Creatine uptake in mouse hearts with genetically altered creatine levels
Creatine plays an important role in energy metabolism in the heart. Cardiomyocytes accumulate creatine via a specific creatine transporter (CrT), the capacity of which is reduced in the failing heart, resulting in lower myocardial creatine concentration. Therefore, to gain insight into how the CrT i...
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| Published in | Journal of molecular and cellular cardiology Vol. 45; no. 3; pp. 453 - 459 |
|---|---|
| Main Authors | , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
England
Elsevier Ltd
01.09.2008
Academic Press |
| Subjects | |
| Online Access | Get full text |
| ISSN | 0022-2828 1095-8584 1095-8584 |
| DOI | 10.1016/j.yjmcc.2008.05.023 |
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| Abstract | Creatine plays an important role in energy metabolism in the heart. Cardiomyocytes accumulate creatine via a specific creatine transporter (CrT), the capacity of which is reduced in the failing heart, resulting in lower myocardial creatine concentration. Therefore, to gain insight into how the CrT is regulated, we studied two mouse models of severely altered myocardial creatine levels. Cardiac creatine uptake levels were measured in isolated hearts from creatine-free guanidinoacetate-N-methyl transferase knock out (GAMT−/−) mice and from mice overexpressing the myocardial CrT (CrT-OE) using 14C-radiolabeled creatine. CrT mRNA levels were measured using real time RT-PCR and creatine levels with HPLC. Hearts from GAMT−/− mice showed a 7-fold increase in Vmax of creatine uptake and a 1.4-fold increase in CrT mRNA levels. The increase in Cr uptake and in CrT mRNA levels, however, was almost completely prevented when mice were fed a creatine supplemented diet, indicating that creatine uptake is subject to negative feedback regulation. Cardiac creatine uptake levels in CrT-OE mice were increased on average 2.7-fold, showing a considerable variation, in line with a similar variation in creatine content. Total CrT mRNA levels correlated well with myocardial creatine content (r=0.67; p<0.0001) but endogenous CrT mRNA levels did not correlate at all with myocardial creatine content (r=0.01; p=0.96). This study shows that creatine uptake can be massively upregulated in the heart, by almost an order of magnitude and that this upregulation is subject to feedback inhibition. In addition, our results strongly suggest that CrT activity is predominantly regulated by mechanisms other than alterations in gene expression. |
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| AbstractList | Creatine plays an important role in energy metabolism in the heart. Cardiomyocytes accumulate creatine via a specific creatine transporter (CrT), the capacity of which is reduced in the failing heart, resulting in lower myocardial creatine concentration. Therefore, to gain insight into how the CrT is regulated, we studied two mouse models of severely altered myocardial creatine levels. Cardiac creatine uptake levels were measured in isolated hearts from creatine-free guanidinoacetate-N-methyl transferase knock out (GAMT(-/-)) mice and from mice overexpressing the myocardial CrT (CrT-OE) using (14)C-radiolabeled creatine. CrT mRNA levels were measured using real time RT-PCR and creatine levels with HPLC. Hearts from GAMT(-/-) mice showed a 7-fold increase in V(max) of creatine uptake and a 1.4-fold increase in CrT mRNA levels. The increase in Cr uptake and in CrT mRNA levels, however, was almost completely prevented when mice were fed a creatine supplemented diet, indicating that creatine uptake is subject to negative feedback regulation. Cardiac creatine uptake levels in CrT-OE mice were increased on average 2.7-fold, showing a considerable variation, in line with a similar variation in creatine content. Total CrT mRNA levels correlated well with myocardial creatine content (r=0.67; p<0.0001) but endogenous CrT mRNA levels did not correlate at all with myocardial creatine content (r=0.01; p=0.96). This study shows that creatine uptake can be massively upregulated in the heart, by almost an order of magnitude and that this upregulation is subject to feedback inhibition. In addition, our results strongly suggest that CrT activity is predominantly regulated by mechanisms other than alterations in gene expression.Creatine plays an important role in energy metabolism in the heart. Cardiomyocytes accumulate creatine via a specific creatine transporter (CrT), the capacity of which is reduced in the failing heart, resulting in lower myocardial creatine concentration. Therefore, to gain insight into how the CrT is regulated, we studied two mouse models of severely altered myocardial creatine levels. Cardiac creatine uptake levels were measured in isolated hearts from creatine-free guanidinoacetate-N-methyl transferase knock out (GAMT(-/-)) mice and from mice overexpressing the myocardial CrT (CrT-OE) using (14)C-radiolabeled creatine. CrT mRNA levels were measured using real time RT-PCR and creatine levels with HPLC. Hearts from GAMT(-/-) mice showed a 7-fold increase in V(max) of creatine uptake and a 1.4-fold increase in CrT mRNA levels. The increase in Cr uptake and in CrT mRNA levels, however, was almost completely prevented when mice were fed a creatine supplemented diet, indicating that creatine uptake is subject to negative feedback regulation. Cardiac creatine uptake levels in CrT-OE mice were increased on average 2.7-fold, showing a considerable variation, in line with a similar variation in creatine content. Total CrT mRNA levels correlated well with myocardial creatine content (r=0.67; p<0.0001) but endogenous CrT mRNA levels did not correlate at all with myocardial creatine content (r=0.01; p=0.96). This study shows that creatine uptake can be massively upregulated in the heart, by almost an order of magnitude and that this upregulation is subject to feedback inhibition. In addition, our results strongly suggest that CrT activity is predominantly regulated by mechanisms other than alterations in gene expression. Creatine plays an important role in energy metabolism in the heart. Cardiomyocytes accumulate creatine via a specific creatine transporter (CrT), the capacity of which is reduced in the failing heart, resulting in lower myocardial creatine concentration. Therefore, to gain insight into how the CrT is regulated, we studied two mouse models of severely altered myocardial creatine levels. Cardiac creatine uptake levels were measured in isolated hearts from creatine-free guanidinoacetate-N-methyl transferase knock out (GAMT−/−) mice and from mice overexpressing the myocardial CrT (CrT-OE) using 14C-radiolabeled creatine. CrT mRNA levels were measured using real time RT-PCR and creatine levels with HPLC. Hearts from GAMT−/− mice showed a 7-fold increase in Vmax of creatine uptake and a 1.4-fold increase in CrT mRNA levels. The increase in Cr uptake and in CrT mRNA levels, however, was almost completely prevented when mice were fed a creatine supplemented diet, indicating that creatine uptake is subject to negative feedback regulation. Cardiac creatine uptake levels in CrT-OE mice were increased on average 2.7-fold, showing a considerable variation, in line with a similar variation in creatine content. Total CrT mRNA levels correlated well with myocardial creatine content (r=0.67; p<0.0001) but endogenous CrT mRNA levels did not correlate at all with myocardial creatine content (r=0.01; p=0.96). This study shows that creatine uptake can be massively upregulated in the heart, by almost an order of magnitude and that this upregulation is subject to feedback inhibition. In addition, our results strongly suggest that CrT activity is predominantly regulated by mechanisms other than alterations in gene expression. Creatine plays an important role in energy metabolism in the heart. Cardiomyocytes accumulate creatine via a specific creatine transporter (CrT), the capacity of which is reduced in the failing heart, resulting in lower myocardial creatine concentration. Therefore, to gain insight into how the CrT is regulated, we studied two mouse models of severely altered myocardial creatine levels. Cardiac creatine uptake levels were measured in isolated hearts from creatine-free guanidinoacetate- N -methyl transferase knock out (GAMT −/− ) mice and from mice overexpressing the myocardial CrT (CrT-OE) using 14 C-radiolabeled creatine. CrT mRNA levels were measured using real time RT-PCR and creatine levels with HPLC. Hearts from GAMT −/− mice showed a 7-fold increase in V max of creatine uptake and a 1.4-fold increase in CrT mRNA levels. The increase in Cr uptake and in CrT mRNA levels, however, was almost completely prevented when mice were fed a creatine supplemented diet, indicating that creatine uptake is subject to negative feedback regulation. Cardiac creatine uptake levels in CrT-OE mice were increased on average 2.7-fold, showing a considerable variation, in line with a similar variation in creatine content. Total CrT mRNA levels correlated well with myocardial creatine content ( r = 0.67; p < 0.0001) but endogenous CrT mRNA levels did not correlate at all with myocardial creatine content ( r = 0.01; p = 0.96). This study shows that creatine uptake can be massively upregulated in the heart, by almost an order of magnitude and that this upregulation is subject to feedback inhibition. In addition, our results strongly suggest that CrT activity is predominantly regulated by mechanisms other than alterations in gene expression. Abstract Creatine plays an important role in energy metabolism in the heart. Cardiomyocytes accumulate creatine via a specific creatine transporter (CrT), the capacity of which is reduced in the failing heart, resulting in lower myocardial creatine concentration. Therefore, to gain insight into how the CrT is regulated, we studied two mouse models of severely altered myocardial creatine levels. Cardiac creatine uptake levels were measured in isolated hearts from creatine-free guanidinoacetate- N -methyl transferase knock out (GAMT−/− ) mice and from mice overexpressing the myocardial CrT (CrT-OE) using14 C-radiolabeled creatine. CrT mRNA levels were measured using real time RT-PCR and creatine levels with HPLC. Hearts from GAMT−/− mice showed a 7-fold increase in Vmax of creatine uptake and a 1.4-fold increase in CrT mRNA levels. The increase in Cr uptake and in CrT mRNA levels, however, was almost completely prevented when mice were fed a creatine supplemented diet, indicating that creatine uptake is subject to negative feedback regulation. Cardiac creatine uptake levels in CrT-OE mice were increased on average 2.7-fold, showing a considerable variation, in line with a similar variation in creatine content. Total CrT mRNA levels correlated well with myocardial creatine content ( r = 0.67; p < 0.0001) but endogenous CrT mRNA levels did not correlate at all with myocardial creatine content ( r = 0.01; p = 0.96). This study shows that creatine uptake can be massively upregulated in the heart, by almost an order of magnitude and that this upregulation is subject to feedback inhibition. In addition, our results strongly suggest that CrT activity is predominantly regulated by mechanisms other than alterations in gene expression. Creatine plays an important role in energy metabolism in the heart. Cardiomyocytes accumulate creatine via a specific creatine transporter (CrT), the capacity of which is reduced in the failing heart, resulting in lower myocardial creatine concentration. Therefore, to gain insight into how the CrT is regulated, we studied two mouse models of severely altered myocardial creatine levels. Cardiac creatine uptake levels were measured in isolated hearts from creatine-free guanidinoacetate-N-methyl transferase knock out (GAMT(-/-)) mice and from mice overexpressing the myocardial CrT (CrT-OE) using (14)C-radiolabeled creatine. CrT mRNA levels were measured using real time RT-PCR and creatine levels with HPLC. Hearts from GAMT(-/-) mice showed a 7-fold increase in V(max) of creatine uptake and a 1.4-fold increase in CrT mRNA levels. The increase in Cr uptake and in CrT mRNA levels, however, was almost completely prevented when mice were fed a creatine supplemented diet, indicating that creatine uptake is subject to negative feedback regulation. Cardiac creatine uptake levels in CrT-OE mice were increased on average 2.7-fold, showing a considerable variation, in line with a similar variation in creatine content. Total CrT mRNA levels correlated well with myocardial creatine content (r=0.67; p<0.0001) but endogenous CrT mRNA levels did not correlate at all with myocardial creatine content (r=0.01; p=0.96). This study shows that creatine uptake can be massively upregulated in the heart, by almost an order of magnitude and that this upregulation is subject to feedback inhibition. In addition, our results strongly suggest that CrT activity is predominantly regulated by mechanisms other than alterations in gene expression. |
| Author | Hunyor, Imre Lygate, Craig Hove, Michiel ten Isbrandt, Dirk Wallis, Julie Neubauer, Stefan Sebag-Montefiore, Liam Fischer, Alexandra Makinen, Kimmo |
| AuthorAffiliation | b Center for Molecular Neurobiology, University of Hamburg, Hamburg, Germany a Department of Cardiovascular Medicine, University of Oxford, Oxford, UK |
| AuthorAffiliation_xml | – name: a Department of Cardiovascular Medicine, University of Oxford, Oxford, UK – name: b Center for Molecular Neurobiology, University of Hamburg, Hamburg, Germany |
| Author_xml | – sequence: 1 givenname: Michiel ten surname: Hove fullname: Hove, Michiel ten email: michiel.tenhove@gmail.com, michiel.tenhove@well.ox.ac.uk organization: Department of Cardiovascular Medicine, University of Oxford, Oxford, UK – sequence: 2 givenname: Kimmo surname: Makinen fullname: Makinen, Kimmo organization: Department of Cardiovascular Medicine, University of Oxford, Oxford, UK – sequence: 3 givenname: Liam surname: Sebag-Montefiore fullname: Sebag-Montefiore, Liam organization: Department of Cardiovascular Medicine, University of Oxford, Oxford, UK – sequence: 4 givenname: Imre surname: Hunyor fullname: Hunyor, Imre organization: Department of Cardiovascular Medicine, University of Oxford, Oxford, UK – sequence: 5 givenname: Alexandra surname: Fischer fullname: Fischer, Alexandra organization: Department of Cardiovascular Medicine, University of Oxford, Oxford, UK – sequence: 6 givenname: Julie surname: Wallis fullname: Wallis, Julie organization: Department of Cardiovascular Medicine, University of Oxford, Oxford, UK – sequence: 7 givenname: Dirk surname: Isbrandt fullname: Isbrandt, Dirk organization: Center for Molecular Neurobiology, University of Hamburg, Hamburg, Germany – sequence: 8 givenname: Craig surname: Lygate fullname: Lygate, Craig organization: Department of Cardiovascular Medicine, University of Oxford, Oxford, UK – sequence: 9 givenname: Stefan surname: Neubauer fullname: Neubauer, Stefan organization: Department of Cardiovascular Medicine, University of Oxford, Oxford, UK |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/18602925$$D View this record in MEDLINE/PubMed |
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| Cites_doi | 10.1093/nar/gng154 10.1161/01.CIR.0000165147.99592.01 10.1161/CIRCULATIONAHA.105.572990 10.1152/physrev.2000.80.3.1107 10.1016/0005-2728(96)00018-7 10.1006/jmcc.1996.0143 10.1161/01.RES.0000137170.41939.d9 10.1016/j.yjmcc.2004.11.016 10.1073/pnas.85.3.807 10.1006/geno.1996.0373 10.1093/hmg/ddh112 10.1074/jbc.274.50.35794 10.1056/NEJMra063052 10.1016/j.bbrc.2005.06.164 10.1172/JCI117756 10.1016/S0021-9258(19)49616-6 10.1016/j.yjmcc.2007.03.899 10.1023/B:MCBI.0000009886.98508.e7 |
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| Keywords | Creatine transport GAMT Energy metabolism Creatine Transgenic mouse model |
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| Snippet | Creatine plays an important role in energy metabolism in the heart. Cardiomyocytes accumulate creatine via a specific creatine transporter (CrT), the capacity... Abstract Creatine plays an important role in energy metabolism in the heart. Cardiomyocytes accumulate creatine via a specific creatine transporter (CrT), the... |
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| SubjectTerms | Animals Biological Transport - physiology Cardiovascular Creatine Creatine - genetics Creatine - metabolism Creatine transport Energy metabolism Female GAMT Guanidinoacetate N-Methyltransferase - deficiency Guanidinoacetate N-Methyltransferase - genetics Humans Male Mice Mice, Inbred C57BL Mice, Knockout Mice, Transgenic Myocardium - metabolism Original Rabbits Transgenic mouse model |
| Title | Creatine uptake in mouse hearts with genetically altered creatine levels |
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