Multiple quantum filtered 23Na NMR in the Langendorff perfused mouse heart: Ratio of triple/double quantum filtered signals correlates with [Na]i
Identifieur interne : 000373 ( Pmc/Curation ); précédent : 000372; suivant : 000374Multiple quantum filtered 23Na NMR in the Langendorff perfused mouse heart: Ratio of triple/double quantum filtered signals correlates with [Na]i
Auteurs : Thomas R. Eykyn [Royaume-Uni] ; Dunja Aksentijevi [Royaume-Uni] ; Karen L. Aughton [Royaume-Uni] ; Richard Southworth [Royaume-Uni] ; William Fuller [Royaume-Uni] ; Michael J. Shattock [Royaume-Uni]Source :
- Journal of Molecular and Cellular Cardiology [ 0022-2828 ] ; 2015.
Abstract
We investigate the potential of multiple quantum filtered (MQF) 23Na NMR to probe intracellular [Na]i in the Langendorff perfused mouse heart. In the presence of Tm(DOTP) shift reagent the triple quantum filtered (TQF) signal originated largely from the intracellular sodium pool with a 32 ± 6% contribution of the total TQF signal arising from extracellular sodium, whilst the rank 2 double-quantum filtered signal (DQF), acquired with a 54.7° flip-angle pulse, originated exclusively from the extracellular sodium pool. Given the different cellular origins of the 23Na MQF signals we propose that the TQF/DQF ratio can be used as a semi-quantitative measure of [Na]i in the mouse heart. We demonstrate a good correlation of this ratio with [Na]i measured with shift reagent at baseline and under conditions of elevated [Na]i. We compare the measurements of [Na]i using both shift reagent and TQF/DQF ratio in a cohort of wild type mouse hearts and in a transgenic PLM3SA mouse expressing a non-phosphorylatable form of phospholemman, showing a modest but measurable elevation of baseline [Na]i. MQF filtered 23Na NMR is a potentially useful tool for studying normal and pathophysiological changes in [Na]i, particularly in transgenic mouse models with altered Na regulation.
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DOI: 10.1016/j.yjmcc.2015.07.009
PubMed: 26196304
PubMed Central: 4564289
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Eykyn</name><affiliation wicri:level="1"><nlm:aff id="af0005">Department of Imaging Chemistry and Biology, Division of Imaging Sciences and Biomedical Engineering, King's College London, King's Health Partners, St. Thomas' Hospital, London SE1 7EH, United Kingdom</nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Department of Imaging Chemistry and Biology, Division of Imaging Sciences and Biomedical Engineering, King's College London, King's Health Partners, St. Thomas' Hospital, London SE1 7EH</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="af0010">The British Heart Foundation Centre of Research Excellence, The Rayne Institute, King's College London, St. Thomas' Hospital, London SE1 7EH, United Kingdom</nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>The British Heart Foundation Centre of Research Excellence, The Rayne Institute, King's College London, St. Thomas' Hospital, London SE1 7EH</wicri:regionArea></affiliation></author><author><name sortKey="Aksentijevi, Dunja" sort="Aksentijevi, Dunja" uniqKey="Aksentijevi D" first="Dunja" last="Aksentijevi">Dunja Aksentijevi</name><affiliation wicri:level="1"><nlm:aff id="af0010">The British Heart Foundation Centre of Research Excellence, The Rayne Institute, King's College London, St. Thomas' Hospital, London SE1 7EH, United Kingdom</nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>The British Heart Foundation Centre of Research Excellence, The Rayne Institute, King's College London, St. Thomas' Hospital, London SE1 7EH</wicri:regionArea></affiliation></author><author><name sortKey="Aughton, Karen L" sort="Aughton, Karen L" uniqKey="Aughton K" first="Karen L." last="Aughton">Karen L. 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Eykyn</name><affiliation wicri:level="1"><nlm:aff id="af0005">Department of Imaging Chemistry and Biology, Division of Imaging Sciences and Biomedical Engineering, King's College London, King's Health Partners, St. Thomas' Hospital, London SE1 7EH, United Kingdom</nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Department of Imaging Chemistry and Biology, Division of Imaging Sciences and Biomedical Engineering, King's College London, King's Health Partners, St. Thomas' Hospital, London SE1 7EH</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="af0010">The British Heart Foundation Centre of Research Excellence, The Rayne Institute, King's College London, St. Thomas' Hospital, London SE1 7EH, United Kingdom</nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>The British Heart Foundation Centre of Research Excellence, The Rayne Institute, King's College London, St. Thomas' Hospital, London SE1 7EH</wicri:regionArea></affiliation></author><author><name sortKey="Aksentijevi, Dunja" sort="Aksentijevi, Dunja" uniqKey="Aksentijevi D" first="Dunja" last="Aksentijevi">Dunja Aksentijevi</name><affiliation wicri:level="1"><nlm:aff id="af0010">The British Heart Foundation Centre of Research Excellence, The Rayne Institute, King's College London, St. Thomas' Hospital, London SE1 7EH, United Kingdom</nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>The British Heart Foundation Centre of Research Excellence, The Rayne Institute, King's College London, St. Thomas' Hospital, London SE1 7EH</wicri:regionArea></affiliation></author><author><name sortKey="Aughton, Karen L" sort="Aughton, Karen L" uniqKey="Aughton K" first="Karen L." last="Aughton">Karen L. Aughton</name><affiliation wicri:level="1"><nlm:aff id="af0010">The British Heart Foundation Centre of Research Excellence, The Rayne Institute, King's College London, St. Thomas' Hospital, London SE1 7EH, United Kingdom</nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>The British Heart Foundation Centre of Research Excellence, The Rayne Institute, King's College London, St. Thomas' Hospital, London SE1 7EH</wicri:regionArea></affiliation></author><author><name sortKey="Southworth, Richard" sort="Southworth, Richard" uniqKey="Southworth R" first="Richard" last="Southworth">Richard Southworth</name><affiliation wicri:level="1"><nlm:aff id="af0005">Department of Imaging Chemistry and Biology, Division of Imaging Sciences and Biomedical Engineering, King's College London, King's Health Partners, St. Thomas' Hospital, London SE1 7EH, United Kingdom</nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Department of Imaging Chemistry and Biology, Division of Imaging Sciences and Biomedical Engineering, King's College London, King's Health Partners, St. Thomas' Hospital, London SE1 7EH</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="af0010">The British Heart Foundation Centre of Research Excellence, The Rayne Institute, King's College London, St. Thomas' Hospital, London SE1 7EH, United Kingdom</nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>The British Heart Foundation Centre of Research Excellence, The Rayne Institute, King's College London, St. Thomas' Hospital, London SE1 7EH</wicri:regionArea></affiliation></author><author><name sortKey="Fuller, William" sort="Fuller, William" uniqKey="Fuller W" first="William" last="Fuller">William Fuller</name><affiliation wicri:level="1"><nlm:aff id="af0015">Division of Cardiovascular and Diabetes Medicine, University of Dundee, Dundee, United Kingdom</nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Division of Cardiovascular and Diabetes Medicine, University of Dundee, Dundee</wicri:regionArea></affiliation></author><author><name sortKey="Shattock, Michael J" sort="Shattock, Michael J" uniqKey="Shattock M" first="Michael J." last="Shattock">Michael J. Shattock</name><affiliation wicri:level="1"><nlm:aff id="af0010">The British Heart Foundation Centre of Research Excellence, The Rayne Institute, King's College London, St. Thomas' Hospital, London SE1 7EH, United Kingdom</nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>The British Heart Foundation Centre of Research Excellence, The Rayne Institute, King's College London, St. Thomas' Hospital, London SE1 7EH</wicri:regionArea></affiliation></author></analytic><series><title level="j">Journal of Molecular and Cellular Cardiology</title><idno type="ISSN">0022-2828</idno><idno type="eISSN">1095-8584</idno><imprint><date when="2015">2015</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>We investigate the potential of multiple quantum filtered (MQF) <sup>23</sup>Na NMR to probe intracellular [Na]<sub>i</sub> in the Langendorff perfused mouse heart. In the presence of Tm(DOTP) shift reagent the triple quantum filtered (TQF) signal originated largely from the intracellular sodium pool with a 32 ± 6% contribution of the total TQF signal arising from extracellular sodium, whilst the rank 2 double-quantum filtered signal (DQF), acquired with a 54.7° flip-angle pulse, originated exclusively from the extracellular sodium pool. Given the different cellular origins of the <sup>23</sup>Na MQF signals we propose that the TQF/DQF ratio can be used as a semi-quantitative measure of [Na]<sub>i</sub> in the mouse heart. We demonstrate a good correlation of this ratio with [Na]<sub>i</sub> measured with shift reagent at baseline and under conditions of elevated [Na]<sub>i</sub>. We compare the measurements of [Na]<sub>i</sub> using both shift reagent and TQF/DQF ratio in a cohort of wild type mouse hearts and in a transgenic PLM<sup>3SA</sup> mouse expressing a non-phosphorylatable form of phospholemman, showing a modest but measurable elevation of baseline [Na]<sub>i</sub>. MQF filtered <sup>23</sup>Na NMR is a potentially useful tool for studying normal and pathophysiological changes in [Na]<sub>i</sub>, particularly in transgenic mouse models with altered Na regulation.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Murphy, E" uniqKey="Murphy E">E. Murphy</name></author><author><name sortKey="Eisner, D A" uniqKey="Eisner D">D.A. Eisner</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Despa, S" uniqKey="Despa S">S. Despa</name></author><author><name sortKey="Bers, D M" uniqKey="Bers D">D.M. 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<front><journal-meta><journal-id journal-id-type="nlm-ta">J Mol Cell Cardiol</journal-id><journal-id journal-id-type="iso-abbrev">J. Mol. Cell. Cardiol</journal-id><journal-title-group><journal-title>Journal of Molecular and Cellular Cardiology</journal-title></journal-title-group><issn pub-type="ppub">0022-2828</issn><issn pub-type="epub">1095-8584</issn><publisher><publisher-name>Academic Press</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">26196304</article-id><article-id pub-id-type="pmc">4564289</article-id><article-id pub-id-type="publisher-id">S0022-2828(15)30012-2</article-id><article-id pub-id-type="doi">10.1016/j.yjmcc.2015.07.009</article-id><article-categories><subj-group subj-group-type="heading"><subject>Original Article</subject></subj-group></article-categories><title-group><article-title>Multiple quantum filtered <sup>23</sup>Na NMR in the Langendorff perfused mouse heart: Ratio of triple/double quantum filtered signals correlates with [Na]<sub>i</sub></article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Eykyn</surname><given-names>Thomas R.</given-names></name><email>thomas.eykyn@kcl.ac.uk</email><xref rid="af0005" ref-type="aff">a</xref><xref rid="af0010" ref-type="aff">b</xref><xref rid="cr0005" ref-type="corresp">⁎</xref><xref rid="fn0005" ref-type="fn">1</xref></contrib><contrib contrib-type="author"><name><surname>Aksentijević</surname><given-names>Dunja</given-names></name><xref rid="af0010" ref-type="aff">b</xref><xref rid="fn0005" ref-type="fn">1</xref></contrib><contrib contrib-type="author"><name><surname>Aughton</surname><given-names>Karen L.</given-names></name><xref rid="af0010" ref-type="aff">b</xref></contrib><contrib contrib-type="author"><name><surname>Southworth</surname><given-names>Richard</given-names></name><xref rid="af0005" ref-type="aff">a</xref><xref rid="af0010" ref-type="aff">b</xref></contrib><contrib contrib-type="author"><name><surname>Fuller</surname><given-names>William</given-names></name><xref rid="af0015" ref-type="aff">c</xref></contrib><contrib contrib-type="author"><name><surname>Shattock</surname><given-names>Michael J.</given-names></name><xref rid="af0010" ref-type="aff">b</xref></contrib></contrib-group><aff id="af0005"><label>a</label>Department of Imaging Chemistry and Biology, Division of Imaging Sciences and Biomedical Engineering, King's College London, King's Health Partners, St. Thomas' Hospital, London SE1 7EH, United Kingdom</aff><aff id="af0010"><label>b</label>The British Heart Foundation Centre of Research Excellence, The Rayne Institute, King's College London, St. Thomas' Hospital, London SE1 7EH, United Kingdom</aff><aff id="af0015"><label>c</label>Division of Cardiovascular and Diabetes Medicine, University of Dundee, Dundee, United Kingdom</aff><author-notes><corresp id="cr0005"><label>⁎</label>Corresponding author at: Department of Imaging Chemistry and Biology, Division of Imaging Sciences and Biomedical Engineering, King's College London, King's Health Partners, St. Thomas' Hospital, London SE1 7EH, United Kingdom. <email>thomas.eykyn@kcl.ac.uk</email></corresp><fn id="fn0005"><label>1</label><p>Authors contributed equally.</p></fn></author-notes><pub-date pub-type="pmc-release"><day>1</day><month>9</month><year>2015</year></pub-date><pmc-comment> PMC Release delay is 0 months and 0 days and was based on .</pmc-comment>
<pub-date pub-type="ppub"><month>9</month><year>2015</year></pub-date><volume>86</volume><fpage>95</fpage><lpage>101</lpage><history><date date-type="received"><day>21</day><month>3</month><year>2015</year></date><date date-type="rev-recd"><day>8</day><month>7</month><year>2015</year></date><date date-type="accepted"><day>10</day><month>7</month><year>2015</year></date></history><permissions><copyright-statement>© 2015 The Authors. Published by Elsevier Ltd.</copyright-statement><copyright-year>2015</copyright-year><copyright-holder></copyright-holder><license license-type="CC BY" xlink:href="http://creativecommons.org/licenses/by/4.0/"><license-p>This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).</license-p></license></permissions><abstract><p>We investigate the potential of multiple quantum filtered (MQF) <sup>23</sup>Na NMR to probe intracellular [Na]<sub>i</sub> in the Langendorff perfused mouse heart. In the presence of Tm(DOTP) shift reagent the triple quantum filtered (TQF) signal originated largely from the intracellular sodium pool with a 32 ± 6% contribution of the total TQF signal arising from extracellular sodium, whilst the rank 2 double-quantum filtered signal (DQF), acquired with a 54.7° flip-angle pulse, originated exclusively from the extracellular sodium pool. Given the different cellular origins of the <sup>23</sup>Na MQF signals we propose that the TQF/DQF ratio can be used as a semi-quantitative measure of [Na]<sub>i</sub> in the mouse heart. We demonstrate a good correlation of this ratio with [Na]<sub>i</sub> measured with shift reagent at baseline and under conditions of elevated [Na]<sub>i</sub>. We compare the measurements of [Na]<sub>i</sub> using both shift reagent and TQF/DQF ratio in a cohort of wild type mouse hearts and in a transgenic PLM<sup>3SA</sup> mouse expressing a non-phosphorylatable form of phospholemman, showing a modest but measurable elevation of baseline [Na]<sub>i</sub>. MQF filtered <sup>23</sup>Na NMR is a potentially useful tool for studying normal and pathophysiological changes in [Na]<sub>i</sub>, particularly in transgenic mouse models with altered Na regulation.</p></abstract><abstract abstract-type="author-highlights"><title>Highlights</title><p><list list-type="simple"><list-item id="u0005"><label>•</label><p>Intracellular Na concentration [Na]<sub>i</sub> is a key modulator of cardiac cell function.</p></list-item><list-item id="u0010"><label>•</label><p>We developed an NMR-compatible Langendorff mouse heart perfusion system.</p></list-item><list-item id="u0015"><label>•</label><p>The ratio of triple/double quantum filtered <sup>23</sup>Na NMR signals correlates with [Na]<sub>i</sub>.</p></list-item><list-item id="u0020"><label>•</label><p>Intracellular [Na]<sub>i</sub> can be quantified under physiological perfusion conditions.</p></list-item><list-item id="u0025"><label>•</label><p>The PLM<sup>3SA</sup> transgenic mouse model has a measurable elevation of [Na]<sub>i</sub> at baseline.</p></list-item></list></p></abstract><kwd-group><title>Keywords</title><kwd>Multiple quantum filtered <sup>23</sup>Na</kwd><kwd>TQF</kwd><kwd>DQF</kwd><kwd>Langendorff perfused mouse heart</kwd><kwd>Shift reagent</kwd></kwd-group></article-meta></front><floats-group><fig id="f0005"><label>Fig. 1</label><caption><p>NMR compatible Langendorff constant pressure perfusion system (a, b) and a schematic of the pneumatically driven three-way valve to allow switching between reservoirs with minimum dead-volume (c).</p></caption><graphic xlink:href="gr1"></graphic></fig><fig id="f0010"><label>Fig. 2</label><caption><p>(a) Example <sup>31</sup>P spectrum acquired during baseline stability period showing normal cardiac energetics. (b) Time-series of <sup>23</sup>Na NMR spectra acquired in a perfused mouse heart during the infusion of Tm(DOTP) shift reagent. An initial shift of the intravascular extracellular signal [Na]<sub>e</sub> is observed, followed by the signal from the buffer surrounding the heart [Na]<sub>o</sub> was achieved within a 5–10 min timeframe.</p></caption><graphic xlink:href="gr2"></graphic></fig><fig id="f0015"><label>Fig. 3</label><caption><p>(a) <sup>23</sup>Na single quantum spectrum post-infusion of shift reagent where the intracellular peak from [Na]<sub>i</sub> is clearly discerned. (b) TQF spectrum acquired from the same heart as in (a) in the presence of shift reagent showing predominantly intracellular [Na]<sub>i</sub> with a smaller contribution from extracellular [Na]<sub>e</sub>. (c) <sup>23</sup>Na DQF spectrum acquired with a flip angle β = 54.7° showing predominantly extracellular [Na]<sub>e</sub>. (d) <sup>23</sup>Na DQF spectrum acquired with a flip angle β = 90° showing both intracellular and extracellular Na.</p></caption><graphic xlink:href="gr3"></graphic></fig><fig id="f0020"><label>Fig. 4</label><caption><p>(a) Peak areas shown for a time-series (± S.E. n = 6) of <sup>23</sup>Na TQF and DQF spectra acquired in an interleaved fashion in the perfused mouse heart during stability and during subsequent infusion of 50 μM ouabain, shaded area. Representative TQF and DQF spectra are shown; (b) during stability, (c) after 20 min infusion of ouabain, (d) after 20 min K<sup>+</sup>-free buffer and (e) after 20 min K<sup>+</sup>-free/Ca<sup>2 +</sup>-free buffer.</p></caption><graphic xlink:href="gr4"></graphic></fig><fig id="f0025"><label>Fig. 5</label><caption><p>Plot of the TQF/DQF ratio R<sup>TQF/DQF</sup> vs the measured concentration [Na]<sub>i</sub> in the perfused mouse heart at baseline (filled square, n = 6) and following a 20 min intervention to elevate [Na]<sub>i</sub>; 50 μM ouabain (filled triangle, n = 6); 0 mM K<sup>+</sup> (open diamond, n = 3), 0 mM K<sup>+</sup>/0 mM Ca<sup>2 +</sup> (filled circle, n = 3) and 0 mM K<sup>+</sup>/0 mM Ca<sup>2 +</sup>/0 mM Mg<sup>2 +</sup> (open circle, n = 3). Mean data are shown for each group ± S.E.</p></caption><graphic xlink:href="gr5"></graphic></fig><fig id="f0030"><label>Fig. 6</label><caption><p>Plot of measured [Na]<sub>i</sub> in the WT mouse and PLM<sup>3SA</sup> transgenic mouse using (a) the shift reagent technique and (b) the derived values of [Na]<sub>i</sub> using the TQF/DQF ratio. Significant differences between the two groups are shown **p < 0.05.</p></caption><graphic xlink:href="gr6"></graphic></fig><table-wrap id="t0005" position="float"><label>Table 1</label><caption><p>Cardiac function acquired at 5 min intervals during the 20 min stability period.</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="left">Functional parameters<hr></hr></th><th colspan="5" align="left">Time (min)<hr></hr></th></tr><tr><th align="left">(n = 7)</th><th align="left">0</th><th align="left">5</th><th align="left">10</th><th align="left">15</th><th align="left">20</th></tr></thead><tbody><tr><td align="left">Systolic pressure (mm Hg)</td><td align="char">109 ± 12</td><td align="char">113 ± 11</td><td align="char">103 ± 17</td><td align="char">103 ± 19</td><td align="char">106 ± 19</td></tr><tr><td align="left">EDP (mm Hg)</td><td align="char">11 ± 1</td><td align="char">9 ± 2</td><td align="char">11 ± 2</td><td align="char">10 ± 2</td><td align="char">12 ± 2</td></tr><tr><td align="left">LVDP (mm Hg)</td><td align="char">98 ± 12</td><td align="char">104 ± 11</td><td align="char">92 ± 16</td><td align="char">93 ± 18</td><td align="char">93 ± 19</td></tr><tr><td align="left">Heart rate (bpm)</td><td align="char">382 ± 32</td><td align="char">526 ± 54</td><td align="char">453 ± 67</td><td align="char">449 ± 79</td><td align="char">464 ± 96</td></tr><tr><td align="left">Coronary flow (ml/min)</td><td align="char">2.2 ± 0.1</td><td align="char">2.2 ± 0.2</td><td align="char">2.4 ± 0.2</td><td align="char">2.2 ± 0.2</td><td align="char">2.6 ± 0.2</td></tr></tbody></table><table-wrap-foot><fn><p>EDP — end diastolic pressure, LVDP — left ventricular developed pressure.</p></fn></table-wrap-foot></table-wrap></floats-group></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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