Abstract
In both humans and mice, the Glu-99-Lys (E99K) mutation in the cardiac actin gene (ACTC) results in little understood apical hypertrophic cardiomyopathy (AHCM). To determine how cross-bridge kinetics change with AHCM development, we applied sinusoidal length perturbations to skinned papillary muscle fibres from 2- and 5-month old E99K transgenic (Tg) and non-transgenic (NTg) mice, and studied tension and its transients. These age groups were chosen because our preliminary studies indicated that AHCM develops with age. Fibres from 5-month old E99K mice showed significant decreases in tension, stiffness, the rate of the medium-speed exponential process and its magnitude compared to non-transgenic control. The nucleotide association constants increased with age, and they were significantly larger in E99K compared to NTg. However, there were no large differences in the rates of the cross-bridge detachment step, the rates of the force generation step, or the phosphate association constant. Our result on force/cross-bridge demonstrates that the decreased active tension of E99K fibres was caused by a decreased amount of force generated per each cross-bridge. The effects were generally less or insignificant at 2 months. A pCa-tension study showed increased Ca2+-sensitivity (pCa50) with age in both the E99K and NTg sample groups, and pCa50 was significantly larger (but only for 0.05–0.06 pCa units) in E99K than in NTg groups. A significant decrease in cooperativity (nH) was observed only in 5-month old E99K mice. We conclude that the AHCM-causing ACTC E99K mutation is associated with progressive alterations in biomechanical parameters, with changes smaller at 2 months but larger at 5 months, correlating with the development of AHCM.
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Abbreviations
- 2πb :
-
Apparent rate constant of the delayed tension (exponential process B)
- 2πc :
-
Apparent rate constant of fast tension recovery (exponential process C)
- ACTA1 :
-
Skeletal actin gene
- ACTC :
-
Cardiac actin gene
- AHCM:
-
Apical HCM
- B :
-
Magnitude of exponential process B
- C :
-
Magnitude of exponential process C
- CK:
-
Creatine kinase
- D :
-
MgADP or its concentration
- E99K:
-
Glu-99-Lys
- f :
-
Frequency of length oscillation
- HCM:
-
Hypertrophic cardiomyopathy
- K 0 :
-
MgADP association constant
- K 1 :
-
MgATP association constant
- k 2 :
-
Rate constant of the cross-bridge detachment step 2
- k − 2 :
-
Rate constant of the reversal of step 2
- K 2 :
-
Equilibrium constant of step 2 (= k2/k− 2)
- k 4 :
-
Rate constant of force generation step 4 (isomerization of the AM.ADP.Pi state)
- k − 4 :
-
Rate constant of the reversal of step 4
- K 4 :
-
Equilibrium constant of step 4 (= k4/k− 4)
- K 5 :
-
Phosphate association constant
- k TR :
-
The rate of tension redevelopment
- LV:
-
Left ventricle
- n H :
-
Cooperativity
- NTg:
-
Non-transgenic
- P :
-
Pi (phosphate) or its concentration
- PCr:
-
Phosphocreatine
- pCa50 :
-
Ca2+ sensitivity
- RV:
-
Right ventricle
- S :
-
MgATP or its concentration
- SCD:
-
Sudden cardiac death
- T 5 :
-
Tension per cross-bridge supported by the AM*ADP.Pi state
- Y(f):
-
Complex modulus
- ΔL :
-
Length change
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Acknowledgements
The authors would like to thank Professor Steven Marston (National Heart and Lung Institute, Imperial College London, London, UK) who developed the Tg mouse model ACTC E99K and made our collaboration possible. The authors also would like to thank Dr. Amy Li in University of Sydney for developing a technique for freezing and thawing muscle samples without causing much damage, and teaching the technique to us. This work was supported by grants from the Natural Science Foundation of Jiangsu Province of China BK20150353 (LW), the National Institutes of Health HL070041 (MK), and The American Heart Association 13GRNT16810043 (MK). The content is solely the responsibility of the authors and does not necessarily reflect the official views of the funding organizations.
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Wang, L., Bai, F., Zhang, Q. et al. Development of apical hypertrophic cardiomyopathy with age in a transgenic mouse model carrying the cardiac actin E99K mutation. J Muscle Res Cell Motil 38, 421–435 (2017). https://doi.org/10.1007/s10974-018-9492-1
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DOI: https://doi.org/10.1007/s10974-018-9492-1