Systolic dysfunction in cardiac-specific ligand-inducible MerCreMer transgenic mice - PubMed (original) (raw)
Systolic dysfunction in cardiac-specific ligand-inducible MerCreMer transgenic mice
Michael E Hall et al. Am J Physiol Heart Circ Physiol. 2011 Jul.
Abstract
The Cre-loxP system is a useful tool to study the physiological effects of gene knockout in the heart. One limitation with using this system in the heart is the toxic effect of chronic expression of the Cre recombinase. To circumvent this limitation, a widely used inducible cardiac-specific model, Myh6-MerCreMer (Cre), using tamoxifen (TAM) to activate Cre has been developed. The current study examined cardiac function in Cre-positive C57B/J6 mice exposed to one, three, or five daily doses of a 40 mg/kg TAM to induce Cre activity specifically in the heart. Echocardiography demonstrated no statistically significant differences in systolic function (SF) at baseline as assessed by fractional shortening. In mice exposed to five injections, a significant fall in all determinants of SF was observed 6 days after TAM was initiated. However, SF returned to baseline levels 10 days after TAM initiation although the hearts exhibited significant hypertrophy. Heart weight-to-tibia length ratios were 73 ± 3, 78.5 ± 6, and 87.6 ± 9 mg/cm for one, three, and five TAM injections, respectively. TAM had no effect on cardiac function or hypertrophy in Cre-negative mice. Cre-positive mice receiving five TAM injections had significant reductions in cardiac mitochondrial ATP and significant reductions in the expression of proteins important for the regulation of cardiac oxidative phosphorylation including peroxisome proliferator-activated receptor-γ coactivator-1α and pyruvate dehydrogenase kinase-4. Thus inducible cardiac-specific activation of Cre recombinase caused a transient decline in SF that was dependent on the number of TAM doses and associated with significant hypertrophy and alterations in mitochondrial ATP and important proteins involved in the regulation of cardiac oxidative phosphorylation.
Figures
Fig. 1.
Systolic function as represented by fractional shortening following in 1-, 3-, or 5-day tamoxifen (TAM) treatment in animals expressing _Myh6_-MerCreMer (Cre+) or non-Cre (Cre−) mice. Day 0 is the average of the 2 baseline measurements and days 2, 4, 6, 8, and 10 represent days following the start of TAM administration. *P < 0.05 vs. Cre−/5-day mice.
Fig. 2.
Systolic and diastolic left ventricular dimensions for wall thickness and diameter obtained from M-mode measurements of echocardiography following in 1-, 3-, or 5-day tamoxifen treatment in animals expressing myosin heavy chain-MerCreMer or non-Cre controls. A: left ventricular anterior wall systole (LVAWs). B: left ventricular posterior wall systole (LVPWs). C: left ventricular anterior wall diastole (LVAWd). D: left ventricular posterior wall diastole (LVPWd). E: left ventricular end systolic diameter (LVESD). F: left ventricular end diastolic diameter (LVEDD). *P < 0.05 Cre+/5-day vs. Cre−/5-day mice. †P < 0.05 Cre+/3-day vs. Cre−/5-day mice.
Fig. 3.
Cardiac mitochondrial ATP levels determined in Cre−/5-day (n = 6) and Cre+/5-day (n = 8) mice at systolic dysfunction nadir (day 6) and at systolic function recovery (day 14). *P < 0.05, compared with Cre−/5-day mice.
Fig. 4.
Cardiac phosphorylated (p)-AMPK/total (t)AMPK protein levels in Cre−/5-day (n = 4) and Cre+/5-day (n = 4) mice measured by Western blot analysis. *P < 0.05, compared with Cre−/5-day, day 6. †P < 0.05, compared with Cre+/5-day, day 6. Bars represent breaks in original image separating individual groups that were run in quadruplicate.
Fig. 5.
Cardiac peroxisome proliferator-activated receptor-γ coactivator-1α (PGC1-α) protein levels in Cre−/5-day (n = 4) and Cre+/5-day (n = 4) mice measured by Western blot analysis. *P < 0.05, compared with Cre−/5-day mice. Bars represent breaks in original image separating individual groups that were run in quadruplicate.
Fig. 6.
Cardiac mitochondrial pyruvate dehydrogenase kinase-4 (PDK-4) protein levels in Cre−/5-day (n = 4) and Cre+/5-day (n = 4) mice measured by Western blot analysis. *P < 0.05, compared with Cre−/5-day, day 6. Bars represent breaks in original image separating individual groups that were run in quadruplicate.
Fig. 7.
Cardiac β-tubulin protein levels in Cre−/5-day (n = 4) and Cre+/5-day (n = 4) mice measured by Western blot analysis. P < 0.05, compared with Cre−/5-day, day 6. Bars represent breaks in original image separating individual groups that were run in quadruplicate.
Fig. 8.
Cardiac histology from Cre+/5-day and Cre−/5-day mice at day 6 and day 10 post-TAM treatment. A, D, and G: hemotoxylin and eosin (H&E) staining at ×200 in untreated Cre− (A); Cre+/5-day, day 6 (D); and Cre+/5-day, day 10 (G). B, E, and H: periodic acid-Schiff (PAS) staining at ×200 in untreated Cre− (B); Cre+/5-day, day 6 (E); and Cre+/5-day, day 10 (H). C, F, and I: picrosirius red (PSR) staining at ×200 in untreated Cre− (C); Cre+/5-day, day 6 (F); and Cre+/5-day, day 10 (I). H&E staining reveals inflammatory cells seen on day 6 (‡) and condensed nuclear chromatin noted at Systolic function recovery (*). At day 10, intracellular staining on PAS (#) and appearance of increase collagen on PSR (†).
Similar articles
- _Myh6_-driven Cre recombinase activates the DNA damage response and the cell cycle in the myocardium in the absence of loxP sites.
Wang X, Lauth A, Wan TC, Lough JW, Auchampach JA. Wang X, et al. Dis Model Mech. 2020 Dec 18;13(12):dmm046375. doi: 10.1242/dmm.046375. Dis Model Mech. 2020. PMID: 33106234 Free PMC article. - 4-hydroxytamoxifen does not deteriorate cardiac function in cardiomyocyte-specific MerCreMer transgenic mice.
Heinen A, Gödecke S, Flögel U, Miklos D, Bottermann K, Spychala A, Gödecke A. Heinen A, et al. Basic Res Cardiol. 2021 Feb 5;116(1):8. doi: 10.1007/s00395-020-00841-9. Basic Res Cardiol. 2021. PMID: 33544211 Free PMC article. - Cardiomyocyte-specific deletion of leptin receptors causes lethal heart failure in Cre-recombinase-mediated cardiotoxicity.
Hall ME, Smith G, Hall JE, Stec DE. Hall ME, et al. Am J Physiol Regul Integr Comp Physiol. 2012 Dec 15;303(12):R1241-50. doi: 10.1152/ajpregu.00292.2012. Epub 2012 Oct 31. Am J Physiol Regul Integr Comp Physiol. 2012. PMID: 23115124 Free PMC article. - Temporally regulated and tissue-specific gene manipulations in the adult and embryonic heart using a tamoxifen-inducible Cre protein.
Sohal DS, Nghiem M, Crackower MA, Witt SA, Kimball TR, Tymitz KM, Penninger JM, Molkentin JD. Sohal DS, et al. Circ Res. 2001 Jul 6;89(1):20-5. doi: 10.1161/hh1301.092687. Circ Res. 2001. PMID: 11440973 - A review of tamoxifen administration regimen optimization for Cre/loxp system in mouse bone study.
Chen MY, Zhao FL, Chu WL, Bai MR, Zhang DM. Chen MY, et al. Biomed Pharmacother. 2023 Sep;165:115045. doi: 10.1016/j.biopha.2023.115045. Epub 2023 Jun 28. Biomed Pharmacother. 2023. PMID: 37379643 Review.
Cited by
- α-Melanocyte-stimulating hormone alleviates pathological cardiac remodeling via melanocortin 5 receptor.
Suominen A, Saldo Rubio G, Ruohonen S, Szabó Z, Pohjolainen L, Ghimire B, Ruohonen ST, Saukkonen K, Ijas J, Skarp S, Kaikkonen L, Cai M, Wardlaw SL, Ruskoaho H, Talman V, Savontaus E, Kerkelä R, Rinne P. Suominen A, et al. EMBO Rep. 2024 Apr;25(4):1987-2014. doi: 10.1038/s44319-024-00109-6. Epub 2024 Mar 7. EMBO Rep. 2024. PMID: 38454158 Free PMC article. - Myosin Light Chain Phosphatase Plays an Important Role in Cardiac Fibrosis in a Model of Mineralocorticoid Receptor-Associated Hypertension.
Ye Z, Okamoto R, Ito H, Ito R, Moriwaki K, Ichikawa M, Kimena L, Ali Y, Ito M, Gomez-Sanchez CE, Dohi K. Ye Z, et al. J Am Heart Assoc. 2024 Mar 5;13(5):e032828. doi: 10.1161/JAHA.123.032828. Epub 2024 Feb 29. J Am Heart Assoc. 2024. PMID: 38420846 Free PMC article. - Prolonged tamoxifen-enriched diet is associated with cardiomyopathy and nutritional frailty in mice.
Halpage J, DaSilva Pantoja P, Mancarella S. Halpage J, et al. Exp Physiol. 2024 Apr;109(4):513-523. doi: 10.1113/EP091668. Epub 2024 Jan 30. Exp Physiol. 2024. PMID: 38291801 Free PMC article. - Perspective: The current state of Cre driver mouse lines in skeletal research: Challenges and opportunities.
Cunningham CJ, Choi RB, Bullock WA, Robling AG. Cunningham CJ, et al. Bone. 2023 May;170:116719. doi: 10.1016/j.bone.2023.116719. Epub 2023 Mar 1. Bone. 2023. PMID: 36868507 Free PMC article. Review. - Cardiac-Specific Expression of Cre Recombinase Leads to Age-Related Cardiac Dysfunction Associated with Tumor-like Growth of Atrial Cardiomyocyte and Ventricular Fibrosis and Ferroptosis.
Li Z, Duan Q, Cui Y, Jones OD, Shao D, Zhang J, Gao Y, Cao X, Wang S, Li J, Lei X, Zhang W, Wang L, Zhou X, Xu M, Liu Y, Ma J, Xu X. Li Z, et al. Int J Mol Sci. 2023 Feb 4;24(4):3094. doi: 10.3390/ijms24043094. Int J Mol Sci. 2023. PMID: 36834504 Free PMC article.
References
- Andersson KB, Birkeland JA, Finsen AV, Louch WE, Sjaastad I, Wang Y, Chen J, Molkentin JD, Chien KR, Sejersted OM, Christensen G. Moderate heart dysfunction in mice with inducible cardiomyocyte-specific excision of the Serca2 gene. J Mol Cell Cardiol 47: 180–187, 2009 - PubMed
- Andersson KB, Winer LH, Mork HK, Molkentin JD, Jaisser F. Tamoxifen administration routes and dosage for inducible Cre-mediated gene disruption in mouse hearts. Transgenic Res 19: 715–725, 2010 - PubMed
- Buerger A, Rozhitskaya O, Sherwood MC, Dorfman AL, Bisping E, Abel ED, Pu WT, Izumo S, Jay PY. Dilated cardiomyopathy resulting from high-level myocardial expression of Cre-recombinase. J Card Fail 12: 392–398, 2006 - PubMed
- Costa Martins PA, Bourajjaj M, Gladka M, Kortland M, van Oort RJ, Pinto YM, Molkentin JD, De Windt LJ. Conditional dicer gene deletion in the postnatal myocardium provokes spontaneous cardiac remodeling. Circulation 118: 1567–1576, 2008 - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Molecular Biology Databases