 10.1186/1471-2202-9-S2-S11

    APα induces a rapid and transient [Ca 2+ ] i rise in rat E18 hippocampal neurons in primary culture. Cells grown on glass cover slips were loaded with Fura-2 AM as described in the Methods. The [Ca 2+ ] i was determined on single cells in HBSS medium. (A) Images represent calcium fluro-2 fluorescence in rat hippocampal neurons under vehicle control (top) and 500 nM APα (bottom). A fluorescent gradient of fluorescence ratio of 340 nm/380 nm is presented at the right side of the images. (B) A linear regression and the multiple correlation coefficient R-square value indicate the perfect correlation between fluorescence ratio of 340 nm/380 nm and [Ca 2+ ]. (C) APα induced rapid and transient responses of [Ca 2+ ] i recorded in a time course and expressed as the fluorescence ratio between 340 nm/380 nm, which is a representative of three different experiments. (D) Bar graph represents the summary of [Ca2+] i in response to control, vehicle control, and 500 nM APα. Data are mean ± SEM (* p
    Figure Legend Snippet: APα induces a rapid and transient [Ca 2+ ] i rise in rat E18 hippocampal neurons in primary culture. Cells grown on glass cover slips were loaded with Fura-2 AM as described in the Methods. The [Ca 2+ ] i was determined on single cells in HBSS medium. (A) Images represent calcium fluro-2 fluorescence in rat hippocampal neurons under vehicle control (top) and 500 nM APα (bottom). A fluorescent gradient of fluorescence ratio of 340 nm/380 nm is presented at the right side of the images. (B) A linear regression and the multiple correlation coefficient R-square value indicate the perfect correlation between fluorescence ratio of 340 nm/380 nm and [Ca 2+ ]. (C) APα induced rapid and transient responses of [Ca 2+ ] i recorded in a time course and expressed as the fluorescence ratio between 340 nm/380 nm, which is a representative of three different experiments. (D) Bar graph represents the summary of [Ca2+] i in response to control, vehicle control, and 500 nM APα. Data are mean ± SEM (* p

    Techniques Used: Fluorescence

    6) Product Images from "INVOLVEMENT OF PHOSPHOLIPASE C SIGNALING IN MELANOMA CELL-INDUCED ENDOTHELIAL JUNCTION DISASSEMBLY"

    Article Title: INVOLVEMENT OF PHOSPHOLIPASE C SIGNALING IN MELANOMA CELL-INDUCED ENDOTHELIAL JUNCTION DISASSEMBLY

    Journal:

    doi:

    Pertussis toxin inhibits endothelial [Ca2+]i responses. HUVEC were preincubated with 0.5 μg/ml PT in culture medium for 2 hr. Melanoma cell contact induced an increase 121 ± 22 % (n = 11) in peak [Ca2+]i magnitude in untreated endothelial
    Figure Legend Snippet: Pertussis toxin inhibits endothelial [Ca2+]i responses. HUVEC were preincubated with 0.5 μg/ml PT in culture medium for 2 hr. Melanoma cell contact induced an increase 121 ± 22 % (n = 11) in peak [Ca2+]i magnitude in untreated endothelial

    Techniques Used:

    Endothelial cells release [Ca2+]i in response to contact with melanoma cells. Melanoma cells elicited a threefold increase of [Ca2+]i over baseline in HUVEC. In contrast, inert beads did not trigger significant [Ca2+]i response. Arrows indicate the time
    Figure Legend Snippet: Endothelial cells release [Ca2+]i in response to contact with melanoma cells. Melanoma cells elicited a threefold increase of [Ca2+]i over baseline in HUVEC. In contrast, inert beads did not trigger significant [Ca2+]i response. Arrows indicate the time

    Techniques Used:

    7) Product Images from "The PDZ Scaffold NHERF-2 Interacts with mGluR5 and Regulates Receptor Activity"

    Article Title: The PDZ Scaffold NHERF-2 Interacts with mGluR5 and Regulates Receptor Activity

    Journal:

    doi: 10.1074/jbc.M602262200

    NHERF-2 Selectively Prolongs Agonist-induced Ca2+ Signaling by mGluR5
    Figure Legend Snippet: NHERF-2 Selectively Prolongs Agonist-induced Ca2+ Signaling by mGluR5

    Techniques Used:

    8) Product Images from "Gene expression pattern of functional neuronal cells derived from human bone marrow mesenchymal stromal cells"

    Article Title: Gene expression pattern of functional neuronal cells derived from human bone marrow mesenchymal stromal cells

    Journal: BMC Genomics

    doi: 10.1186/1471-2164-9-166

    Intracellular Calcium influx . Representative variation of intracellular Calcium influx in differentiated MSC treated with AMPA (40 μM, ---×---), glutamate (100 μM, ) or CNQX (10 μM, ). Results are expressed as fluorescence intensity at 4 second-intervals using the Fluo-4 NW tracer. Agonists (AMPA or Glutamate) and antagonist CNQX were compared with spontaneous level of calcium influx in presence of Ca2+ alone ( ).
    Figure Legend Snippet: Intracellular Calcium influx . Representative variation of intracellular Calcium influx in differentiated MSC treated with AMPA (40 μM, ---×---), glutamate (100 μM, ) or CNQX (10 μM, ). Results are expressed as fluorescence intensity at 4 second-intervals using the Fluo-4 NW tracer. Agonists (AMPA or Glutamate) and antagonist CNQX were compared with spontaneous level of calcium influx in presence of Ca2+ alone ( ).

    Techniques Used: Fluorescence

    9) Product Images from "NCX1 represents an ionic Na+ sensing mechanism in macrophages"

    Article Title: NCX1 represents an ionic Na+ sensing mechanism in macrophages

    Journal: PLoS Biology

    doi: 10.1371/journal.pbio.3000722

    HS exposure causes Ca 2+ loss. (A) Relative [Ca 2+ ] i levels of RAW264.7 MΦs. Traces of Fura-2–loaded RAW264.7 MΦs stimulated ± HS at t = 10 s (mean ± SD; n = 6). Where indicated, Tg was added (means ± SD; n = 2). (B) As in (A), but RAW264.7 MΦs were stimulated with LPS ± HS at t = 10 s (mean ± SD; n = 5). Where indicated, Tg was added (means ± SD; n = 2). (C) As in (A), but relative [Ca 2+ ] i levels were assessed in Fluo-3/Fura-Red–loaded MΦs (means ± SD; n = 6). (D) As in (B), but relative [Ca 2+ ] i levels were assessed in Fluo-3/Fura-Red–loaded MΦs (means ± SD; n = 8). For numerical raw data, please see S1 Data . [Ca2+] i , intracellular Ca 2+ in situ; HS, high salt; LPS, lipopolysaccharide; MΦ, monocyte/macrophage-like cell; Tg, thapsigargin.
    Figure Legend Snippet: HS exposure causes Ca 2+ loss. (A) Relative [Ca 2+ ] i levels of RAW264.7 MΦs. Traces of Fura-2–loaded RAW264.7 MΦs stimulated ± HS at t = 10 s (mean ± SD; n = 6). Where indicated, Tg was added (means ± SD; n = 2). (B) As in (A), but RAW264.7 MΦs were stimulated with LPS ± HS at t = 10 s (mean ± SD; n = 5). Where indicated, Tg was added (means ± SD; n = 2). (C) As in (A), but relative [Ca 2+ ] i levels were assessed in Fluo-3/Fura-Red–loaded MΦs (means ± SD; n = 6). (D) As in (B), but relative [Ca 2+ ] i levels were assessed in Fluo-3/Fura-Red–loaded MΦs (means ± SD; n = 8). For numerical raw data, please see S1 Data . [Ca2+] i , intracellular Ca 2+ in situ; HS, high salt; LPS, lipopolysaccharide; MΦ, monocyte/macrophage-like cell; Tg, thapsigargin.

    Techniques Used: In Situ

    10) Product Images from "Calsequestrin 2 (CASQ2) mutations increase expression of calreticulin and ryanodine receptors, causing catecholaminergic polymorphic ventricular tachycardia"

    Article Title: Calsequestrin 2 (CASQ2) mutations increase expression of calreticulin and ryanodine receptors, causing catecholaminergic polymorphic ventricular tachycardia

    Journal:

    doi: 10.1172/JCI31080

    SR Ca2+ and contractility in isolated CASQ2-deficient myocytes.
    Figure Legend Snippet: SR Ca2+ and contractility in isolated CASQ2-deficient myocytes.

    Techniques Used: Isolation

    11) Product Images from "Ca2+ responses of pulmonary arterial myocytes to acute hypoxia require release from ryanodine and inositol trisphosphate receptors in sarcoplasmic reticulum"

    Article Title: Ca2+ responses of pulmonary arterial myocytes to acute hypoxia require release from ryanodine and inositol trisphosphate receptors in sarcoplasmic reticulum

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    doi: 10.1152/ajplung.00348.2011

    Compartmentation of SR Ca2+ stores in rat distal PASMC.
    Figure Legend Snippet: Compartmentation of SR Ca2+ stores in rat distal PASMC.

    Techniques Used:

    12) Product Images from "Inositol 1,4,5-Trisphosphate Directs Ca2+ Flow between Mitochondria and the Endoplasmic/Sarcoplasmic Reticulum: A Role in Regulating Cardiac Autonomic Ca2+ Spiking"

    Article Title: Inositol 1,4,5-Trisphosphate Directs Ca2+ Flow between Mitochondria and the Endoplasmic/Sarcoplasmic Reticulum: A Role in Regulating Cardiac Autonomic Ca2+ Spiking

    Journal: Molecular Biology of the Cell

    doi:

    Intracellular Localization and Properties of the IP3 -sensitive Intracellular Ca2+ Pool
    Figure Legend Snippet: Intracellular Localization and Properties of the IP3 -sensitive Intracellular Ca2+ Pool

    Techniques Used:

    13) Product Images from "Determinants in CaV1 Channels That Regulate the Ca2+ Sensitivity of Bound Calmodulin *"

    Article Title: Determinants in CaV1 Channels That Regulate the Ca2+ Sensitivity of Bound Calmodulin *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M109.013326

    Partially Ca2+ -saturated CaM Binds with Both Lobes to CaV 1.2 IQ
    Figure Legend Snippet: Partially Ca2+ -saturated CaM Binds with Both Lobes to CaV 1.2 IQ

    Techniques Used: Chick Chorioallantoic Membrane Assay

    Amino Acids in the IQ Motif That Regulate the Ca2+ Affinity of Bound CaM
    Figure Legend Snippet: Amino Acids in the IQ Motif That Regulate the Ca2+ Affinity of Bound CaM

    Techniques Used: Chick Chorioallantoic Membrane Assay

    Partially Ca2+ -saturated CaM Binds with Both Lobes to CaV 1.2 IQ
    Figure Legend Snippet: Partially Ca2+ -saturated CaM Binds with Both Lobes to CaV 1.2 IQ

    Techniques Used: Chick Chorioallantoic Membrane Assay

    14) Product Images from "Rat Strain Differences in Pulmonary Artery Smooth Muscle Ca2+ Entry Following Chronic Hypoxia"

    Article Title: Rat Strain Differences in Pulmonary Artery Smooth Muscle Ca2+ Entry Following Chronic Hypoxia

    Journal:

    doi: 10.1080/10739680903114268

    In Situ Calibrations of VSM [Ca2+ ]i
    Figure Legend Snippet: In Situ Calibrations of VSM [Ca2+ ]i

    Techniques Used: In Situ

    Basal VSM [Ca2+ ]i
    Figure Legend Snippet: Basal VSM [Ca2+ ]i

    Techniques Used:

    15) Product Images from "Ion Channels and Transporters in Lung Function and Disease: Hydrogen peroxide-induced calcium influx in lung microvascular endothelial cells involves TRPV4"

    Article Title: Ion Channels and Transporters in Lung Function and Disease: Hydrogen peroxide-induced calcium influx in lung microvascular endothelial cells involves TRPV4

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    doi: 10.1152/ajplung.00275.2015

    Identifying the source of Ca2+ in H2 O2 -induced responses.
    Figure Legend Snippet: Identifying the source of Ca2+ in H2 O2 -induced responses.

    Techniques Used:

    16) Product Images from "Allele and species dependent contractile defects by restrictive and hypertrophic cardiomyopathy-linked troponin I mutants"

    Article Title: Allele and species dependent contractile defects by restrictive and hypertrophic cardiomyopathy-linked troponin I mutants

    Journal:

    doi: 10.1016/j.yjmcc.2008.02.274

    3.3 RCM linked mutations in cTnI’s Helix-4/C-terminal domain slow relaxation and Ca2+ transient decay rate
    Figure Legend Snippet: 3.3 RCM linked mutations in cTnI’s Helix-4/C-terminal domain slow relaxation and Ca2+ transient decay rate

    Techniques Used:

    17) Product Images from "Pu-Erh Tea Relaxes the Thoracic Aorta of Rats by Reducing Intracellular Calcium"

    Article Title: Pu-Erh Tea Relaxes the Thoracic Aorta of Rats by Reducing Intracellular Calcium

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2019.01430

    The study schematic representing the mechanism of pu-erh tea-induced vasodilation in the rat thoracic aorta. Pu-erh aqueous extract vasodilated arteries in an endothelium-independent manner, which might partly by inhibiting extracellular Ca2+ influx. Additionally, TBs and CAF should be the main active components. TBs, theabrownins; CAF, caffeine.
    Figure Legend Snippet: The study schematic representing the mechanism of pu-erh tea-induced vasodilation in the rat thoracic aorta. Pu-erh aqueous extract vasodilated arteries in an endothelium-independent manner, which might partly by inhibiting extracellular Ca2+ influx. Additionally, TBs and CAF should be the main active components. TBs, theabrownins; CAF, caffeine.

    Techniques Used:

    18) Product Images from "Ca2+ responses of pulmonary arterial myocytes to acute hypoxia require release from ryanodine and inositol trisphosphate receptors in sarcoplasmic reticulum"

    Article Title: Ca2+ responses of pulmonary arterial myocytes to acute hypoxia require release from ryanodine and inositol trisphosphate receptors in sarcoplasmic reticulum

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    doi: 10.1152/ajplung.00348.2011

    Compartmentation of SR Ca2+ stores in rat distal PASMC.
    Figure Legend Snippet: Compartmentation of SR Ca2+ stores in rat distal PASMC.

    Techniques Used:

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    Article Snippet: .. For transduction, the cells were washed twice with PBS containing Mg2+ and Ca2+ (Invitrogen). .. Then, 0.3 ml of the medium containing the virus particles was added.

    Transfection:

    Article Title: MicroRNA-203 functions as a tumor suppressor in basal cell carcinoma
    Article Snippet: .. Transfections Human adult skin epidermal keratinocytes (obtained from Cascade Biologics, Portland, OR, USA) were cultured in EpiLife serum-free keratinocyte growth medium, including Human Keratinocyte Growth Supplement at a final Ca2+ concentration of 0.06 mM (Cascade Biologics). .. Third passage keratinocytes at 70% confluence were transfected with 10 nM Pre-miR 203 miRNA Precursor (Ambion Applied Biosystems, Austin, TX, USA) or 10 nM Pre-miR miRNA Precursor Negative Control #1 (Ambion) using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA), following the manufacturer's instruction.

    Cell Culture:

    Article Title: MicroRNA-203 functions as a tumor suppressor in basal cell carcinoma
    Article Snippet: .. Transfections Human adult skin epidermal keratinocytes (obtained from Cascade Biologics, Portland, OR, USA) were cultured in EpiLife serum-free keratinocyte growth medium, including Human Keratinocyte Growth Supplement at a final Ca2+ concentration of 0.06 mM (Cascade Biologics). .. Third passage keratinocytes at 70% confluence were transfected with 10 nM Pre-miR 203 miRNA Precursor (Ambion Applied Biosystems, Austin, TX, USA) or 10 nM Pre-miR miRNA Precursor Negative Control #1 (Ambion) using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA), following the manufacturer's instruction.

    Article Title: Inhibition of Keratinocyte Differentiation by the Synergistic Effect of IL-17A, IL-22, IL-1α, TNFα and Oncostatin M
    Article Snippet: .. NHEK were cultured to 80% of confluence allowing the expression of a large panel of keratinocyte differentiation markers, and then starved for 24 h in Keratinocyte SFM containing 0.03 mM Ca2+ (Invitrogen Life Technologies, Cergy Pontoise, France) before stimulation. .. Confluent differentiated cells were stimulated with or without recombinant IL-17A, OSM, TNFα, IL-22 and IL-1α alone at maximum effective concentrations (reported previously around 10 ng/ml , ) or in combination (R & D systems Europe, Lille, France) during 2 h to 72 h for mRNA quantification.

    Article Title: Exosomal Release of L-Plastin by Breast Cancer Cells Facilitates Metastatic Bone Osteolysis
    Article Snippet: .. On days 2-3, cells were supplemented with fresh media with or without RANKL (50 ng/ml) or recombinant L-plastin (rP2, 2.5-25 μg/ml) +/− a [Ca2+ ]i chelator BAPTA-acetoxymethyl ester (6-50 μM BAPTA, Invitrogen, B6769) for 10 minutes as previously described , washed, treated with recombinant L-plastin (rP2, 2.5-25 μg/ml), cultured for 2 days, fixed, and stained for TRAP. ..

    Concentration Assay:

    Article Title: MicroRNA-203 functions as a tumor suppressor in basal cell carcinoma
    Article Snippet: .. Transfections Human adult skin epidermal keratinocytes (obtained from Cascade Biologics, Portland, OR, USA) were cultured in EpiLife serum-free keratinocyte growth medium, including Human Keratinocyte Growth Supplement at a final Ca2+ concentration of 0.06 mM (Cascade Biologics). .. Third passage keratinocytes at 70% confluence were transfected with 10 nM Pre-miR 203 miRNA Precursor (Ambion Applied Biosystems, Austin, TX, USA) or 10 nM Pre-miR miRNA Precursor Negative Control #1 (Ambion) using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA), following the manufacturer's instruction.

    other:

    Article Title: Sequential forward and reverse transport of the Na+ Ca2+ exchanger generates Ca2+ oscillations within mitochondria
    Article Snippet: siRNA against rat mitofusin 2 was from Invitrogen (stealth RNAiTM 5193986). siRNA against human mitofusin 2 was from Origene (Cat No: SR306670). siRNA against OPA1 was from Origene (Cat No: SR505373). siRNA against MCU was from Origene (Cat No: SR508660). siRNA against the rat mitochondrial Na+ –Ca2+ exchanger was from Invitrogen (AACGGCCACUCAACUGUCU) and human mitochondrial Na+ –Ca2+ exchanger was from Origene (Cat No: SR312772).

    Article Title: Sequential forward and reverse transport of the Na+ Ca2+ exchanger generates Ca2+ oscillations within mitochondria
    Article Snippet: siRNA knockdown siRNA against rat mitofusin 2 was from Invitrogen (stealth RNAiTM 5193986). siRNA against human mitofusin 2 was from Origene (Cat No: SR306670). siRNA against OPA1 was from Origene (Cat No: SR505373). siRNA against MCU was from Origene (Cat No: SR508660). siRNA against the rat mitochondrial Na+ –Ca2+ exchanger was from Invitrogen (AACGGCCACUCAACUGUCU) and human mitochondrial Na+ –Ca2+ exchanger was from Origene (Cat No: SR312772).

    Article Title: SIRT1 Promotes Differentiation of Normal Human Keratinocytes
    Article Snippet: NHEK cells obtained from Cascade Biologics (Portland, OR), and maintained under standard conditions in Epilife Medium with Ca2+ (Cascade Biologics) and supplemented with human keratinocyte growth supplement (Cascade Biologics).

    Expressing:

    Article Title: Inhibition of Keratinocyte Differentiation by the Synergistic Effect of IL-17A, IL-22, IL-1α, TNFα and Oncostatin M
    Article Snippet: .. NHEK were cultured to 80% of confluence allowing the expression of a large panel of keratinocyte differentiation markers, and then starved for 24 h in Keratinocyte SFM containing 0.03 mM Ca2+ (Invitrogen Life Technologies, Cergy Pontoise, France) before stimulation. .. Confluent differentiated cells were stimulated with or without recombinant IL-17A, OSM, TNFα, IL-22 and IL-1α alone at maximum effective concentrations (reported previously around 10 ng/ml , ) or in combination (R & D systems Europe, Lille, France) during 2 h to 72 h for mRNA quantification.

    Staining:

    Article Title: Exosomal Release of L-Plastin by Breast Cancer Cells Facilitates Metastatic Bone Osteolysis
    Article Snippet: .. On days 2-3, cells were supplemented with fresh media with or without RANKL (50 ng/ml) or recombinant L-plastin (rP2, 2.5-25 μg/ml) +/− a [Ca2+ ]i chelator BAPTA-acetoxymethyl ester (6-50 μM BAPTA, Invitrogen, B6769) for 10 minutes as previously described , washed, treated with recombinant L-plastin (rP2, 2.5-25 μg/ml), cultured for 2 days, fixed, and stained for TRAP. ..

    Recombinant:

    Article Title: Exosomal Release of L-Plastin by Breast Cancer Cells Facilitates Metastatic Bone Osteolysis
    Article Snippet: .. On days 2-3, cells were supplemented with fresh media with or without RANKL (50 ng/ml) or recombinant L-plastin (rP2, 2.5-25 μg/ml) +/− a [Ca2+ ]i chelator BAPTA-acetoxymethyl ester (6-50 μM BAPTA, Invitrogen, B6769) for 10 minutes as previously described , washed, treated with recombinant L-plastin (rP2, 2.5-25 μg/ml), cultured for 2 days, fixed, and stained for TRAP. ..

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    Thermo Fisher l type ca2 channel
    CASQ2D307H myocytes show increased stability of <t>Ca2+</t> cycling when challenged with a β-adrenergic agonist.
    L Type Ca2 Channel, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher mitochondrial ca2
    Computer model of mouse ventricular cell. A, Schematic diagrams of the 3‐dimensional structure of the cell model (left) and the Ca 2+ release unit ( CRU )–mitochondrial Ca 2+ cycling model (right). B, The modified L‐type Ca 2+ current model. Left: The Hodgkin‐Huxley ( HH ) scheme. Right: The equivalent Markov scheme of the HH scheme. To simulate a much lower channel open probability (≈5%–10%) observed in experiments, we added a new state (the final open state), with the opening rate from the d 2 f 2 f <t>Ca2</t> state being r 1 and the closing rate being r 2 . CYTO indicates cytosolic space; DS, dyadic space; jm‐NaCa, mitochondrial Na + ‐Ca 2+ exchanger Ca2+ release; JSR, junctional sarcoplasmic reticulum; j uni , mitochondrial Ca 2+ uniporter uptake; j up , sarco/endoplasmic reticulum Ca 2+ ‐ATPase (SERCA) uptake; LCC, L‐type Ca 2+ channel; MITO, mitochondrial space; NSR, network sarcoplasmic reticulum; RyR, ryanodine receptor; SUB, submembrane space.
    Mitochondrial Ca2, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher ca2 ca1 pyramidal neurons
    RGS14 expression blocks long-term spine plasticity in <t>CA2</t> and <t>CA1</t> neurons lacking RGS14, and high extracellular Ca 2+ restores structural plasticity to RGS14-expressing neurons. A , Averaged time course of spine volume change in the presence or absence of RGS14 during the induction of spine structural plasticity by repetitive two-photon glutamate uncaging in the absence of extracellular Mg 2+ in CA2 pyramidal neurons from RGS14 KO mice. The number of samples (spines/neurons/animals) for stimulated spines are 11/4/3 for GFP (black) and 11/4/3 for RGS14-GFP (red). B , Quantification of the average volume change for stimulated spines either in the presence or absence of RGS14 during the transient (1–3-min) and sustained (21–25-min) phases of sLTP induction for CA2 neurons. Unpaired t test, **, p = 0.01. C , Averaged time course of spine volume change during the induction of spine structural plasticity in the presence or absence of RGS14 by repetitive two-photon glutamate uncaging in the absence of extracellular Mg 2+ in CA1 pyramidal neurons from RGS14 KO mice. The number of samples (spines/neurons/animals) for stimulated spines are 11/4/4 for GFP (black) and 23/9/6 for RGS14-GFP 4mM external [Ca 2+ ] (red), and 9/4/4 for RGS14-GFP external 8mM [Ca 2+ ] (blue). D , Quantification of the average volume change for stimulated spines in the presence or absence of RGS14 during the transient and sustained phases of sLTP induction for CA1 neurons. Fisher’s LSD test, *, p
    Ca2 Ca1 Pyramidal Neurons, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    CASQ2D307H myocytes show increased stability of Ca2+ cycling when challenged with a β-adrenergic agonist.

    Journal: American Journal of Physiology - Heart and Circulatory Physiology

    Article Title: Functional consequences of stably expressing a mutant calsequestrin (CASQ2D307H) in the CASQ2 null background

    doi: 10.1152/ajpheart.00578.2011

    Figure Lengend Snippet: CASQ2D307H myocytes show increased stability of Ca2+ cycling when challenged with a β-adrenergic agonist.

    Article Snippet: Expression of SR proteins SERCA2a (Zymed), phospholamban (PLB, Zymed), TRD1 (Santa Cruz), CASQ2 (Thermo Scientific), L-type Ca2+ channel (DHPR2α, Thermo Scientific), RyR2 (Thermo Scientific), phosphorylated PLB (serine 16 and threonine 17; Badrilla), and GAPDH (Cell Signaling) were determined by quantitative Western blotting techniques ( ) using whole heart homogenates from WT, CASQ2D307H , and CASQ2 null ( n = 6).

    Techniques:

    Computer model of mouse ventricular cell. A, Schematic diagrams of the 3‐dimensional structure of the cell model (left) and the Ca 2+ release unit ( CRU )–mitochondrial Ca 2+ cycling model (right). B, The modified L‐type Ca 2+ current model. Left: The Hodgkin‐Huxley ( HH ) scheme. Right: The equivalent Markov scheme of the HH scheme. To simulate a much lower channel open probability (≈5%–10%) observed in experiments, we added a new state (the final open state), with the opening rate from the d 2 f 2 f Ca2 state being r 1 and the closing rate being r 2 . CYTO indicates cytosolic space; DS, dyadic space; jm‐NaCa, mitochondrial Na + ‐Ca 2+ exchanger Ca2+ release; JSR, junctional sarcoplasmic reticulum; j uni , mitochondrial Ca 2+ uniporter uptake; j up , sarco/endoplasmic reticulum Ca 2+ ‐ATPase (SERCA) uptake; LCC, L‐type Ca 2+ channel; MITO, mitochondrial space; NSR, network sarcoplasmic reticulum; RyR, ryanodine receptor; SUB, submembrane space.

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Mitochondrial Ca2+ Influx Contributes to Arrhythmic Risk in Nonischemic Cardiomyopathy

    doi: 10.1161/JAHA.117.007805

    Figure Lengend Snippet: Computer model of mouse ventricular cell. A, Schematic diagrams of the 3‐dimensional structure of the cell model (left) and the Ca 2+ release unit ( CRU )–mitochondrial Ca 2+ cycling model (right). B, The modified L‐type Ca 2+ current model. Left: The Hodgkin‐Huxley ( HH ) scheme. Right: The equivalent Markov scheme of the HH scheme. To simulate a much lower channel open probability (≈5%–10%) observed in experiments, we added a new state (the final open state), with the opening rate from the d 2 f 2 f Ca2 state being r 1 and the closing rate being r 2 . CYTO indicates cytosolic space; DS, dyadic space; jm‐NaCa, mitochondrial Na + ‐Ca 2+ exchanger Ca2+ release; JSR, junctional sarcoplasmic reticulum; j uni , mitochondrial Ca 2+ uniporter uptake; j up , sarco/endoplasmic reticulum Ca 2+ ‐ATPase (SERCA) uptake; LCC, L‐type Ca 2+ channel; MITO, mitochondrial space; NSR, network sarcoplasmic reticulum; RyR, ryanodine receptor; SUB, submembrane space.

    Article Snippet: Mitochondrial Ca2+ transients were monitored by loading cells with Rhod‐2 AM (Thermo Fisher Scientific; 1 μmol/L; 1 hour, 37°C) combined with the ruptured current‐clamp technique with 20 μmol/L EGTA added in the pipette solution.

    Techniques: Modification

    RGS14 expression blocks long-term spine plasticity in CA2 and CA1 neurons lacking RGS14, and high extracellular Ca 2+ restores structural plasticity to RGS14-expressing neurons. A , Averaged time course of spine volume change in the presence or absence of RGS14 during the induction of spine structural plasticity by repetitive two-photon glutamate uncaging in the absence of extracellular Mg 2+ in CA2 pyramidal neurons from RGS14 KO mice. The number of samples (spines/neurons/animals) for stimulated spines are 11/4/3 for GFP (black) and 11/4/3 for RGS14-GFP (red). B , Quantification of the average volume change for stimulated spines either in the presence or absence of RGS14 during the transient (1–3-min) and sustained (21–25-min) phases of sLTP induction for CA2 neurons. Unpaired t test, **, p = 0.01. C , Averaged time course of spine volume change during the induction of spine structural plasticity in the presence or absence of RGS14 by repetitive two-photon glutamate uncaging in the absence of extracellular Mg 2+ in CA1 pyramidal neurons from RGS14 KO mice. The number of samples (spines/neurons/animals) for stimulated spines are 11/4/4 for GFP (black) and 23/9/6 for RGS14-GFP 4mM external [Ca 2+ ] (red), and 9/4/4 for RGS14-GFP external 8mM [Ca 2+ ] (blue). D , Quantification of the average volume change for stimulated spines in the presence or absence of RGS14 during the transient and sustained phases of sLTP induction for CA1 neurons. Fisher’s LSD test, *, p

    Journal: eNeuro

    Article Title: RGS14 Restricts Plasticity in Hippocampal CA2 by Limiting Postsynaptic Calcium Signaling

    doi: 10.1523/ENEURO.0353-17.2018

    Figure Lengend Snippet: RGS14 expression blocks long-term spine plasticity in CA2 and CA1 neurons lacking RGS14, and high extracellular Ca 2+ restores structural plasticity to RGS14-expressing neurons. A , Averaged time course of spine volume change in the presence or absence of RGS14 during the induction of spine structural plasticity by repetitive two-photon glutamate uncaging in the absence of extracellular Mg 2+ in CA2 pyramidal neurons from RGS14 KO mice. The number of samples (spines/neurons/animals) for stimulated spines are 11/4/3 for GFP (black) and 11/4/3 for RGS14-GFP (red). B , Quantification of the average volume change for stimulated spines either in the presence or absence of RGS14 during the transient (1–3-min) and sustained (21–25-min) phases of sLTP induction for CA2 neurons. Unpaired t test, **, p = 0.01. C , Averaged time course of spine volume change during the induction of spine structural plasticity in the presence or absence of RGS14 by repetitive two-photon glutamate uncaging in the absence of extracellular Mg 2+ in CA1 pyramidal neurons from RGS14 KO mice. The number of samples (spines/neurons/animals) for stimulated spines are 11/4/4 for GFP (black) and 23/9/6 for RGS14-GFP 4mM external [Ca 2+ ] (red), and 9/4/4 for RGS14-GFP external 8mM [Ca 2+ ] (blue). D , Quantification of the average volume change for stimulated spines in the presence or absence of RGS14 during the transient and sustained phases of sLTP induction for CA1 neurons. Fisher’s LSD test, *, p

    Article Snippet: Calcium imaging For Ca2+ imaging, we performed whole-cell patch recordings of CA2/CA1 pyramidal neurons in cultured hippocampus slices, with the patch pipette containing the Ca2+ indicator Fluo-4FF (500 µm ; Thermo Fisher) and Alexa Fluor 594 (500 µm ) diluted in potassium gluconate internal solution (containing in mm : 130 K gluconate, 10 Na phosphocreatine, 4 MgCl2 , 4 Na2 ATP, 0.3 MgGTP, l -ascorbic acid 3, HEPES 10, pH 7.4, 300 mOsm).

    Techniques: Expressing, Mouse Assay

    RGS14 KO mice exhibit plasticity of CA2 synapses into adulthood. A , Amigo2-EGFP fluorescence (green) labels CA2 pyramidal neurons as shown by overlap with another CA2 molecular marker PCP4 (red). Scale bar = 100 μm. B , Amigo2-EGFP fluorescence (green) does not colocalize with immunoreactivity for the CA1 pyramidal neuron marker WFS1 (magenta). Scale bar = 100 μm. C , Summary graph of field potential recordings from acute hippocampal slices prepared from adult RGS14 WT and KO mice (both Amigo2-EGFP+) validate that RGS14 KO mice possess a capacity for LTP in CA2 in adulthood (red), which is absent in WT mice (purple). RGS14 WT and KO mice do not differ in CA1 plasticity (green, gray). LTP was induced by high-frequency stimulation (HFS; 3 × 100 Hz) at time 0 (arrow). Data are represented as mean normalized fEPSP slope ± SEM (dotted reference line at 1.0). Sample sizes (in slices/animals) are WT CA2 n = 14/5; KO CA2 n = 16/4; WT CA1 n = 17/4; KO CA1 n = 12/5. Insets (top) are representative traces of field potentials recorded from areas CA2 and CA1 from slices of RGS14 WT and KO mice before (light line) and after (heavy line) LTP induction. D , Quantification of the mean normalized fEPSP slope 40-60 min following LTP induction ( C ) with error bars representing SEM. The difference in the degree of LTP induced in RGS14 WT and KO CA2 synapses was significant, whereas no difference was detected between RGS14 WT and KO synapses recorded in CA1. Sidak’s, **, p ≤ 0.01, ns = not significant.

    Journal: eNeuro

    Article Title: RGS14 Restricts Plasticity in Hippocampal CA2 by Limiting Postsynaptic Calcium Signaling

    doi: 10.1523/ENEURO.0353-17.2018

    Figure Lengend Snippet: RGS14 KO mice exhibit plasticity of CA2 synapses into adulthood. A , Amigo2-EGFP fluorescence (green) labels CA2 pyramidal neurons as shown by overlap with another CA2 molecular marker PCP4 (red). Scale bar = 100 μm. B , Amigo2-EGFP fluorescence (green) does not colocalize with immunoreactivity for the CA1 pyramidal neuron marker WFS1 (magenta). Scale bar = 100 μm. C , Summary graph of field potential recordings from acute hippocampal slices prepared from adult RGS14 WT and KO mice (both Amigo2-EGFP+) validate that RGS14 KO mice possess a capacity for LTP in CA2 in adulthood (red), which is absent in WT mice (purple). RGS14 WT and KO mice do not differ in CA1 plasticity (green, gray). LTP was induced by high-frequency stimulation (HFS; 3 × 100 Hz) at time 0 (arrow). Data are represented as mean normalized fEPSP slope ± SEM (dotted reference line at 1.0). Sample sizes (in slices/animals) are WT CA2 n = 14/5; KO CA2 n = 16/4; WT CA1 n = 17/4; KO CA1 n = 12/5. Insets (top) are representative traces of field potentials recorded from areas CA2 and CA1 from slices of RGS14 WT and KO mice before (light line) and after (heavy line) LTP induction. D , Quantification of the mean normalized fEPSP slope 40-60 min following LTP induction ( C ) with error bars representing SEM. The difference in the degree of LTP induced in RGS14 WT and KO CA2 synapses was significant, whereas no difference was detected between RGS14 WT and KO synapses recorded in CA1. Sidak’s, **, p ≤ 0.01, ns = not significant.

    Article Snippet: Calcium imaging For Ca2+ imaging, we performed whole-cell patch recordings of CA2/CA1 pyramidal neurons in cultured hippocampus slices, with the patch pipette containing the Ca2+ indicator Fluo-4FF (500 µm ; Thermo Fisher) and Alexa Fluor 594 (500 µm ) diluted in potassium gluconate internal solution (containing in mm : 130 K gluconate, 10 Na phosphocreatine, 4 MgCl2 , 4 Na2 ATP, 0.3 MgGTP, l -ascorbic acid 3, HEPES 10, pH 7.4, 300 mOsm).

    Techniques: Mouse Assay, Fluorescence, Marker

    RGS14 suppresses CA2 spine structural plasticity. A , Representative post hoc immunostaining to delineate hippocampal region CA2 region after imaging biolistically transfected neurons. Left: Organotypic hippocampus slice culture stained for the DG- and CA2-enriched gene PCP4 (red). Scale bar = 100 µm. Right: Magnified view of area CA2 in PCP4 immunostained (red) hippocampus on left with a biolistically labeled CA2 pyramidal neuron expressing mEGFP (green). Scale bar = 50 µm. B , Averaged time course of spine volume change during the induction of spine structural plasticity (sLTP) by repetitive two-photon glutamate uncaging (top bar; 30 pulses at 0.5 Hz) in the absence of extracellular Mg 2+ . The number of samples (spines/neurons/animals) for stimulated spines are 19/6/5 for WT CA2, 15/8/6 for KO CA2, 23/7/5 for WT CA1, and 16/6/5 for KO CA1. Sample size applies to B and C . Error bars denote SEM. C , Quantification of the average volume change for stimulated spines during the transient (1–3-min) and sustained (21–25-min) phases of sLTP induction. Sidak’s, *, p

    Journal: eNeuro

    Article Title: RGS14 Restricts Plasticity in Hippocampal CA2 by Limiting Postsynaptic Calcium Signaling

    doi: 10.1523/ENEURO.0353-17.2018

    Figure Lengend Snippet: RGS14 suppresses CA2 spine structural plasticity. A , Representative post hoc immunostaining to delineate hippocampal region CA2 region after imaging biolistically transfected neurons. Left: Organotypic hippocampus slice culture stained for the DG- and CA2-enriched gene PCP4 (red). Scale bar = 100 µm. Right: Magnified view of area CA2 in PCP4 immunostained (red) hippocampus on left with a biolistically labeled CA2 pyramidal neuron expressing mEGFP (green). Scale bar = 50 µm. B , Averaged time course of spine volume change during the induction of spine structural plasticity (sLTP) by repetitive two-photon glutamate uncaging (top bar; 30 pulses at 0.5 Hz) in the absence of extracellular Mg 2+ . The number of samples (spines/neurons/animals) for stimulated spines are 19/6/5 for WT CA2, 15/8/6 for KO CA2, 23/7/5 for WT CA1, and 16/6/5 for KO CA1. Sample size applies to B and C . Error bars denote SEM. C , Quantification of the average volume change for stimulated spines during the transient (1–3-min) and sustained (21–25-min) phases of sLTP induction. Sidak’s, *, p

    Article Snippet: Calcium imaging For Ca2+ imaging, we performed whole-cell patch recordings of CA2/CA1 pyramidal neurons in cultured hippocampus slices, with the patch pipette containing the Ca2+ indicator Fluo-4FF (500 µm ; Thermo Fisher) and Alexa Fluor 594 (500 µm ) diluted in potassium gluconate internal solution (containing in mm : 130 K gluconate, 10 Na phosphocreatine, 4 MgCl2 , 4 Na2 ATP, 0.3 MgGTP, l -ascorbic acid 3, HEPES 10, pH 7.4, 300 mOsm).

    Techniques: Immunostaining, Imaging, Transfection, Staining, Labeling, Expressing

    RGS14 restricts Ca 2+ levels in CA2 spines. A , Averaged time courses of spine Ca 2+ transients measured with Fluo-4FF during two-photon glutamate uncaging to induce sLTP (30 pulses, 0.5 Hz). Data are shown as the average change in spine Ca 2+ concentration (Δ[Ca 2+ ]) from CA2 pyramidal neurons. B , Averaged time courses of spine Ca 2+ transients measured with Fluo-4FF and Alexa Fluor 594 during two-photon glutamate uncaging to induce sLTP (30 pulses, 0.5 Hz). Data are shown as the average change in spine Ca 2+ (Δ[Ca 2+ ]) from CA1 pyramidal neurons. C , Uncaging-triggered averages for the change in spine Ca 2+ concentration during sLTP induction. Error bars denote SEM. D , RGS14 limits CA2 spine Ca 2+ transients. Bar graphs displaying the average peak Δ[Ca 2+ ] ± SEM (****, p

    Journal: eNeuro

    Article Title: RGS14 Restricts Plasticity in Hippocampal CA2 by Limiting Postsynaptic Calcium Signaling

    doi: 10.1523/ENEURO.0353-17.2018

    Figure Lengend Snippet: RGS14 restricts Ca 2+ levels in CA2 spines. A , Averaged time courses of spine Ca 2+ transients measured with Fluo-4FF during two-photon glutamate uncaging to induce sLTP (30 pulses, 0.5 Hz). Data are shown as the average change in spine Ca 2+ concentration (Δ[Ca 2+ ]) from CA2 pyramidal neurons. B , Averaged time courses of spine Ca 2+ transients measured with Fluo-4FF and Alexa Fluor 594 during two-photon glutamate uncaging to induce sLTP (30 pulses, 0.5 Hz). Data are shown as the average change in spine Ca 2+ (Δ[Ca 2+ ]) from CA1 pyramidal neurons. C , Uncaging-triggered averages for the change in spine Ca 2+ concentration during sLTP induction. Error bars denote SEM. D , RGS14 limits CA2 spine Ca 2+ transients. Bar graphs displaying the average peak Δ[Ca 2+ ] ± SEM (****, p

    Article Snippet: Calcium imaging For Ca2+ imaging, we performed whole-cell patch recordings of CA2/CA1 pyramidal neurons in cultured hippocampus slices, with the patch pipette containing the Ca2+ indicator Fluo-4FF (500 µm ; Thermo Fisher) and Alexa Fluor 594 (500 µm ) diluted in potassium gluconate internal solution (containing in mm : 130 K gluconate, 10 Na phosphocreatine, 4 MgCl2 , 4 Na2 ATP, 0.3 MgGTP, l -ascorbic acid 3, HEPES 10, pH 7.4, 300 mOsm).

    Techniques: Concentration Assay