Thomas Eschenhagen, Roberto Bolli, Thomas Braun, Loren J. Field, Bernd K. Fleischmann, Jonas Frisén, Mauro Giacca, Joshua M. Hare, Steven R. Houser, Richard T. Lee, Eduardo Marbán, James F. Martin, Jeffery D. Molkentin, Charles E. Murry, Paul R. Riley, Pilar Ruiz-Lozano, Hesham A. Sadek, Mark A. Sussman and Joseph A. Hill
Circulation. 2017;CIRCULATIONAHA.117.029343, originally published July 6, 2017
Michael Didié, Peter Christalla, Alexander P Schwoerer, Heimo Ehmke, John A Scherschel, Michael Rubart, Mark H Soonpaa, Loren J Field, Thomas Eschenhagen and Wolfram H Zimmermann
Ralph Knöll, Wolfgang A. Linke, Peijian Zou, Snježana Miočiċ, Sawa Kostin, Byambajav Buyandelger, Ching-Hsin Ku, Stefan Neef, Monika Bug, Katrin Schäfer, Gudrun Knöll, Leanne E. Felkin, Johannes Wessels, Karl Toischer, Franz Hagn, Horst Kessler, Michael Didié, Thomas Quentin, Lars S. Maier, Nils Teucher, Bernhard Unsöld, Albrecht Schmidt, Emma J. Birks, Sylvia Gunkel, Patrick Lang, Henk Granzier, Wolfram-Hubertus Zimmermann, Loren J. Field, Georgine Faulkner, Matthias Dobbelstein, Paul J.R. Barton, Michael Sattler, Matthias Wilmanns and Kenneth R. Chien
Circulation Research. 2011;109:758-769, originally published September 15, 2011
Ralph Knöll, Wolfgang A. Linke, Peijian Zou, Snježana Miočiċ, Sawa Kostin, Byambajav Buyandelger, Ching-Hsin Ku, Stefan Neef, Monika Bug, Katrin Schäfer, Gudrun Knöll, Leanne E. Felkin, Johannes Wessels, Karl Toischer, Franz Hagn, Horst Kessler, Michael Didié, Thomas Quentin, Lars S. Maier, Nils Teucher, Bernhard Unsöld, Albrecht Schmidt, Emma J. Birks, Sylvia Gunkel, Patrick Lang, Henk Granzier, Wolfram-Hubertus Zimmermann, Loren J. Field, Georgine Faulkner, Matthias Dobbelstein, Paul J.R. Barton, Michael Sattler, Matthias Wilmanns, Kenneth R. Chien
Circulation Research September 2011, 109 (7) 758-769; DOI: https://doi.org/10.1161/CIRCRESAHA.111.245787
By RalphKnöll, Wolfgang A.Linke, PeijianZou, SnježanaMiočiċ, SawaKostin, ByambajavBuyandelger, Ching-HsinKu, StefanNeef, MonikaBug, KatrinSchäfer, GudrunKnöll, Leanne E.Felkin, JohannesWessels, KarlToischer, FranzHagn, HorstKessler, MichaelDidié, ThomasQuentin, Lars S.Maier, NilsTeucher, BernhardUnsöld, AlbrechtSchmidt, Emma J.Birks, SylviaGunkel, PatrickLang, HenkGranzier, Wolfram-HubertusZimmermann, Loren J.Field, GeorgineFaulkner, MatthiasDobbelstein, Paul J.R.Barton, MichaelSattler, MatthiasWilmanns and Kenneth R.Chien
Generation of telethonin−/− animals. A, General strategy for gene targeting. The gene for telethonin is encoded by 2 exons; restriction sites are indi...Show More
Generation of telethonin−/− animals. A, General strategy for gene targeting. The gene for telethonin is encoded by 2 exons; restriction sites are indicated. The gene was replaced by a LacZ/Neomycin cassette (targeting construct is indicated). B, Southern hybridization of embryonic stem cells (left panel: different stem cell lines marked 1 to 6) as well as of resulting animals (right panel: different animals marked A–K). C, Telethonin mRNA expression, analyzed by Northern blot (upper row), as well as protein expression, analyzed by Western blotting, indicates that telethonin−/− results in a “true null allele.”Show Less
Ralph Knöll, Wolfgang A. Linke, Peijian Zou, Snježana Miočiċ, Sawa Kostin, Byambajav Buyandelger, Ching-Hsin Ku, Stefan Neef, Monika Bug, Katrin Schäfer, Gudrun Knöll, Leanne E. Felkin, Johannes Wessels, Karl Toischer, Franz Hagn, Horst Kessler, Michael Didié, Thomas Quentin, Lars S. Maier, Nils Teucher, Bernhard Unsöld, Albrecht Schmidt, Emma J. Birks, Sylvia Gunkel, Patrick Lang, Henk Granzier, Wolfram-Hubertus Zimmermann, Loren J. Field, Georgine Faulkner, Matthias Dobbelstein, Paul J.R. Barton, Michael Sattler, Matthias Wilmanns, Kenneth R. Chien
Circulation Research September 2011, 109 (7) 758-769; DOI: https://doi.org/10.1161/CIRCRESAHA.111.245787
By RalphKnöll, Wolfgang A.Linke, PeijianZou, SnježanaMiočiċ, SawaKostin, ByambajavBuyandelger, Ching-HsinKu, StefanNeef, MonikaBug, KatrinSchäfer, GudrunKnöll, Leanne E.Felkin, JohannesWessels, KarlToischer, FranzHagn, HorstKessler, MichaelDidié, ThomasQuentin, Lars S.Maier, NilsTeucher, BernhardUnsöld, AlbrechtSchmidt, Emma J.Birks, SylviaGunkel, PatrickLang, HenkGranzier, Wolfram-HubertusZimmermann, Loren J.Field, GeorgineFaulkner, MatthiasDobbelstein, Paul J.R.Barton, MichaelSattler, MatthiasWilmanns and Kenneth R.Chien
Probing functional consequences of telethonin deficiency at the subcellular and organismic levels. A and B, Myofibrils were isolated from either wildt...Show More
Probing functional consequences of telethonin deficiency at the subcellular and organismic levels. A and B, Myofibrils were isolated from either wildtype (WT) or homozygous telethonin-deficient (KO) mouse hearts and stretched to a desired sarcomere length (SL) under nonactivating conditions. Then, myofibrils were stained with an antibody to the telethonin-binding titin domains, Z1-Z2, and the secondary ones labeled using FITC-conjugated IgG. A, Phase-contrast (pc) and immunofluorescence (Z1Z2) images of stretched myofibrils from cardiac muscle before actin extraction; telethonin-deficient skeletal muscle; and cardiac muscle after actin extraction using a Ca2+-independent gelsolin fragment (shown are myofibrils at 2 different stretch states). Scale bar, 2 μm. B, (top) Quantitation of the broadness of the titin label in the Z-disk by determining the full width at half-maximum (FWHM) peak height on intensity profiles along the myofibril axis. (bottom) Average widths of Z1Z2–titin label in WT and KO cardiac myofibrils at different SLs, before and after actin extraction. Data are means ± SD (n=3 to 6). *P<0.05 in Student t test. C, Pull-down with MLP. N-terminus of titin (Z1Z2, used as a control), Tel (1 to 90), Tel (1 to 90, dE13), Tel (1 to 90, E13A), Tel (1 to 90, E13R), and Tel (1 to 90, E13W) were incubated with a recombinant GST-MLP fusion protein and pulled down with glutathione-sepharose 4B beads (anti-GST antibody antirabbit, Pharmacia Biotech, Sweden). D, Pull-down with Z1Z2. Same experiment as in C, except that instead of MLP an H-tagged N-terminus of titin Z1Z2 was used (pull down with Ni2+-NTA beads (QIAGEN, Germany), blot with antibody against telethonin). E, Native PAGE analysis of titin/telethonin complexes formed from telethonin and its mutants with Z1Z2. On the native gel, only the Z1Z2–telethonin complex and Z1Z2 were visible. F, Analysis of the Z1Z2–telethonin complex formation by size exclusion chromatography in combination with static light scattering. The complexes were loaded onto a sephadex column, molecular masses were calculated to be 23.0 (Z1Z2) and 55.4 (Z1Z2-telethonin complex) kDa. G, Structure of the telethonin–titin Z1-Z2 complex, the arrow indicates glutamate 13 (E13), important for stabilizing the β-hairpin structure. H, Functional analysis of telethonin deficiency in vivo: 2 to 3 weeks after transverse aortic constriction (TAC), telethonin−/− animals developed a defect in myocardial function (increased end-systolic and end-diastolic diameters, decrease in fractional shortening as well as increased left ventricular mass [LVM] and LVM per body mass [*P<0.05, **P<0.01], error bars indicate standard error of the mean [SEM]).Show Less
Ralph Knöll, Wolfgang A. Linke, Peijian Zou, Snježana Miočiċ, Sawa Kostin, Byambajav Buyandelger, Ching-Hsin Ku, Stefan Neef, Monika Bug, Katrin Schäfer, Gudrun Knöll, Leanne E. Felkin, Johannes Wessels, Karl Toischer, Franz Hagn, Horst Kessler, Michael Didié, Thomas Quentin, Lars S. Maier, Nils Teucher, Bernhard Unsöld, Albrecht Schmidt, Emma J. Birks, Sylvia Gunkel, Patrick Lang, Henk Granzier, Wolfram-Hubertus Zimmermann, Loren J. Field, Georgine Faulkner, Matthias Dobbelstein, Paul J.R. Barton, Michael Sattler, Matthias Wilmanns, Kenneth R. Chien
Circulation Research September 2011, 109 (7) 758-769; DOI: https://doi.org/10.1161/CIRCRESAHA.111.245787
By RalphKnöll, Wolfgang A.Linke, PeijianZou, SnježanaMiočiċ, SawaKostin, ByambajavBuyandelger, Ching-HsinKu, StefanNeef, MonikaBug, KatrinSchäfer, GudrunKnöll, Leanne E.Felkin, JohannesWessels, KarlToischer, FranzHagn, HorstKessler, MichaelDidié, ThomasQuentin, Lars S.Maier, NilsTeucher, BernhardUnsöld, AlbrechtSchmidt, Emma J.Birks, SylviaGunkel, PatrickLang, HenkGranzier, Wolfram-HubertusZimmermann, Loren J.Field, GeorgineFaulkner, MatthiasDobbelstein, Paul J.R.Barton, MichaelSattler, MatthiasWilmanns and Kenneth R.Chien
Telethonin—analysis of fibrosis, apoptosis, and p53. A, Wildtype (WT) and telethonin knockout (KO) hearts were analyzed for the presence of fibrosis (...Show More
Telethonin—analysis of fibrosis, apoptosis, and p53. A, Wildtype (WT) and telethonin knockout (KO) hearts were analyzed for the presence of fibrosis (masson trichrome stain) without intervention and after transverse aortic constriction (TAC). Telethonin transgenic animals did not develop any significant increase in fibrosis. B, Quantification of fibrosis; note the significant increase in fibrosis in the telethonin-deficient animals (without intervention [solid bars] and after TAC [open bars]; error bars indicate standard deviation [SD]). C, Wildtype (WT) and telethonin knockout (KO) hearts were analyzed for the presence of apoptosis without intervention and after transverse aortic constriction (TAC). D, Quantification of apoptosis; note the significant increase in apoptosis in the telethonin-deficient animals (without intervention [solid bars] and after TAC [open bars]; error bars indicate standard deviation [SD]). E, Western blot analysis of p53 expression in hearts of telethonin knockout as well as corresponding wildtype litter mate control hearts. Equal loading of the membrane has been confirmed by GAPDH gene expression. Note the significant increase in p53 expression in the telethonin−/− animals. F, Quantification of p53 protein expression. Data have been normalized to GAPDH. Note the significant increase in p53 expression in the telethonin−/− animals (n=4 animals per group; open bars: wildtype animals; solid bars: telethonin−/− animals. *P<0.05; error bars indicate standard deviation [SD]). G, Relative levels of p21 mRNA transcripts in left ventricles. We used hearts obtained from WT and telethonin−/− mice subjected to TAC for 3 weeks and analyzed mRNA expression by quantitative real-time PCR. Note the significant increase in p21 gene expression, which is a p53 target gene (open bars: wildtype animals [n=5]; solid bars: telethonin−/− animals [n=12]. *P<0.05 against WT-TAC; error bars indicate standard deviation [SD]).Show Less
