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FHF2KO and Wild-Type Mouse Cardiomyocyte Strands (Park et al 2020)
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mso 9]><xml> <o:shapedefaults v:ext="edit" spidmax="1026"/> </xml><![endif]--><!--[if gte mso 9]><xml> <o:shapelayout v:ext="edit"> <o:idmap v:ext="edit" data="1"/> </o:shapelayout></xml><![endif]--> </head> <body lang=EN-US style='tab-interval:36.0pt'> <div class=WordSection1> <p class=MsoNormal align=center style='text-align:center'><b style='mso-bidi-font-weight: normal'><span style='font-size:11.0pt'>Description and Use of <i style='mso-bidi-font-style:normal'>Fhf2<sup>WT</sup></i> and <i style='mso-bidi-font-style:normal'>Fhf2<sup>KO</sup></i> Cardiomyocyte Strand Models<o:p></o:p></span></b></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><b style='mso-bidi-font-weight:normal'><span style='font-size:11.0pt'><o:p> </o:p></span></b></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><b style='mso-bidi-font-weight:normal'><span style='font-size:11.0pt'>DESCRIPTION: </span></b><span style='font-size:11.0pt'>The <i style='mso-bidi-font-style: normal'>Fhf2<sup>WT</sup></i> and <i style='mso-bidi-font-style:normal'>Fhf2<sup>KO</sup></i> ventricular cardiomyocyte models described previously (Park et al, <i style='mso-bidi-font-style:normal'>Nature Comm.</i> 7:12966, 2016) were scripted onto the NEURON software platform (Hines and <span class=SpellE>Carnevale</span>, <i style='mso-bidi-font-style:normal'>Neuroscientist</i> 7:123, 2001) with several modifications and then linked into strands with gap <span class=SpellE>junctional</span> <span class=SpellE>conductances</span>.<o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><b style='mso-bidi-font-weight:normal'><i style='mso-bidi-font-style:normal'><span style='font-size:11.0pt;font-family:Times'><o:p> </o:p></span></i></b></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><b style='mso-bidi-font-weight:normal'><i style='mso-bidi-font-style:normal'><span style='font-size:11.0pt;font-family:Times'>Cardiomyocyte Dimensions and Gap Junction Connectivity into Strands:</span></i></b><span style='font-size:11.0pt; font-family:Times'><span style="mso-spacerun:yes"> </span>The cardiomyocyte model cells (<span class=SpellE>myocyte.hoc</span>) are cylinders with length (<span class=SpellE><span class=GramE>cell.L</span></span>) of 100 microns and diameter (<span class=SpellE>cell.diam</span>) of 22.34 microns. With standard membrane unit capacitance (cell.cm) of 1 </span><span style='font-size:11.0pt; font-family:Symbol'>m</span><span style='font-size:11.0pt;font-family:Times'>F/cm<sup>2</sup>, each cell’s membrane capacitance is 70 <span class=SpellE>pF.</span><span style="mso-spacerun:yes"> </span>The number of cells in the strand can be selected in the Graphical User Interface (GUI); all published data analysis was on strands comprising 111 cells (1.11 cm length).<o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times'><o:p> </o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times'>Gap junctions are modeled as reciprocal and equivalently weighted <span class=SpellE>conductances</span> between adjacent cells n and n+1.<span style="mso-spacerun:yes"> </span>The conductance in <q>source</q> cell n is <span class=SpellE>gap_<span class=GramE>sources.o</span></span><span class=GramE>[</span>n].g, while the matching conductance in <q>sink</q> cell n+1 is <span class=SpellE>gap_dests.o</span>[n+1].g, with the currents driven by the voltage differential between source cell n and sink cell n+1.<span style="mso-spacerun:yes"> </span>These <span class=SpellE>conductances</span> are absolute set values (in <span class=SpellE>pS</span>) multiplied by Q10 = </span><span style='font-size:11.0pt'>1.43<sup>{(<span style='position:relative;top:-3.0pt;mso-text-raise:3.0pt'>o</span>C – 37)/10}</sup></span><span style='font-size:11.0pt;font-family:Times'>, unlike <span class=SpellE>transmembrane</span> ion <span class=SpellE>conductances</span> (below), which are expressed as conductance densities (S/cm<sup>2</sup>).<span style="mso-spacerun:yes"> </span>The normal physiological setting for <span class=SpellE>junctional</span> conductance is 772.8 <span class=SpellE>nS</span> at 37<sup>o</sup>C.<span style="mso-spacerun:yes"> </span>The <span class=SpellE>junctional</span> <span class=SpellE>conductances</span> can be manipulated equally between all cell pairs in the GUI, or specific cell pair <span class=SpellE>conductances</span> can be varied by text commands.<o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times'><o:p> </o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><b style='mso-bidi-font-weight:normal'><i style='mso-bidi-font-style:normal'><span style='font-size:11.0pt;font-family:Times'>Ion <span class=SpellE>Conductances</span> in <span class=SpellE>Cardiomyocytes</span>:</span></i></b><b style='mso-bidi-font-weight: normal'><span style='font-size:11.0pt;font-family:Times'><span style="mso-spacerun:yes"> </span></span></b><span style='font-size:11.0pt; font-family:Times'>All ion conductance densities are set equivalently in all cells of the strand.<span style="mso-spacerun:yes"> </span>Each ion conductance density can be manipulated equally throughout the strand in the GUI, or specific cell ion conductance densities can be varied by text commands.<o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times'><o:p> </o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><u><span style='font-size:11.0pt;font-family:Times'>Voltage-gated sodium <span class=SpellE>conductances</span>:</span></u><span style='font-size:11.0pt; font-family:Times'><span style="mso-spacerun:yes"> </span>The <span class=SpellE>myocytes</span> include two 16-state Markov model voltage-dependent sodium <span class=SpellE>conductances</span> termed <span class=SpellE>NAV_withF</span> and <span class=SpellE>NAV_noF</span> (Scheme 1).<span style="mso-spacerun:yes"> </span><o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><u><span style='font-size:11.0pt;font-family:Times'>Scheme 1<o:p></o:p></span></u></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span lang=IT style='font-size:11.0pt;font-family:Times;mso-ansi-language:IT; mso-fareast-language:IT;mso-no-proof:yes'><!--[if gte vml 1]><v:shapetype id="_x0000_t75" coordsize="21600,21600" o:spt="75" o:preferrelative="t" path="m@4@5l@4@11@9@11@9@5xe" filled="f" stroked="f"> <v:stroke joinstyle="miter"/> <v:formulas> <v:f eqn="if lineDrawn pixelLineWidth 0"/> <v:f eqn="sum @0 1 0"/> <v:f eqn="sum 0 0 @1"/> <v:f eqn="prod @2 1 2"/> <v:f eqn="prod @3 21600 pixelWidth"/> <v:f eqn="prod @3 21600 pixelHeight"/> <v:f eqn="sum @0 0 1"/> <v:f eqn="prod @6 1 2"/> <v:f eqn="prod @7 21600 pixelWidth"/> <v:f eqn="sum @8 21600 0"/> <v:f eqn="prod @7 21600 pixelHeight"/> <v:f eqn="sum @10 21600 0"/> </v:formulas> <v:path o:extrusionok="f" gradientshapeok="t" o:connecttype="rect"/> <o:lock v:ext="edit" aspectratio="t"/> </v:shapetype><v:shape id="Picture_x0020_1" o:spid="_x0000_i1026" type="#_x0000_t75" style='width:6in;height:151pt;visibility:visible;mso-wrap-style:square'> <v:imagedata src="README_file/image001.png" o:title=""/> </v:shape><![endif]--><![if !vml]><img width=434 height=153 src="README_file/image002.png" v:shapes="Picture_x0020_1"><![endif]></span><span style='font-size:11.0pt;font-family:Times'><o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><i style='mso-bidi-font-style:normal'><span style='font-size:11.0pt;font-family: Times'><o:p> </o:p></span></i></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><i style='mso-bidi-font-style:normal'><span style='font-size:11.0pt;font-family: Times'>Fhf2<sup>KO</sup></span></i><span style='font-size:11.0pt;font-family: Times'> <span class=SpellE>cardiomyocytes</span> only have a functional <span class=SpellE>NAV_noF</span> conductance (i.e. </span><!--[if gte msEquation 12]><m:oMath><m:acc><m:accPr><m:chr m:val="̅"/><span style='font-size:11.0pt;mso-ansi-font-size:11.0pt; mso-bidi-font-size:11.0pt;font-family:"Cambria Math";mso-ascii-font-family: "Cambria Math";mso-hansi-font-family:"Cambria Math";font-style:italic; mso-bidi-font-style:normal'><m:ctrlPr></m:ctrlPr></span></m:accPr><m:e><span style='font-size:11.0pt;font-family:Times'><m:r><span><m:rPr><m:nor/></m:rPr>gNav</m:r></span></span></m:e></m:acc></m:oMath><![endif]--><![if !msEquation]><span style='font-size:12.0pt;font-family:"Times New Roman";mso-fareast-font-family: "Times New Roman";position:relative;top:2.0pt;mso-text-raise:-2.0pt;mso-ansi-language: EN-US;mso-fareast-language:EN-US;mso-bidi-language:AR-SA'><img width=26 height=15 id="_x0000_i1025" src="README_file/image003.png"></span><![endif]><span style='font-size:11.0pt;font-family:Times'><span style="mso-spacerun:yes"> </span>for <span class=SpellE>NAV_withF</span> = 0), while <i style='mso-bidi-font-style:normal'>Fhf2<sup>WT</sup></i> <span class=SpellE>cardiomyocytes</span> contain a mixture of <span class=SpellE>NAV_withF</span> and <span class=SpellE>NAV_noF</span>.<span style="mso-spacerun:yes"> </span>Employing this mixture does not imply knowledge that wild-type ventricular <span class=SpellE>cardiomyocytes</span> necessarily bear a mixture of sodium channels with and without associated FHF2, but rather the mixture of models was employed to achieve a closer modeling of voltage dependent inactivation to recorded values, as presented in </span><span style='font-size: 11.0pt'>Online</span><span style='font-size:11.0pt;font-family:Times'> Table VII in Park et al., <span class=SpellE>Circ</span> Res 127, in press, 2020.<span style="mso-spacerun:yes"> </span>It is also important to emphasize that for each sodium channel model, the maximum available sodium conductance upon simulated step depolarization from -135 mV to -30 mV is not equal to </span><!--[if gte msEquation 12]><m:oMath><m:acc><m:accPr><m:chr m:val="̅"/><span style='font-size:11.0pt;mso-ansi-font-size:11.0pt; mso-bidi-font-size:11.0pt;font-family:"Cambria Math";mso-ascii-font-family: "Cambria Math";mso-hansi-font-family:"Cambria Math"'><m:ctrlPr></m:ctrlPr></span></m:accPr><m:e><span style='font-size:11.0pt;font-family:Times'><m:r><span><m:rPr><m:nor/></m:rPr>gNav</m:r></span></span></m:e></m:acc></m:oMath><![endif]--><![if !msEquation]><span style='font-size:12.0pt;font-family:"Times New Roman";mso-fareast-font-family: "Times New Roman";position:relative;top:2.0pt;mso-text-raise:-2.0pt;mso-ansi-language: EN-US;mso-fareast-language:EN-US;mso-bidi-language:AR-SA'><img width=26 height=15 id="_x0000_i1025" src="README_file/image004.png"></span><![endif]><span style='font-size:11.0pt;font-family:Times'>, but is equal to </span><!--[if gte msEquation 12]><m:oMath><m:acc><m:accPr><m:chr m:val="̅"/><span style='font-size:11.0pt;mso-ansi-font-size:11.0pt; mso-bidi-font-size:11.0pt;font-family:"Cambria Math";mso-ascii-font-family: "Cambria Math";mso-hansi-font-family:"Cambria Math"'><m:ctrlPr></m:ctrlPr></span></m:accPr><m:e><span style='font-size:11.0pt;font-family:Times'><m:r><span><m:rPr><m:nor/></m:rPr>gNav</m:r></span></span></m:e></m:acc></m:oMath><![endif]--><![if !msEquation]><span style='font-size:12.0pt;font-family:"Times New Roman";mso-fareast-font-family: "Times New Roman";position:relative;top:2.0pt;mso-text-raise:-2.0pt;mso-ansi-language: EN-US;mso-fareast-language:EN-US;mso-bidi-language:AR-SA'><img width=26 height=15 id="_x0000_i1025" src="README_file/image005.png"></span><![endif]><span style='font-size:11.0pt;font-family:Times'><span style="mso-spacerun:yes"> </span>* (<span class=SpellE>C<sub>off</sub></span>/[C<sub>on</sub>+ <span class=SpellE>C<sub>off</sub></span>]). For the <span class=SpellE>NAV_noF</span> model, <span class=SpellE>C<sub>off</sub></span>/[C<sub>on</sub>+ <span class=SpellE>C<sub>off</sub></span>] = 0.1667, while for the <span class=SpellE>NAV_withF</span> model, <span class=SpellE>C<sub>off</sub></span>/[C<sub>on</sub>+ <span class=SpellE>C<sub>off</sub></span>] = 0.9259.<span style="mso-spacerun:yes"> </span>Since <i style='mso-bidi-font-style: normal'>Fhf2<sup>WT</sup></i> and <i style='mso-bidi-font-style:normal'>Fhf2<sup>KO</sup></i> model <span class=SpellE>cardiomyocytes</span> were tuned to generate the same peak sodium current upon step depolarization from -135 mV, consistent with our recorded cardiomyocyte data, </span><!--[if gte msEquation 12]><m:oMath><m:acc><m:accPr><m:chr m:val="̅"/><span style='font-size:11.0pt;mso-ansi-font-size:11.0pt; mso-bidi-font-size:11.0pt;font-family:"Cambria Math";mso-ascii-font-family: "Cambria Math";mso-hansi-font-family:"Cambria Math"'><m:ctrlPr></m:ctrlPr></span></m:accPr><m:e><span style='font-size:11.0pt;font-family:Times'><m:r><span><m:rPr><m:nor/></m:rPr>gNav</m:r></span></span></m:e></m:acc></m:oMath><![endif]--><![if !msEquation]><span style='font-size:12.0pt;font-family:"Times New Roman";mso-fareast-font-family: "Times New Roman";position:relative;top:2.0pt;mso-text-raise:-2.0pt;mso-ansi-language: EN-US;mso-fareast-language:EN-US;mso-bidi-language:AR-SA'><img width=26 height=15 id="_x0000_i1025" src="README_file/image006.png"></span><![endif]><span style='font-size:11.0pt;font-family:Times'><span style="mso-spacerun:yes"> </span>is greater in the <i style='mso-bidi-font-style: normal'>Fhf2<sup>KO</sup></i> model cardiomyocyte.<span style="mso-spacerun:yes"> </span>As stipulated above, that the maximum available conductance in <span class=SpellE>NAV_noF</span> model is far less than its </span><!--[if gte msEquation 12]><m:oMath><m:acc><m:accPr><m:chr m:val="̅"/><span style='font-size:11.0pt;mso-ansi-font-size:11.0pt; mso-bidi-font-size:11.0pt;font-family:"Cambria Math";mso-ascii-font-family: "Cambria Math";mso-hansi-font-family:"Cambria Math"'><m:ctrlPr></m:ctrlPr></span></m:accPr><m:e><span style='font-size:11.0pt;font-family:Times'><m:r><span><m:rPr><m:nor/></m:rPr>gNav</m:r></span></span></m:e></m:acc></m:oMath><![endif]--><![if !msEquation]><span style='font-size:12.0pt;font-family:"Times New Roman";mso-fareast-font-family: "Times New Roman";position:relative;top:2.0pt;mso-text-raise:-2.0pt;mso-ansi-language: EN-US;mso-fareast-language:EN-US;mso-bidi-language:AR-SA'><img width=26 height=15 id="_x0000_i1025" src="README_file/image007.png"></span><![endif]><span style='font-size:11.0pt;font-family:Times'><span style="mso-spacerun:yes"> </span>is not meant to necessarily imply that most sodium channels in real <i style='mso-bidi-font-style:normal'>Fhf2<sup>KO</sup></i> <span class=SpellE>cardiomyocytes</span> are inactivated under all conditions.<o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph; text-indent:36.0pt'><span style='font-size:11.0pt;font-family:Times'>In the Markov models, </span><span style='font-size:11.0pt;font-family:Symbol'>a</span><span style='font-size:11.0pt;font-family:Times'> and </span><span style='font-size: 11.0pt;font-family:Symbol'>b</span><span style='font-size:11.0pt;font-family: Times'> are <span class=GramE>voltage(</span>v)-dependent rate constants, Q10gate is the <span class=SpellE>thermodyamic</span> scaling factor for rate constants, and Q10cond is the thermodynamic scaling factor for conductance. Parameters with equivalent values for <span class=SpellE>NAV_withF</span> and <span class=SpellE>NAV_noF</span> models are: n1 = 100, n2 = n3 = 20, n4 = 3, n5 = 1.5, n6 = 0.75, ‘</span><span style='font-size:11.0pt;font-family:Symbol'>a</span><span style='font-size:11.0pt;font-family:Times'>‘ = 2.44375 (<i style='mso-bidi-font-style: normal'>ms<sup>-1</sup></i>), ‘</span><span style='font-size:11.0pt;font-family: Symbol'>b</span><span style='font-size:11.0pt;font-family:Times'>‘ = 0.01325 (<i style='mso-bidi-font-style:normal'>ms<sup>-1</sup></i>), V</span><span style='font-size:11.0pt;font-family:Symbol'>a</span><span style='font-size: 11.0pt;font-family:Times'> = V</span><span style='font-size:11.0pt;font-family: Symbol'>b</span><span style='font-size:11.0pt;font-family:Times'> = 9 (<i style='mso-bidi-font-style:normal'>mV</i>), </span><span style='font-size:11.0pt; font-family:Symbol'>g</span><span style='font-size:11.0pt;font-family:Times'> = 150 (<i style='mso-bidi-font-style:normal'>ms<sup>-1</sup></i>), </span><span style='font-size:11.0pt;font-family:Symbol'>d</span><span style='font-size: 11.0pt;font-family:Times'> = 40 (<i style='mso-bidi-font-style:normal'>ms<sup>-1</sup></i>), ‘<span class=SpellE>O’<sub>off</sub></span> = 0.0005 (<i style='mso-bidi-font-style: normal'>ms<sup>-1</sup></i>).<span style="mso-spacerun:yes"> </span>Parameters with different values for <span class=SpellE>NAV_withF</span> <span class=SpellE>vs</span> <span class=SpellE>NAV_noF</span> models are: ‘<span class=SpellE>C’<sub>on</sub></span> = 0.004 <span class=SpellE><i>vs</i></span> 0.025 (<i style='mso-bidi-font-style:normal'>ms<sup>-1</sup></i>), ‘<span class=SpellE>C’<sub>off</sub></span> = 0.05 <span class=SpellE><i>vs</i></span> 0.005 (<i style='mso-bidi-font-style:normal'>ms<sup>-1</sup></i>), ‘<span class=SpellE>O’<sub>on</sub></span> = 0.85 <span class=SpellE><i style='mso-bidi-font-style:normal'>vs</i></span> 1.3 (<i style='mso-bidi-font-style: normal'>ms<sup>-1</sup></i>), <span class=SpellE>V<sub>shift</sub></span> = -54 <span class=SpellE><i style='mso-bidi-font-style:normal'>vs</i></span> -57.5<i style='mso-bidi-font-style:normal'> </i>(<i style='mso-bidi-font-style:normal'>mV</i>).<span style="mso-spacerun:yes"> </span>In the <i style='mso-bidi-font-style: normal'>Fhf2<sup>KO</sup></i> cardiomyocyte, </span><!--[if gte msEquation 12]><m:oMath><m:acc><m:accPr><m:chr m:val="̅"/><span style='font-size:11.0pt;mso-ansi-font-size:11.0pt; mso-bidi-font-size:11.0pt;font-family:"Cambria Math";mso-ascii-font-family: "Cambria Math";mso-hansi-font-family:"Cambria Math"'><m:ctrlPr></m:ctrlPr></span></m:accPr><m:e><span style='font-size:11.0pt;font-family:Times'><m:r><span><m:rPr><m:nor/></m:rPr>gNav</m:r></span></span></m:e></m:acc></m:oMath><![endif]--><![if !msEquation]><span style='font-size:12.0pt;font-family:"Times New Roman";mso-fareast-font-family: "Times New Roman";position:relative;top:2.0pt;mso-text-raise:-2.0pt;mso-ansi-language: EN-US;mso-fareast-language:EN-US;mso-bidi-language:AR-SA'><img width=26 height=15 id="_x0000_i1025" src="README_file/image008.png"></span><![endif]><span style='font-size:11.0pt;font-family:Times'>_<span class=SpellE>noF</span><i style='mso-bidi-font-style:normal'> </i>= 25 <span class=SpellE>nS</span>/pF, while in the <i style='mso-bidi-font-style:normal'>Fhf2<sup>WT</sup></i> cardiomyocyte, </span><!--[if gte msEquation 12]><m:oMath><m:acc><m:accPr><m:chr m:val="̅"/><span style='font-size:11.0pt;mso-ansi-font-size:11.0pt; mso-bidi-font-size:11.0pt;font-family:"Cambria Math";mso-ascii-font-family: "Cambria Math";mso-hansi-font-family:"Cambria Math"'><m:ctrlPr></m:ctrlPr></span></m:accPr><m:e><span style='font-size:11.0pt;font-family:Times'><m:r><span><m:rPr><m:nor/></m:rPr>gNav</m:r></span></span></m:e></m:acc></m:oMath><![endif]--><![if !msEquation]><span style='font-size:12.0pt;font-family:"Times New Roman";mso-fareast-font-family: "Times New Roman";position:relative;top:2.0pt;mso-text-raise:-2.0pt;mso-ansi-language: EN-US;mso-fareast-language:EN-US;mso-bidi-language:AR-SA'><img width=26 height=15 id="_x0000_i1025" src="README_file/image009.png"></span><![endif]><span style='font-size:11.0pt;font-family:Times'>_<span class=SpellE>noF</span> = 8.83 <span class=SpellE>nS</span>/pF and </span><!--[if gte msEquation 12]><m:oMath><m:acc><m:accPr><m:chr m:val="̅"/><span style='font-size:11.0pt;mso-ansi-font-size:11.0pt; mso-bidi-font-size:11.0pt;font-family:"Cambria Math";mso-ascii-font-family: "Cambria Math";mso-hansi-font-family:"Cambria Math"'><m:ctrlPr></m:ctrlPr></span></m:accPr><m:e><span style='font-size:11.0pt;font-family:Times'><m:r><span><m:rPr><m:nor/></m:rPr>gNav</m:r></span></span></m:e></m:acc></m:oMath><![endif]--><![if !msEquation]><span style='font-size:12.0pt;font-family:"Times New Roman";mso-fareast-font-family: "Times New Roman";position:relative;top:2.0pt;mso-text-raise:-2.0pt;mso-ansi-language: EN-US;mso-fareast-language:EN-US;mso-bidi-language:AR-SA'><img width=26 height=15 id="_x0000_i1025" src="README_file/image010.png"></span><![endif]><span style='font-size:11.0pt;font-family:Times'>_<span class=SpellE>withF</span><span style="mso-spacerun:yes"> </span>= 2.94 <span class=SpellE>nS</span>/<span class=SpellE>pF.</span><span style="mso-spacerun:yes"> </span><o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph; text-indent:36.0pt'><span style='font-size:11.0pt'>Online Table VII </span><span style='font-size:11.0pt;font-family:Times'>in Park et al., <i style='mso-bidi-font-style: normal'>Circ. Res.</i> 127, in press, 2020</span><span style='font-size:11.0pt'> presents the <span class=SpellE>Na<sub>V</sub></span> inactivation and activation characteristics and generated currents of the <i style='mso-bidi-font-style: normal'>Fhf2<sup>WT</sup></i> and <i style='mso-bidi-font-style:normal'>Fhf2<sup>KO </sup></i>cardiomyocyte models, which are in close agreement with sodium current recordings from <i style='mso-bidi-font-style:normal'>Fhf2<sup>WT</sup></i> and <i style='mso-bidi-font-style:normal'>Fhf2<sup>KO</sup></i> ventricular <span class=SpellE>cardiomyocytes</span> (Park et al, <i style='mso-bidi-font-style: normal'>Nature Comm.</i> 7:12966, 2016; Wang et al, <i style='mso-bidi-font-style: normal'>J. Mol. Cell. <span class=SpellE>Cardiol</span>. </i>104:63, 2017; Park et al, Circ. Res. </span><i style='mso-bidi-font-style:normal'><span style='font-size:11.0pt;font-family:Times'>Circ. Res.</span></i><span style='font-size:11.0pt;font-family:Times'> 127, in press, 2020)</span><span style='font-size:11.0pt'>.<span style="mso-spacerun:yes"> </span></span><span style='font-size:11.0pt;font-family:Times'>The </span><!--[if gte msEquation 12]><m:oMath><m:acc><m:accPr><m:chr m:val="̅"/><span style='font-size:11.0pt;mso-ansi-font-size:11.0pt; mso-bidi-font-size:11.0pt;font-family:"Cambria Math";mso-ascii-font-family: "Cambria Math";mso-hansi-font-family:"Cambria Math"'><m:ctrlPr></m:ctrlPr></span></m:accPr><m:e><span style='font-size:11.0pt;font-family:Times'><m:r><span><m:rPr><m:nor/></m:rPr>gNav</m:r></span></span></m:e></m:acc></m:oMath><![endif]--><![if !msEquation]><span style='font-size:12.0pt;font-family:"Times New Roman";mso-fareast-font-family: "Times New Roman";position:relative;top:2.0pt;mso-text-raise:-2.0pt;mso-ansi-language: EN-US;mso-fareast-language:EN-US;mso-bidi-language:AR-SA'><img width=26 height=15 id="_x0000_i1025" src="README_file/image011.png"></span><![endif]><span style='font-size:11.0pt;font-family:Times'><span style="mso-spacerun:yes"> </span>densities for the <i style='mso-bidi-font-style: normal'>Fhf2<sup>WT</sup></i> cardiomyocyte were estimated to generate action potential amplitude in isolated cardiomyocyte model with amplitude similar to prior recordings and conduction velocity in model strand comparable to velocity reported by optical mapping, while </span><!--[if gte msEquation 12]><m:oMath><m:acc><m:accPr><m:chr m:val="̅"/><span style='font-size:11.0pt;mso-ansi-font-size:11.0pt; mso-bidi-font-size:11.0pt;font-family:"Cambria Math";mso-ascii-font-family: "Cambria Math";mso-hansi-font-family:"Cambria Math"'><m:ctrlPr></m:ctrlPr></span></m:accPr><m:e><span style='font-size:11.0pt;font-family:Times'><m:r><span><m:rPr><m:nor/></m:rPr>gNav</m:r></span></span></m:e></m:acc></m:oMath><![endif]--><![if !msEquation]><span style='font-size:12.0pt;font-family:"Times New Roman";mso-fareast-font-family: "Times New Roman";position:relative;top:2.0pt;mso-text-raise:-2.0pt;mso-ansi-language: EN-US;mso-fareast-language:EN-US;mso-bidi-language:AR-SA'><img width=26 height=15 id="_x0000_i1025" src="README_file/image012.png"></span><![endif]><span style='font-size:11.0pt;font-family:Times'><span style="mso-spacerun:yes"> </span>for <span class=SpellE>NAV_noF</span> in <i style='mso-bidi-font-style:normal'>Fhf2<sup>KO</sup></i> cells allowed <i style='mso-bidi-font-style:normal'>Fhf2<sup>WT</sup></i> and <i style='mso-bidi-font-style:normal'>Fhf2<sup>KO</sup></i> model <span class=SpellE>cardiomyocytes</span> to generate same peak sodium current upon depolarization from -135 mV holding potential, as previously demonstrated empirically</span><!--[if supportFields]><span style='font-size:11.0pt; font-family:Times'><span style='mso-element:field-begin'></span> ADDIN EN.CITE <span style='mso-element:field-begin'></span><span style="mso-spacerun:yes"> </span>ADDIN EN.CITE.DATA <![if gte mso 9]><xml> 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</xml><![endif]--></span><!--[if supportFields]><span style='font-size:11.0pt; font-family:Times'><span style='mso-element:field-end'></span></span><![endif]--><span style='font-size:11.0pt;font-family:Times'>.<span style="mso-spacerun:yes"> </span></span><span style='font-size:11.0pt'>A third cardiomyocyte model termed <i style='mso-bidi-font-style:normal'>Fhf2<sup>WT</sup>Na<sub>V</sub><sup>HYPO</sup></i> has the same <span class=SpellE>Na<sub>V</sub></span> gating parameters as the <i style='mso-bidi-font-style:normal'>Fhf2<sup>WT</sup></i> model, but the <span class=SpellE>Nav</span> densities are reduced by a factor of 0.49 so that the <i style='mso-bidi-font-style:normal'>Fhf2<sup>WT</sup>Na<sub>V</sub><sup>HYPO </sup></i>and <i style='mso-bidi-font-style:normal'>Fhf2<sup>KO</sup></i> models generate the same I-<span class=SpellE>Na<sub>peak</sub></span> when depolarized from a -87mV <span style='color:black;mso-themecolor:text1'>resting </span>potential (Online Table VII </span><span style='font-size:11.0pt;font-family:Times'>in Park et al., <i style='mso-bidi-font-style:normal'>Circ. Res.</i> 127, in press, 2020</span><span style='font-size:11.0pt'>).<span style="mso-spacerun:yes"> </span></span><span style='font-size:11.0pt;font-family:Times'><o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph; text-indent:36.0pt'><span style='font-size:11.0pt;font-family:Times'>Nomenclature clarification:<span style="mso-spacerun:yes"> </span>The names of rate parameters with units <i style='mso-bidi-font-style:normal'>ms<sup>-1</sup></i> above that are flanked by apostrophes in Park et al., <i style='mso-bidi-font-style: normal'>Circ. Res.</i> 127, in press, 20 are named differently in the uploaded <span class=SpellE>Nav</span> models, where the rate parameters are instead preceded by the prefix A.<span style="mso-spacerun:yes"> </span>As examples, ‘</span><span style='font-size:11.0pt;font-family:Symbol'>a</span><span style='font-size: 11.0pt;font-family:Times'>‘ in the publication is equivalent to A</span><span style='font-size:11.0pt;font-family:Symbol'>a</span><span style='font-size: 11.0pt;font-family:Times'> in the uploaded model, ‘<span class=SpellE>C’<sub>off</sub></span> is equivalent to <span class=SpellE>AC<sub>off</sub></span>, etc.<span style="mso-spacerun:yes"> </span>Additionally, </span><!--[if gte msEquation 12]><m:oMath><m:acc><m:accPr><m:chr m:val="̅"/><span style='font-size:11.0pt;mso-ansi-font-size:11.0pt; mso-bidi-font-size:11.0pt;font-family:"Cambria Math";mso-ascii-font-family: "Cambria Math";mso-hansi-font-family:"Cambria Math"'><m:ctrlPr></m:ctrlPr></span></m:accPr><m:e><span style='font-size:11.0pt;font-family:Times'><m:r><span><m:rPr><m:nor/></m:rPr>gNav</m:r></span></span></m:e></m:acc></m:oMath><![endif]--><![if !msEquation]><span style='font-size:12.0pt;font-family:"Times New Roman";mso-fareast-font-family: "Times New Roman";position:relative;top:2.0pt;mso-text-raise:-2.0pt;mso-ansi-language: EN-US;mso-fareast-language:EN-US;mso-bidi-language:AR-SA'><img width=26 height=15 id="_x0000_i1025" src="README_file/image013.png"></span><![endif]><span style='font-size:11.0pt;font-family:Times'><span style="mso-spacerun:yes"> </span>in the publication is equivalent to <span class=SpellE>gnabar</span> in the uploaded model.<o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph; text-indent:36.0pt'><span style='font-size:11.0pt;font-family:Times'><o:p> </o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><u><span style='font-size:11.0pt;font-family:Times'>Voltage-gated calcium conductance:</span></u><span style='font-size:11.0pt;font-family:Times'><span style="mso-spacerun:yes"> </span><i style='mso-bidi-font-style:normal'>Fhf2<sup>WT</sup></i> and <i style='mso-bidi-font-style:normal'>Fhf2<sup>KO</sup></i> cardiomyocyte models now have an equivalent L-type voltage-gated calcium conductance expressed through an 8-state Markov model (Scheme 2) based upon the equivalent calcium current density, voltage dependence of activation and steady-state inactivation, and voltage-dependent rate of inactivation measured empirically in <i style='mso-bidi-font-style:normal'>Fhf2<sup>WT</sup></i> and <i style='mso-bidi-font-style:normal'>Fhf2<sup>KO</sup></i> <span class=SpellE>cardiomyocytes</span> (Figure 2 and Table 1 in Park et al., <i style='mso-bidi-font-style:normal'>Circ. Res.</i> 127, in press, 20).<span style="mso-spacerun:yes"> </span><o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><u><span style='font-size:11.0pt;font-family:Times'>Scheme 2<o:p></o:p></span></u></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span lang=IT style='font-size:11.0pt;font-family:Times;mso-ansi-language:IT; mso-fareast-language:IT;mso-no-proof:yes'><!--[if gte vml 1]><v:shape id="Picture_x0020_2" o:spid="_x0000_i1025" type="#_x0000_t75" style='width:245pt;height:106pt; visibility:visible;mso-wrap-style:square'> <v:imagedata src="README_file/image014.png" o:title=""/> </v:shape><![endif]--><![if !vml]><img width=247 height=108 src="README_file/image015.png" v:shapes="Picture_x0020_2"><![endif]></span><span style='font-size:11.0pt;font-family:Times'><o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times'>The kinetic parameters are: Q10 = 3<sup>{(<span style='position:relative;top:-3.0pt;mso-text-raise:3.0pt'>o</span>C – 32.76)/10}</sup>, </span><span style='font-size:11.0pt;font-family:Symbol'>a</span><span style='font-size:11.0pt;font-family:Times'> = Q10 * 11.74 * 10<sup>{(V<span style='position:relative;top:3.0pt;mso-text-raise:-3.0pt'>m</span><span style="mso-spacerun:yes"> </span>+ 17)/50}</sup> <i style='mso-bidi-font-style: normal'>(ms<sup>-1</sup>)</i>, </span><span style='font-size:11.0pt;font-family: Symbol'>b</span><span style='font-size:11.0pt;font-family:Times'> = Q10 * 0.0324 * 10<sup>{(-V<span style='position:relative;top:3.0pt;mso-text-raise: -3.0pt'>m</span><span style="mso-spacerun:yes"> </span>- 17)/5.5}</sup> <i style='mso-bidi-font-style:normal'>(ms<sup>-1</sup>)</i>, <i style='mso-bidi-font-style: normal'>n<sub>1</sub></i> = 32.532, <i style='mso-bidi-font-style:normal'>n<sub>2</sub></i> = 0.123, </span><span style='font-size:11.0pt;font-family:Symbol'>g</span><span style='font-size:11.0pt;font-family:Times'> = Q10 * 150 <i style='mso-bidi-font-style: normal'>(ms<sup>-1</sup>)</i>, </span><span style='font-size:11.0pt;font-family: Symbol'>d</span><span style='font-size:11.0pt;font-family:Times'> = Q10 * 40 <i style='mso-bidi-font-style:normal'>(ms<sup>-1</sup>)</i>, <i style='mso-bidi-font-style: normal'>C<sub>on</sub></i> = Q10 * 0.001 <i style='mso-bidi-font-style:normal'>(ms<sup>-1</sup>)</i>, <span class=SpellE><i style='mso-bidi-font-style:normal'>C<sub>off</sub></i></span> = Q10 * 10 <i style='mso-bidi-font-style:normal'>(ms<sup>-1</sup>)</i>, <span class=SpellE><i style='mso-bidi-font-style:normal'>O<sub>on</sub></i></span> = Q10 * 0.2 <i style='mso-bidi-font-style:normal'>(ms<sup>-1</sup>)</i>, <span class=SpellE><i style='mso-bidi-font-style:normal'>O<sub>off</sub></i></span> = Q10 * 0.001 <i style='mso-bidi-font-style:normal'>(ms<sup>-1</sup>)</i>, a = (<span class=SpellE><i style='mso-bidi-font-style:normal'>O<sub>on</sub></i></span>/<span class=GramE><i style='mso-bidi-font-style:normal'>C<sub>on</sub></i>)<sup>0.5</sup></span>, b = (<span class=SpellE><i style='mso-bidi-font-style:normal'>O<sub>off</sub></i></span>/<span class=SpellE><i style='mso-bidi-font-style:normal'>C<sub>off</sub></i></span>)<sup>0.5</sup>, where <span class=SpellE>V<sub>m</sub></span> is membrane voltage.<o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt'><o:p> </o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><u><span style='font-size:11.0pt;font-family:Times'>Potassium <span class=SpellE>conductances</span>:</span></u><span style='font-size:11.0pt;font-family:Times'><span style="mso-spacerun:yes"> </span>The potassium <span class=SpellE>conductances</span> are taken from <span class=SpellE>Bondarenko</span> et al., </span><i style='mso-bidi-font-style:normal'><span style='mso-no-proof:yes'>Am. J. Physiol. Heart. Circ. Physiol</span></i><span style='mso-no-proof:yes'>. 287:H1378, 2004,</span><span style='font-size:11.0pt;font-family:Times'> and include the time-dependent <span class=SpellE>conductances</span> fast transient outward conductance (<span class=SpellE>g_Kto_f</span>), <span class=SpellE>noninactivating</span> <span class=SpellE>ultrarapid</span> delayed <span class=SpellE>rectifer</span> (<span class=SpellE>g_Kurdr</span>), <span class=SpellE>noninactivating</span> rapid delayed rectifier (<span class=SpellE>g_Krdr</span>), <span class=SpellE>noninactivating</span> slow delayed rectifier (<span class=SpellE>g_Ksdr</span>), and steady-state conductance (<span class=SpellE>g_Kss</span>), along with time-independent conductance (<span class=SpellE>g_Kti</span>) that has both leak and inward rectifier components.<span style="mso-spacerun:yes"> </span>The weights of these <span class=SpellE>conductances</span> were adjusted 1) to give passive property </span><span style='font-size:11.0pt;font-family:Symbol'>D</span><span style='font-size:11.0pt;font-family:Times'>V as function of injected current similar to empirically recorded dissociated ventricular <span class=SpellE>cardiomyocytes</span> (</span><span style='font-size:11.0pt'>Park et al, <i style='mso-bidi-font-style: normal'>Nature Comm.</i> 7:12966, 2016</span><span style='font-size:11.0pt; font-family:Times'>), and 2) to give a decay in the action potential in cardiomyocyte strand models similar to measured action potential decay optically recorded in paced ventricular myocardium (Online Fig II in Park et al., <i style='mso-bidi-font-style:normal'>Circ. Res.</i> 127, in press, 2020).<span style="mso-spacerun:yes"> </span>These conductance values (S/</span><span style='font-size:11.0pt;font-family:Symbol'>m</span><span style='font-size: 11.0pt;font-family:Times'>F) are <span class=SpellE>g_Kti</span> = 0.00021, <span class=SpellE>g_Kss</span> <span class=GramE>=<span style="mso-spacerun:yes"> </span>0.00007</span>, <span class=SpellE>g_Kto_f</span> = 0.0000235, <span class=SpellE>g_Kurdr</span> = 0.000025, <span class=SpellE>g_Krdr</span> = 0.000468, <span class=SpellE>g_Ksdr</span> = 0.00000575.<o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times'><o:p> </o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><u><span style='font-size:11.0pt;font-family:Times'>Other time-independent currents:</span></u><span style='font-size:11.0pt;font-family:Times'><span style="mso-spacerun:yes"> </span>Two other small currents were incorporated to maintain <span class=SpellE>cardiomyocytes</span> at -87 mV resting potential at all temperatures.<span style="mso-spacerun:yes"> </span>Background sodium conductance (<span class=SpellE>g_Nabg</span> = 0.0000018 S/</span><span style='font-size:11.0pt;font-family:Symbol'>m</span><span style='font-size:11.0pt;font-family:Times'>F) is taken from <span class=SpellE>Bondarenko</span> et al., </span><i style='mso-bidi-font-style:normal'><span style='mso-no-proof: yes'>Am. J. Physiol. Heart. Circ. Physiol</span></i><span style='mso-no-proof: yes'>. 287:H1378, 2004</span><span style='font-size:11.0pt;font-family:Times'>, while a temperature-dependent nonspecific current (<span class=SpellE>i_ITEMP</span>) was incorporated to offset small temperature variations in <span class=SpellE>conductances</span> near the resting potential, set at <span class=SpellE>i_ITEMP</span> = 0.0322 * {43 – (<span class=SpellE><sup>o</sup>C</span>)} (<span class=SpellE>pA</span>/pF).<o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><b style='mso-bidi-font-weight:normal'><span style='font-size:11.0pt'><o:p> </o:p></span></b></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><b style='mso-bidi-font-weight:normal'><i style='mso-bidi-font-style:normal'><span style='font-size:11.0pt'>Cardiomyocyte strand simulations.</span></i></b><span style='font-size:11.0pt'> The cardiomyocyte strands have a resting membrane potential of -87 mV.<span style="mso-spacerun:yes"> </span>In all simulations, a 100 <span class=SpellE>msec</span> delay was employed to allow the <span class=SpellE>Na<sub>V</sub></span> Markov models to reach steady state prior to injecting the first cell with two 0.5 <span class=SpellE>msec</span> current stimuli at 10 Hz.<span style="mso-spacerun:yes"> </span>Injected current amplitude was adjusted to achieve maximal induced sodium current in the first cell.<span style="mso-spacerun:yes"> </span>The generated currents and voltages throughout the strand were analyzed only following the second stimulus, which takes into account the states of dynamic <span class=SpellE>conductances</span> at sinus rhythm<span style='color:black;mso-themecolor:text1'>. <span style='background:white'>All simulations were conducted using <span class=SpellE>Cvode</span> multi-order variable time step integration method.</span> </span>For each model, simulations were run after either elevating temperature in 1<sup>o</sup>C increments or reducing <span class=SpellE>junctional</span>, sodium, or calcium <span class=SpellE>conductances</span> in 1% increments.<span style="mso-spacerun:yes"> </span>Conduction safety was defined as propagation of action potentials through the entire strand, with regenerative sodium current reaching steady cell-to-cell amplitude.<span style="mso-spacerun:yes"> </span>Conduction failure was defined as a failure to generate sodium current in all cells throughout the strand with accompanying fall-off in depolarization amplitudes.<span style="mso-spacerun:yes"> </span>In most simulations, conductance parameters were altered equivalently in all cells within the strand.<span style="mso-spacerun:yes"> </span>However, to investigate how calcium conductance contributes to conduction safety in the <i style='mso-bidi-font-style: normal'>Fhf2<sup>KO</sup></i> strand, we conducted simulations where calcium conductance was deleted from cells 51-111 only.<o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt'><span style='mso-tab-count:1'> </span><o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt'>Under any simulation condition, the <i style='mso-bidi-font-style:normal'>Fhf2<sup>KO</sup></i> <span class=SpellE>cardiomyocytes</span> generate substantially less sodium current than <i style='mso-bidi-font-style: normal'>Fhf2<sup>WT</sup></i> <span class=SpellE>cardiomyocytes</span> for two reasons: 1) at resting potential, approximately 74% of the sodium conductance is inactivated in <i style='mso-bidi-font-style:normal'>Fhf2<sup>KO</sup></i> cells, while there is only ~50% inactivation of the sodium conductance in <i style='mso-bidi-font-style:normal'>Fhf2<sup>WT</sup></i> cells (Figure 4E,F, Online Table VII </span><span style='font-size:11.0pt;font-family:Times'>in Park et al., <i style='mso-bidi-font-style:normal'>Circ. Res.</i> 127, in press, 2020</span><span style='font-size:11.0pt'>), and 2) the <span class=SpellE>Na<sub>V</sub></span> model in <i style='mso-bidi-font-style:normal'>Fhf2<sup>KO</sup></i> <span class=SpellE>cardiomyocytes</span> has faster rates of closed-state and open-state inactivation than does the <span class=SpellE>Na<sub>v</sub></span> model in <i style='mso-bidi-font-style:normal'>Fhf2<sup>WT</sup></i> cells (Online Table VII </span><span style='font-size:11.0pt;font-family:Times'>in Park et al., <i style='mso-bidi-font-style:normal'>Circ. Res.</i> 127, in press, 2020</span><span style='font-size:11.0pt'>).<span style="mso-spacerun:yes"> </span>Action potential amplitudes, conduction velocity, [<span class=SpellE>dV</span>/<span class=SpellE>dt</span>]<sub><span style='color:black;mso-themecolor:text1'>max</span></sub>, safety factor (SF), and conduction safety or failure thresholds in the <i style='mso-bidi-font-style:normal'>Fhf2<sup>WT</sup></i>and <i style='mso-bidi-font-style:normal'>Fhf2<sup>KO</sup></i> strands in response to variations of temperature, <span class=SpellE>Gj</span>, <span class=SpellE>gNa<sub>V</sub></span>, or <span class=SpellE>gCa<sub>V</sub></span> are summarized in Online Table VI </span><span style='font-size:11.0pt;font-family:Times'>in Park et al., <i style='mso-bidi-font-style: normal'>Circ. Res.</i> 127, in press, 2020</span><span style='font-size:11.0pt'>.<span style="mso-spacerun:yes"> </span>SF values were calculated based upon its originally described formulation (Shaw and Rudy, <i style='mso-bidi-font-style: normal'>Circ. Res.</i> 81:727, 1997), except that all membrane currents (sodium, calcium, potassium, capacitive) were incorporated into the calculation for determining when a cell transitioned from being predominantly a sink to a source.<o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times'><o:p> </o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><b style='mso-bidi-font-weight:normal'><span style='font-size:11.0pt;font-family: Times'>USE OF THE MODELS<o:p></o:p></span></b></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><b style='mso-bidi-font-weight:normal'><i style='mso-bidi-font-style:normal'><span style='font-size:11.0pt;font-family:Times'><o:p> </o:p></span></i></b></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times'>1) Download the Ventricular_GUI.zip file of the model.<span style="mso-spacerun:yes"> </span>Extract the embedded files.<o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times'>2) Open NEURON.<span style="mso-spacerun:yes"> </span>Run the <span class=SpellE>mknrn</span> program, and use it to select the <span class=SpellE>Ventricular_GUI</span> folder and convert the .mod files into compiled .o files.<o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times'>3) Launch the model by double-clicking Start_GUI_3.<span style="mso-spacerun:yes"> </span>This will open the neuron.exe terminal and several GUI windows:<span style="mso-spacerun:yes"> </span><q>Main Menu</q>, <q>Set model <span class=SpellE>paramters</span></q>, and <q>Set <span class=SpellE>Sim</span> Structure</q>.<o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times'><o:p> </o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times'>The <q>Set <span class=SpellE>Sim</span> Structure</q> window allows for the selection of the <i style='mso-bidi-font-style: normal'><span style='color:black;mso-themecolor:text1'>Fhf2<sup>WT</sup> </span></i><span style='color:black;mso-themecolor:text1'>or <i style='mso-bidi-font-style:normal'>Fhf2<sup>KO</sup> </i>cardiomyocyte models, the temperature (37<sup>o</sup>C default), the number of <span class=SpellE>myocytes</span> in the strand (default 111), and the gap <span class=SpellE>junctional</span> conductance between cells along the strand (default 772,800 <span class=SpellE>pS</span>).<span style="mso-spacerun:yes"> </span>Toggling between the WT and KO models using the<span style="mso-spacerun:yes"> </span><q>Activate KO Mutation</q> button alters the conductance densities for the <span class=SpellE>Nav_withF</span> and <span class=SpellE>Nav_noF</span> sodium channel models, and these densities are seen in the <q>Set Model Parameters</q> window.<span style="mso-spacerun:yes"> </span>The selected strand model can be launched from the button <q>Linear Propagation Along Strand</q>, which generates additional windows, including stimulus electrodes positioned within the first cell <span class=SpellE><span class=GramE>myocyte.o</span></span><span class=GramE>[</span>0] preset to generate 0.5 millisecond pulses of current at 100 <span class=SpellE>msec</span> and 200 <span class=SpellE>msec</span> after simulation initiation.<span style="mso-spacerun:yes"> </span>The <q>Propagation Protocol</q> window allows selection of voltage vs. time, sodium current vs. time, and sodium channel states vs. time, and the simulation is initiated from the Run button.<o:p></o:p></span></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times;color:black;mso-themecolor:text1'><o:p> </o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times;color:black;mso-themecolor:text1'>The <q>Set <span class=SpellE>Sim</span> Structure</q> window has other buttons as well.<span style="mso-spacerun:yes"> </span>For user convenience, other buttons in the <q>Set <span class=SpellE>Sim</span> Structure</q> window launch action potential propagation simulations present in Figures 4B, 4C, 5A, 5B, 5F, 5G, 6A, 6B, 6D, or 6E from </span><span style='font-size:11.0pt;font-family: Times'>Park et al., <i style='mso-bidi-font-style:normal'>Circ. Res.</i> 127, in press, 2020.<span style="mso-spacerun:yes"> </span>These Figure Panel buttons each open a figure panel window and a voltage vs. time graph, and the simulation can be initiated from the Run button within the figure panel window.<span style="mso-spacerun:yes"> </span><i style='mso-bidi-font-style: normal'>It is highly recommended that when wishing to open different configuration buttons from the <span style='color:black;mso-themecolor:text1'><q>Set <span class=SpellE>Sim</span> Structure</q> window, the model should be fully closed by closing the NEURON terminal window, and then <span class=SpellE>relaunching</span> the model from Start_GUI_3.</span></i><span style="mso-spacerun:yes"> </span><o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times;color:black;mso-themecolor:text1'><o:p> </o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times;color:black;mso-themecolor:text1'>In order to facilitate sodium current voltage clamp protocols, the <q></span><span style='font-size:11.0pt;font-family:Times'>Set <span class=SpellE>Sim</span> Structure<span style='color:black;mso-themecolor:text1'></q> window also has buttons to select for protocols to assay <span class=SpellE>Na<sub>v</sub></span> voltage dependence of activation, voltage dependence of steady-state inactivation, and sodium currents in response to variable-rate voltage ramps.<span style="mso-spacerun:yes"> </span>The launch of any of these protocols creates a single cardiomyocyte with the selected genotype and temperature parameters, along with a protocol control window, from which the simulation is initiated with the Run button.<o:p></o:p></span></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times;color:black;mso-themecolor:text1'><o:p> </o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times;color:black;mso-themecolor:text1'>The densities of all ionic <span class=SpellE>conductances</span> can be changed equivalently in all cells of the strand using the <q>Set Model Parameters</q> window.<span style="mso-spacerun:yes"> </span>This window can also be used to modify kinetic parameters for the sodium channel models.<span style="mso-spacerun:yes"> </span>Changes to parameters of ionic and <span class=SpellE>junctional</span> <span class=SpellE>conductances</span> in a subset of cells within the strand can be made through hoc code commands in the terminal window.<span style="mso-spacerun:yes"> </span>As examples, <o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times;color:black;mso-themecolor:text1'>1) The command:<o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times;color:black;mso-themecolor:text1'><span style="mso-spacerun:yes"> </span><span style="mso-spacerun:yes"> </span><span class=GramE>for</span> <span class=SpellE>i</span>=50,110 {<span class=SpellE>prop_myo.myocytes.o</span>[<span class=SpellE>i</span>].<span class=SpellE>cell.gcabar_Ca_L</span> = 0}<o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span class=GramE><span style='font-size:11.0pt;font-family:Times;color:black; mso-themecolor:text1'>sets</span></span><span style='font-size:11.0pt; font-family:Times;color:black;mso-themecolor:text1'> the calcium conductance to zero in cells 51-111 of the strand (note that first cell in model is <span class=SpellE>myocytes.o</span>[0])<o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times;color:black;mso-themecolor:text1'>2) The pair of commands:<o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times;color:black;mso-themecolor:text1'><span style="mso-spacerun:yes"> </span><span class=SpellE><span class=GramE>prop</span>_myo.gap_sources.o</span>[10].g = 10000<o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span style='font-size:11.0pt;font-family:Times;color:black;mso-themecolor:text1'><span style="mso-spacerun:yes"> </span><span class=SpellE><span class=GramE>prop</span>_myo.gap_dests.o</span>[10].g = 10000<o:p></o:p></span></p> <p class=MsoNormal style='text-align:justify;text-justify:inter-ideograph'><span class=GramE><span style='font-size:11.0pt;font-family:Times;color:black; mso-themecolor:text1'>resets</span></span><span style='font-size:11.0pt; font-family:Times;color:black;mso-themecolor:text1'> the gap <span class=SpellE>junctional</span> conductance between cells 11 and 12 in the strand to 10000 <span class=SpellE>pS.</span><o:p></o:p></span></p> <p class=MsoNormal><o:p> </o:p></p> </div> <p><h3>Changelog</h3> 2022-12: Fix upcoming 9.0.0 error: <var> used as both variable and function in files</p> </body> </html>
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FHF2KO and Wild-Type Mouse Cardiomyocyte Strands (Park et al 2020)