Impaired muscular contractile performance and adenine nucleotide handling in creatine kinase-deficient mice

2001 ◽  
Vol 281 (3) ◽  
pp. E619-E625 ◽  
Author(s):  
M. Gorselink ◽  
M. R. Drost ◽  
W. A. Coumans ◽  
G. P. J. van Kranenburg ◽  
R. P. Hesselink ◽  
...  
Keyword(s):  
Creatine Kinase ◽  
Adenylate Kinase ◽  
Adenine Nucleotide ◽  
Recovery Period ◽  
High Energy ◽  
Wild Type ◽  
The Impact ◽  
Kinase Pathway ◽  
Contractile Performance ◽  
Atp Regeneration

Creatine kinase (CK) forms a small family of isoenzymes playing an important role in maintaining the concentration of ATP and ADP in muscle cells. To delineate the impact of a lack of CK activity, we studied contractile performance during a single maximal tetanic contraction and during 12 repeated tetanic contractions of intact dorsal flexors of CK knockout (CK−/−) mice. To investigate the effect on ATP regeneration, muscular high-energy phosphate content was determined at rest, immediately after the contraction series, and after a 60-s recovery period. Maximal torque of the dorsal flexors was significantly lower in CK−/− mice than in wild-type animals, i.e., 23.7 ± 5.1 and 33.3 ± 6.8 mN · m · g−1 wet wt, respectively. Lower muscle ATP (20.1 ± 1.4 in CK−/− vs. 28.0 ± 2.1 μmol/g dry wt in controls) and higher IMP (1.2 ± 0.5 in CK−/− vs. 0.3 ± 0.1 μmol/g dry wt in controls) levels at the onset of contraction may contribute to the declined contractility in CK−/− mice. In contrast to wild-type muscles, ATP levels could not be maintained during the series of 12 tetanic contractions of dorsal flexors of CK−/− mice and dropped to 15.5 ± 2.4 μmol/g dry wt. The significant increase in tissue IMP (2.4 ± 1.1 μmol/g dry wt) content after the contraction series indicates that ATP regeneration through adenylate kinase was not capable of fully compensating for the lack of CK. ATP regeneration via the adenylate kinase pathway is a likely cause of reduced basal adenine nucleotide levels in CK−/− mice.

Skeletal muscle contractile performance and ADP accumulation in adenylate kinase-deficient mice

2005 ◽  
Vol 288 (6) ◽  
pp. C1287-C1297 ◽  
Author(s):  
Chad R. Hancock ◽  
Edwin Janssen ◽  
Ronald L. Terjung
Keyword(s):  
Skeletal Muscle ◽  
Adenylate Kinase ◽  
Energy Demand ◽  
Adenine Nucleotide ◽  
Formation Rate ◽  
High Energy ◽  
Energy Demands ◽  
Whole Muscle ◽  
Muscle Contractile ◽  
Contractile Performance

The production of AMP by adenylate kinase (AK) and subsequent deamination by AMP deaminase limits ADP accumulation during conditions of high-energy demand in skeletal muscle. The goal of this study was to investigate the consequences of AK deficiency (−/−) on adenine nucleotide management and whole muscle function at high-energy demands. To do this, we examined isometric tetanic contractile performance of the gastrocnemius-plantaris-soleus (GPS) muscle group in situ in AK1−/− mice and wild-type (WT) controls over a range of contraction frequencies (30–120 tetani/min). We found that AK1−/− muscle exhibited a diminished inosine 5′-monophosphate formation rate (14% of WT) and an inordinate accumulation of ADP (∼1.5 mM) at the highest energy demands, compared with WT controls. AK-deficient muscle exhibited similar initial contractile performance (521 ± 9 and 521 ± 10 g tension in WT and AK1−/− muscle, respectively), followed by a significant slowing of relaxation kinetics at the highest energy demands relative to WT controls. This is consistent with a depressed capacity to sequester calcium in the presence of high ADP. However, the overall pattern of fatigue in AK1−/− mice was similar to WT control muscle. Our findings directly demonstrate the importance of AMP formation and subsequent deamination in limiting ADP accumulation. Whole muscle contractile performance was, however, remarkably tolerant of ADP accumulation markedly in excess of what normally occurs in skeletal muscle.

Diabetes decreases creatine kinase enzyme activity and mRNA level in the rat heart

1989 ◽  
Vol 257 (4) ◽  
pp. E573-E577 ◽  
Author(s):  
B. K. Popovich ◽  
K. R. Boheler ◽  
W. H. Dillmann
Keyword(s):  
Creatine Kinase ◽  
Enzyme Activity ◽  
Northern Blot Analysis ◽  
Mrna Level ◽  
Cdna Probe ◽  
High Energy ◽  
Diabetic Rats ◽  
Mrna Levels ◽  
High Energy Phosphates ◽  
Contractile Performance

Several of the adenosinetriphosphatase enzymes that are responsible for cardiac muscle contraction rely on high-energy phosphates supplied by the creatine kinase (CK) system. Experimental diabetes mellitus has been shown to cause a decrease in the maximal contractile performance of the heart. We postulated that the decrease in contractile performance may be explained in part by a decrease in CK enzyme activity. To evaluate this possibility, we determined the level of CK activity and isoenzyme distribution in ventricular homogenates from normal, diabetic, and insulin-treated diabetic rats. We found that total CK activity was decreased by 35% in diabetic hearts and that a 66% reduction in the cardiac-specific MB isoenzyme occurs. Using a cDNA probe for CK-muscle (M) RNA in Northern blot analysis, we determined that a 61.1% decrease in CK-M mRNA occurs in diabetes. Chronic insulin therapy for 1 mo restores CK-M mRNA levels and enzyme activity. In conclusion, diabetes-induced CK enzyme decreases are mediated in part by a lower level of CK-M mRNA that codes for the major CK-M subunit protein. Decreased performance of the CK system may contribute to diabetic cardiomyopathy.

Activation time of myocardial oxidative phosphorylation in creatine kinase and adenylate kinase knockout mice

2002 ◽  
Vol 282 (6) ◽  
pp. H2259-H2264 ◽  
Author(s):  
Lori A. Gustafson ◽  
Johannes H. G. M. Van Beek
Keyword(s):  
Heart Rate ◽  
Creatine Kinase ◽  
Oxygen Consumption ◽  
Response Time ◽  
Adenylate Kinase ◽  
Contractile Function ◽  
Left Ventricular ◽  
Wild Type ◽  
Dynamic Regulation ◽  
Developed Pressure

Our goal was to determine whether mice genetically altered to lack either creatine kinase (M/MtCK−/−) or adenylate kinase (AK−/−) show altered properties in the dynamic regulation of myocardial oxygen consumption (MV˙o 2). We measured contractile function, oxygen consumption, and the mean response time of oxygen consumption to a step increase in heart rate [i.e., mitochondrial response time ( t mito)] in isolated Langendorff-perfused hearts from wild-type ( n = 6), M/MtCK−/− ( n = 6), and AK−/− ( n = 4) mice. Left ventricular developed pressure was higher in M/MtCK−/− hearts (88.2 ± 6.8 mmHg) and lower in AK−/− hearts (46.7 ± 9.4 mmHg) compared with wild-type hearts (60.7 ± 10.1 mmHg) at the basal pacing rate. Developed pressure fell slightly when heart rate was increased in all three groups. Basal MV˙o 2 at 300 beats/min was 19.1 ± 2.4, 19.4 ± 1.5, and 16.3 ± 1.9 μmol · min−1 · g dry wt−1for M/MtCK−/−, AK−/−, and wild type, respectively, which increased to 25.5 ± 3.7, 25.4 ± 2.6, and 22.0 ± 2.6 μmol · min−1 · g−1, when heart rate was increased to 400 beats/min. The t mito was significantly faster in M/MtCK−/− hearts: 3.0 ± 0.3 versus 7.3 ± 0.6 and 8.0 ± 0.4 s for M/MtCK−/−, AK−/−, and wild-type hearts, respectively. Our results demonstrate that MV˙o 2 of M/MtCK−/− hearts adapts more quickly to an increase in heart rate and thereby support the hypothesis that creatine kinase acts as an energy buffer in the cytosol, which delays the energy-related signal between sites of ATP hydrolysis and mitochondria.

scholarly journals Mitochondrial creatine kinase isoform expression does not correlate with its mode of action

1997 ◽  
Vol 322 (1) ◽  
pp. 73-78 ◽  
Author(s):  
Keltoum ANFLOUS ◽  
Vladimir VEKSLER ◽  
Philippe MATEO ◽  
Françoise SAMSON ◽  
Valdour SAKS ◽  
...  
Keyword(s):  
Creatine Kinase ◽  
Smooth Muscle ◽  
Adenylate Kinase ◽  
Adenine Nucleotide ◽  
Electrophoretic Analysis ◽  
Intermembrane Space ◽  
Mitochondrial Creatine Kinase ◽  
Cardiac Tissues ◽  
Atp Production ◽  
Mitochondrial Inner Membrane

In adult mammalian ventricular tissue, mitochondrial creatine kinase (mi-CK), which is bound to the outer surface of the mitochondrial inner membrane, is functionally coupled to oxidative phosphorylation. This is shown, in saponin-permeabilized rat ventricular fibres, by a shift in the apparent Km of mitochondrial respiration for ADP from 300ŷ56 ƁM to 111ŷ40 ƁM (P < 0.001) on the addition of 25 mM creatine, due to a local accumulation of ADP close to the ATP/ADP translocator. We have found that, in atrial fibres, the apparent Km for ADP is high, but is not decreased by creatine, suggesting an absence of coupling in this tissue, as has previously been observed in smooth muscle. mi-CK is encoded by two different genes, which are expressed in a tissue-specific manner: the sarcomeric isoform is expressed in ventricular and skeletal muscles, while the ubiquitous isoform is expressed in smooth muscle, brain and other tissues. In order to determine whether a specific function can be attributed to the expression of a specific isoform, we investigated mi-CK mRNA expression by Northern blot analysis. Hybridization with synthetic oligonucleotides specific for each mi-CK isoform showed the expression of only the sarcomeric isoform in rat atria. This result was confirmed by PCR using primers specific for each isoform. In addition, electrophoretic analysis of CK isoforms showed no difference in the octamer/dimer ratio of mi-CK in the atria and ventricles. In atria, unlike the soleus or ventricles, the maximum potential rate of mitochondrial phosphocreatine synthesis was lower than the maximal rate of ATP production by the mitochondria. The total CK/adenylate kinase ratio was also lower in atria than in the other tissues, suggesting a greater contribution of adenylate kinase to adenine nucleotide compartmentation in this tissue. The functional differences between mi-CK in the two cardiac tissues seem to imply a specific arrangement of the proteins in the intermembrane space rather than the expression of specific isoforms.

Mitochondrial creatine kinase is critically necessary for normal myocardial high-energy phosphate metabolism

2002 ◽  
Vol 283 (2) ◽  
pp. H680-H687 ◽  
Author(s):  
Matthias Spindler ◽  
Reinhard Niebler ◽  
Helga Remkes ◽  
Michael Horn ◽  
Titus Lanz ◽  
...  
Keyword(s):  
Creatine Kinase ◽  
Energy Homeostasis ◽  
Atp Hydrolysis ◽  
High Energy ◽  
Left Ventricular ◽  
Critical Threshold ◽  
Wild Type ◽  
High Energy Phosphate ◽  
The Individual ◽  
High Energy Phosphate Metabolism

The individual functional significance of the various creatine kinase (CK) isoenzymes for myocardial energy homeostasis is poorly understood. Whereas transgenic hearts lacking the M subunit of CK (M-CK) show unaltered cardiac energetics and left ventricular (LV) performance, deletion of M-CK in combination with loss of sarcomeric mitochondrial CK (ScCKmit) leads to significant alterations in myocardial high-energy phosphate metabolites. To address the question as to whether this alteration is due to a decrease in total CK activity below a critical threshold or due to the specific loss of ScCKmit, we studied isolated perfused hearts with selective loss of ScCKmit (ScCKmit−/−, remaining total CK activity ∼70%) using31P NMR spectroscopy at two different workloads. LV performance in ScCKmit−/− hearts ( n = 11) was similar compared with wild-type hearts ( n = 9). Phosphocreatine/ATP, however, was significantly reduced in ScCKmit−/− compared with wild-type hearts (1.02 ± 0.05 vs. 1.54 ± 0.07, P < 0.05). In parallel, free [ADP] was higher (144 ± 11 vs. 67 ± 7 μM, P < 0.01) and free energy release for ATP hydrolysis (Δ G ATP) was lower (−55.8 ± 0.5 vs. −58.5 ± 0.5 kJ/mol, P < 0.01) in ScCKmit−/− compared with wild-type hearts. These results demonstrate that M- and B-CK containing isoenzymes are unable to fully substitute for the loss of ScCKmit. We conclude that ScCKmit, in contrast to M-CK, is critically necessary to maintain normal high-energy phosphate metabolite levels in the heart.

Mouse p53 represses the rat brain creatine kinase gene but activates the rat muscle creatine kinase gene

1994 ◽  
Vol 14 (12) ◽  
pp. 8483-8492
Author(s):  
J Zhao ◽  
F I Schmieg ◽  
D T Simmons ◽  
G R Molloy
Keyword(s):  
Creatine Kinase ◽  
Dna Sequences ◽  
Hela Cells ◽  
Lung Tumor ◽  
P53 Protein ◽  
Wild Type Mouse ◽  
High Energy ◽  
Wild Type ◽  
Kinase Gene

The creatine kinases (CK) regenerate ATP for cellular reactions with a high energy expenditure. While muscle CK (CKM) is expressed almost exclusively in adult skeletal and cardiac muscle, brain CK (CKB) expression is more widespread and is highest in brain glial cells. CKB expression is also high in human lung tumor cells, many of which contain mutations in p53 alleles. We have recently detected high levels of CKB mRNA in HeLa cells and, in this study, have tested whether this may be due to the extremely low amounts of p53 protein present in HeLa cells. Transient transfection experiments showed that wild-type mouse p53 severely repressed the rat CKB promoter in HeLa but not CV-1 monkey kidney cells, suggesting that, in HeLa but not CV-1 cells, p53 either associates with a required corepressor or undergoes a posttranslational modification necessary for CKB repression. Conversely, mouse wild-type p53 strongly activated the rat CKM promoter in CV-1 cells but not in HeLa cells, suggesting that, in CV-1 cells, p53 may associate with a required coactivator or is modified in a manner necessary for CKM activation. The DNA sequences required for p53-mediated modulations were found to be within bp -195 to +5 of the CKB promoter and within bp -168 to -97 of the CKM promoter. Moreover, a 112-bp fragment from the proximal rat CKM promoter (bp -168 to -57), which contained five degenerate p53-binding elements, was capable of conferring p53-mediated activation on a heterologous promoter in CV-1 cells. Also, this novel p53 sequence, when situated in the native 168-bp rat CKM promoter, conferred p53-mediated activation equal to or greater than that of the originally characterized far-upstream (bp -3160) mouse CKM p53 element. Therefore, CKB and CKM may be among the few cellular genes which could be targets of p53 in vivo. In addition, we analyzed a series of missense mutants with alterations in conserved region II of p53. Mutations affected p53 transrepression and transactivation activities differently, indicating that these activities in p53 are separable. The ability of p53 mutants to transactivate correlated well with their ability to inhibit transformation of rat embryonic fibroblasts by adenovirus E1a and activated Ras.

Adenylate kinase AK1 knockout heart: energetics and functional performance under ischemia-reperfusion

2002 ◽  
Vol 283 (2) ◽  
pp. H776-H782 ◽  
Author(s):  
Darko Pucar ◽  
Peter Bast ◽  
Richard J. Gumina ◽  
Lynette Lim ◽  
Carmen Drahl ◽  
...  
Keyword(s):  
Adenylate Kinase ◽  
Functional Performance ◽  
Adenine Nucleotide ◽  
Ischemia Reperfusion ◽  
Creatine Phosphate ◽  
Nucleotide Exchange ◽  
Wild Type ◽  
Metabolic Signal ◽  
And Performance ◽  
Ischemic Stress

Deletion of the major adenylate kinase AK1 isoform, which catalyzes adenine nucleotide exchange, disrupts cellular energetic economy and compromises metabolic signal transduction. However, the consequences of deleting the AK1 gene on cardiac energetic dynamics and performance in the setting of ischemia-reperfusion have not been determined. Here, at the onset of ischemia, AK1 knockout mice hearts displayed accelerated loss of contractile force compared with wild-type controls, indicating reduced tolerance to ischemic stress. On reperfusion, AK1 knockout hearts demonstrated reduced nucleotide salvage, resulting in lower ATP, GTP, ADP, and GDP levels and an altered metabolic steady state associated with diminished ATP-to-Pi and creatine phosphate-to-Pi ratios. Postischemic AK1 knockout hearts maintained ∼40% of β-phosphoryl turnover, suggesting increased phosphotransfer flux through remaining adenylate kinase isoforms. This was associated with sustained creatine kinase flux and elevated cellular glucose-6-phosphate levels as the cellular energetic system adapted to deletion of AK1. Such metabolic rearrangements, along with sustained ATP-to-ADP ratio and total ATP turnover rate, maintained postischemic contractile recovery of AK1 knockout hearts at wild-type levels. Thus deletion of the AK1 gene reveals that adenylate kinase phosphotransfer supports myocardial function on initiation of ischemic stress and safeguards intracellular nucleotide pools in postischemic recovery.

scholarly journals Mouse p53 represses the rat brain creatine kinase gene but activates the rat muscle creatine kinase gene.

1994 ◽  
Vol 14 (12) ◽  
pp. 8483-8492 ◽  
Author(s):  
J Zhao ◽  
F I Schmieg ◽  
D T Simmons ◽  
G R Molloy
Keyword(s):  
Creatine Kinase ◽  
Dna Sequences ◽  
Hela Cells ◽  
Lung Tumor ◽  
P53 Protein ◽  
Wild Type Mouse ◽  
High Energy ◽  
Wild Type ◽  
Kinase Gene

The creatine kinases (CK) regenerate ATP for cellular reactions with a high energy expenditure. While muscle CK (CKM) is expressed almost exclusively in adult skeletal and cardiac muscle, brain CK (CKB) expression is more widespread and is highest in brain glial cells. CKB expression is also high in human lung tumor cells, many of which contain mutations in p53 alleles. We have recently detected high levels of CKB mRNA in HeLa cells and, in this study, have tested whether this may be due to the extremely low amounts of p53 protein present in HeLa cells. Transient transfection experiments showed that wild-type mouse p53 severely repressed the rat CKB promoter in HeLa but not CV-1 monkey kidney cells, suggesting that, in HeLa but not CV-1 cells, p53 either associates with a required corepressor or undergoes a posttranslational modification necessary for CKB repression. Conversely, mouse wild-type p53 strongly activated the rat CKM promoter in CV-1 cells but not in HeLa cells, suggesting that, in CV-1 cells, p53 may associate with a required coactivator or is modified in a manner necessary for CKM activation. The DNA sequences required for p53-mediated modulations were found to be within bp -195 to +5 of the CKB promoter and within bp -168 to -97 of the CKM promoter. Moreover, a 112-bp fragment from the proximal rat CKM promoter (bp -168 to -57), which contained five degenerate p53-binding elements, was capable of conferring p53-mediated activation on a heterologous promoter in CV-1 cells. Also, this novel p53 sequence, when situated in the native 168-bp rat CKM promoter, conferred p53-mediated activation equal to or greater than that of the originally characterized far-upstream (bp -3160) mouse CKM p53 element. Therefore, CKB and CKM may be among the few cellular genes which could be targets of p53 in vivo. In addition, we analyzed a series of missense mutants with alterations in conserved region II of p53. Mutations affected p53 transrepression and transactivation activities differently, indicating that these activities in p53 are separable. The ability of p53 mutants to transactivate correlated well with their ability to inhibit transformation of rat embryonic fibroblasts by adenovirus E1a and activated Ras.

Subtle Impact of Akt1 and Akt3 on Exploratory Behavior in Gene Targeted Mice

2015 ◽  
Vol 223 (3) ◽  
pp. 173-180 ◽  
Author(s):  
Christina Leibrock ◽  
Michael Hierlmeier ◽  
Undine E. Lang ◽  
Florian Lang
Keyword(s):  
Forced Swimming Test ◽  
Open Field Test ◽  
Wild Type ◽  
Average Speed ◽  
Closed Area ◽  
Light Area ◽  
Swimming Test ◽  
The Impact ◽  
Center Area ◽  
Dark Transition

Abstract. The present study explored the impact of Akt1 and Akt3 on behavior. Akt1 (akt1-/-) and Akt3 (akt3-/-) knockout mice were compared to wild type (wt) mice. The akt1-/- mice, akt3-/- mice, and wt mice were similar in most parameters of the open-field test. However, the distance traveled in the center area was slightly but significantly less in akt3-/- mice than in wt mice. In the light/dark transition test akt1-/- mice had significantly lower values than wt mice and akt3-/- mice for distance traveled, number of rearings, rearing time in the light area, as well as time spent and distance traveled in the entrance area. They were significantly different from akt3-/- mice in the distance traveled, visits, number of rearings, rearing time in the light area, as well as time spent, distance traveled, number of rearings, and rearing time in the entrance area. In the O-maze the time spent, and the visits to open arms, as well as the number of protected and unprotected headdips were significantly less in akt1-/- mice than in wt mice, whereas the time spent in closed arms was significantly more in akt1-/- mice than in wt mice. Protected and unprotected headdips were significantly less in akt3-/- mice than in wt mice. In closed area, akt3-/- mice traveled a significantly larger distance at larger average speed than akt1-/- mice. No differences were observed between akt1-/- mice, akt3-/- mice and wt-type mice in the time of floating during the forced swimming test. In conclusion, akt1-/- mice and less so akt3-/ mice display subtle changes in behavior.

Slow skeletal muscle myosin-binding protein-C (MyBPC1) mediates recruitment of muscle-type creatine kinase (CK) to myosin

2011 ◽  
Vol 436 (2) ◽  
pp. 437-445 ◽  
Author(s):  
Zhe Chen ◽  
Tong-Jin Zhao ◽  
Jie Li ◽  
Yan-Song Gao ◽  
Fan-Guo Meng ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Creatine Kinase ◽  
Muscle Contraction ◽  
Binding Protein ◽  
High Energy ◽  
Functional Coupling ◽  
Muscle Type ◽  
Myosin Binding Protein ◽  
Slow Skeletal Muscle ◽  
Myosin Binding

Muscle contraction requires high energy fluxes, which are supplied by MM-CK (muscle-type creatine kinase) which couples to the myofibril. However, little is known about the detailed molecular mechanisms of how MM-CK participates in and is regulated during muscle contraction. In the present study, MM-CK is found to physically interact with the slow skeletal muscle-type MyBPC1 (myosin-binding protein C1). The interaction between MyBPC1 and MM-CK depended on the creatine concentration in a dose-dependent manner, but not on ATP, ADP or phosphocreatine. The MyBPC1–CK interaction favoured acidic conditions, and the two molecules dissociated at above pH 7.5. Domain-mapping experiments indicated that MM-CK binds to the C-terminal domains of MyBPC1, which is also the binding site of myosin. The functional coupling of myosin, MyBPC1 and MM-CK is further corroborated using an ATPase activity assay in which ATP expenditure accelerates upon the association of the three proteins, and the apparent Km value of myosin is therefore reduced. The results of the present study suggest that MyBPC1 acts as an adaptor to connect the ATP consumer (myosin) and the regenerator (MM-CK) for efficient energy metabolism and homoeostasis.

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