Skeletal muscle dysfunction in muscle-specific LKB1 knockout mice

Department of Physiology and Developmental Biology, 589 WIDB, Brigham Young University, Provo, UT 84602, USA.
Journal of Applied Physiology (Impact Factor: 3.06). 04/2010; 108(6):1775-85. DOI: 10.1152/japplphysiol.01293.2009
Source: PubMed

ABSTRACT Liver kinase B1 (LKB1) is a tumor-suppressing protein that is involved in the regulation of muscle metabolism and growth by phosphorylating and activating AMP-activated protein kinase (AMPK) family members. Here we report the development of a myopathic phenotype in skeletal and cardiac muscle-specific LKB1 knockout (mLKB1-KO) mice. The myopathic phenotype becomes overtly apparent at 30-50 wk of age and is characterized by decreased body weight and a proportional reduction in fast-twitch skeletal muscle weight. The ability to ambulate is compromised with an often complete loss of hindlimb function. Skeletal muscle atrophy is associated with a 50-75% reduction in mammalian target of rapamycin pathway phosphorylation, as well as lower peroxisome proliferator-activated receptor-alpha coactivator-1 content and cAMP response element binding protein phosphorylation (43 and 40% lower in mLKB1-KO mice, respectively). Maximum in situ specific force production is not affected, but fatigue is exaggerated, and relaxation kinetics are slowed in the myopathic mice. The increased fatigue is associated with a 30-78% decrease in mitochondrial protein content, a shift away from type IIA/D toward type IIB muscle fibers, and a tendency (P=0.07) for decreased capillarity in mLKB1-KO muscles. Hearts from myopathic mLKB1-KO mice exhibit grossly dilated atria, suggesting cardiac insufficiency and heart failure, which likely contributes to the phenotype. These findings indicate that LKB1 plays a critical role in the maintenance of both skeletal and cardiac function.

14 Reads
  • Source
    • "Since a role for AMPK in mitochondrial function of skeletal muscle first time was elucidated for more than a decade ago by Winder et al. [30] numerous studies have expanded the knowledge in this field. Use of pharmacological AMPK activators [33], [40], [67]–[70], genetic models [32], [34], [55], [56], [58], [71]–[73] and combination of these [31], [33], [74] have substantiated AMPK as an important mitochondriogenic part. The kinase is involved both in maintenance of basal mitochondrial function and enhancement of mitochondrial enzymes and mitochondrial biogenesis upon increased AMPK activation. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Metformin is used as an anti-diabetic drug. Metformin ameliorates insulin resistance by improving insulin sensitivity in liver and skeletal muscle. Reduced mitochondrial content has been reported in type 2 diabetic muscles and it may contribute to decreased insulin sensitivity characteristic for diabetic muscles. The molecular mechanism behind the effect of metformin is not fully clarified but inhibition of complex I in the mitochondria and also activation of the 5′AMP activated protein kinase (AMPK) has been reported in muscle. Furthermore, both AMPK activation and metformin treatment have been associated with stimulation of mitochondrial function and biogenesis. However, a causal relationship in skeletal muscle has not been investigated. We hypothesized that potential effects of in vivo metformin treatment on mitochondrial function and protein expressions in skeletal muscle are dependent upon AMPK signaling. We investigated this by two weeks of oral metformin treatment of muscle specific kinase dead α2 (KD) AMPK mice and wild type (WT) littermates. We measured mitochondrial respiration and protein activity and expressions of key enzymes involved in mitochondrial carbohydrate and fat metabolism and oxidative phosphorylation. Mitochondrial respiration, HAD and CS activity, PDH and complex I-V and cytochrome c protein expression were all reduced in AMPK KD compared to WT tibialis anterior muscles. Surprisingly, metformin treatment only enhanced respiration in AMPK KD mice and thereby rescued the respiration defect compared to the WT mice. Metformin did not influence protein activities or expressions in either WT or AMPK KD mice. We conclude that two weeks of in vivo metformin treatment enhances mitochondrial respiration in the mitochondrial deficient AMPK KD but not WT mice. The improvement seems to be unrelated to AMPK, and does not involve changes in key mitochondrial proteins.
    PLoS ONE 01/2013; 8(1):e53533. DOI:10.1371/journal.pone.0053533 · 3.23 Impact Factor
  • Source
    • "Subsequent genetic data, however, has shown important roles for LKB1 in muscle. This includes the finding that both skeletal and cardiac muscle phenotypes developed in older LKB1 knockout mice, with decreased voluntary running, type II muscle fiber atrophy, and loss of hind limb muscle function [25]. LKB1 was also shown to affect the differentiation of mouse embryonic fibroblasts (MEFs) into myofibroblasts, contractile cells that express smooth muscle actin and show acto-myosin contractility [26], [27]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Skeletal muscle myoblast differentiation and fusion into multinucleate myotubes is associated with dramatic cytoskeletal changes. We find that microtubules in differentiated myotubes are highly stabilized, but premature microtubule stabilization blocks differentiation. Factors responsible for microtubule destabilization in myoblasts have not been identified. We find that a transient decrease in microtubule stabilization early during myoblast differentiation precedes the ultimate microtubule stabilization seen in differentiated myotubes. We report a role for the serine-threonine kinase LKB1 in both microtubule destabilization and myoblast differentiation. LKB1 overexpression reduced microtubule elongation in a Nocodazole washout assay, and LKB1 RNAi increased it, showing LKB1 destabilizes microtubule assembly in myoblasts. LKB1 levels and activity increased during myoblast differentiation, along with activation of the known LKB1 substrates AMP-activated protein kinase (AMPK) and microtubule affinity regulating kinases (MARKs). LKB1 overexpression accelerated differentiation, whereas RNAi impaired it. Reduced microtubule stability precedes myoblast differentiation and the associated ultimate microtubule stabilization seen in myotubes. LKB1 plays a positive role in microtubule destabilization in myoblasts and in myoblast differentiation. This work suggests a model by which LKB1-induced microtubule destabilization facilitates the cytoskeletal changes required for differentiation. Transient destabilization of microtubules might be a useful strategy for enhancing and/or synchronizing myoblast differentiation.
    PLoS ONE 02/2012; 7(2):e31583. DOI:10.1371/journal.pone.0031583 · 3.23 Impact Factor
  • Source
    • "Given the increased fatigability of KO diaphragms that we show here, and the previously reported finding that voluntary wheel running distance is lower in KO mice, it might be surmised that the apparent shift to IIb MHC is due to decreased overall activity of the KO mice. However, ambulatory activity in young female (Thomson et al., 2010) and male (unpublished results from our laboratory) KO mice is not different than that of C mice. Therefore, the alteration in fiber-type and mitochondrial protein content does not appear to be due to decreased physical activity, assuming that respiratory activity tracks well with ambulation in these mice. "
    [Show abstract] [Hide abstract]
    ABSTRACT: The liver kinase B1 (LKB1)/AMP-activated protein kinase (AMPK) signalling pathway is a major regulator of skeletal muscle metabolic processes. During exercise, LKB1-mediated phosphorylation of AMPK leads to its activation, promoting mitochondrial biogenesis and glucose transport, among other effects. The roles of LKB1 and AMPK have not been fully characterized in the diaphragm. Two methods of AMPK activation were used to characterize LKB1/AMPK signalling in diaphragms from muscle-specific LKB1 knockout (KO) and littermate control mice: (1) acute injection of 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) and (2) 5-min direct electrical stimulation of the diaphragm. Diaphragms were excised 60 min post-AICAR injection and immediately after electrical stimulation. AMPK phosphorylation increased with AICAR and electrical stimulation in control but not KO mice. Acetyl CoA carboxylase phosphorylation increased with AICAR in control but not KO mice, but increased in both genotypes with electrical stimulation. While the majority of mitochondrial protein levels were lower in KO diaphragms, uncoupling protein 3, complex I and cytochrome oxidase IV protein levels were not different between genotypes. KO diaphragms have a lower percentage of IIx fibres and an elevated percentage of IIb fibres when compared with control diaphragms. While in vitro peak force generation was similar between genotypes, KO diaphragms fatigued more quickly and had an impaired ability to recover. LKB1 regulates AMPK phosphorylation, mitochondrial protein expression, fibre type distribution, as well as recovery of the diaphragm from fatigue.
    Acta Physiologica 11/2010; 201(4):457-66. DOI:10.1111/j.1748-1716.2010.02226.x · 4.38 Impact Factor
Show more