Elsevier

Joint Bone Spine

Volume 75, Issue 1, January 2008, Pages 53-57
Joint Bone Spine

Original article
Ubiquitin ligases MuRF1 and MAFbx in human skeletal muscle atrophy

https://doi.org/10.1016/j.jbspin.2007.04.019Get rights and content

Abstract

Introduction

Several pathological conditions can induce skeletal muscle atrophy and seem to share common enzyme pathways. In catabolic states where proteolysis is increased, two genes specific to muscle atrophy, MuRf1 and MAFbx, are upregulated. These encode ubiquitin ligases, which bind to and mediate ubiquitination of myofibrillar proteins for subsequent degradation during muscle atrophy.

Methods

Fifteen patients undergoing leg amputation were divided into two groups. Group A included 12 elderly patients (mean age 79 years) amputated for vascular disease (complicated by diabetes in four), chronic osteomyelitis or squamous cell carcinoma. Group B included three car accident victims (mean age 32 years) amputated due to acute arterial insufficiency. Gastrocnemius muscle biopsies were collected for a histochemical and immunohistochemical (anti-MuRf1, anti-MAFbx) study.

Results

Group A specimens showed a decreased cross-sectional fiber area and length, adipose tissue replacement, and MuRf1 and MAFbx immunoreactivity. Muscle cells showed MuRf1 and MAFbx subsarcolemmal immunoreactivity and weak extracellular matrix immunoreactivity. Group B samples exhibited mild muscle structural changes; they did not stain with anti-MuRf1 or anti-MAFbx, and neither did sections showing muscle degeneration and adipose tissue replacement.

Discussion

Results of our preliminary study showed upregulation of MuRf1 and MAFbx in atrophied muscle and support their role as regulatory peptides in various conditions that lead to muscle atrophy. Data suggest that the study of cellular pathways can help identify promising targets for effective new treatments for skeletal muscle atrophy.

Conclusion

The treatment of several orthopedic conditions is complicated by muscle atrophy; potential treatments could be directed to specific sites where these proteins are localized.

Introduction

The basic definition of skeletal muscle atrophy is a decrease in cell size. Atrophy can be due to a decrease in the fiber cross-sectional area or length, or both, and can also occur as a decrease in muscle mass, protein content or decreased fiber number, and loss of strength [1]. Numerous and heterogeneous pathological conditions cause human muscle atrophy [2]. Profound atrophy is often a consequence of diseases such as cancer and AIDS. Muscle immobilization, as commonly seen when a limb is placed in a cast after an orthopedic injury, causes rapid muscle loss that may require months of physical therapy to reverse. The effectiveness of glucocorticoid drugs, such as dexamethasone, is limited by muscle wasting, seen as a side-effect of these agents. Even during normal aging, there is a gradual loss of muscle mass and a diminished capacity to reverse that loss, which results in weakness and morbidity [3], [4], [5], [6], [7]. Experimental studies have proven that different conditions use common enzyme pathways, leading to muscle atrophy [5]. The primary mechanism that contributes to decreasing skeletal muscle strength and size in healthy aging is not presently known [8].

In catabolic conditions where proteolysis is increased, two specific genes for skeletal muscle atrophy, MuRf1 (muscle ring finger-1) and MAFbx (muscle atrophy F-box) (or atrogin-1), which encode ubiquitin (Ub) ligases, are upregulated. Ligases bind to and mediate ubiquitination of myofibrillar proteins for subsequent degradation during muscle atrophy and are considered as sensitive markers of muscle protein degradation by the Ub-proteasome system [9], [10].

Since there are no effective treatments for muscle atrophy, these proteins are potential targets for the pharmacological treatment of muscle atrophy [11]. We focused on similarities in the intracellular responses of human skeletal muscle atrophy by including subjects in whom a variety of diverse stimuli induce muscle atrophy. The aim of the preliminary study was to gain a better understanding of the pathological anatomy of human lower limb muscles in atrophy caused by different pathological conditions by use of histochemistry and immunolocalization of MuRF1 and MAFbx and showing that MuRF1 and MAFbx are not expressed in normal skeletal muscle.

Section snippets

Methods

Fifteen patients undergoing amputation in the distal or proximal third of the lower leg due to diverse chronic or acute conditions from March to September 2005 were divided into two groups. Group A included 12 elderly subjects (mean age 79 years, range 65–85), of whom 10 underwent amputation for vascular disease (four from complications of type 2 diabetes), one for chronic osteomyelitis four years after sustaining an exposed fracture, and one for squamous cell carcinoma of the skin on the

Histochemistry

Relationships between muscle-tendon junctions (MTJs), myofascial junctions and myofiber-myofiber (myomuscular) junctions were clearly apparent. In Group A, all specimens presented split fibers and fibers with centralized nuclei; fiber atrophy and decreased cross-sectional fiber area and length, with the presence of adipose tissue, were also observed. Group B muscle fibers had a normal appearance, with centralized nuclei.

Immunohistochemistry

Group A specimens stained for anti-MuRf1 (Fig. 1a,c; Table 1) and

Discussion

Skeletal muscle atrophy occurs as a consequence of aging, denervation, injury, joint immobilization, bed rest, glucocorticoid treatment, sepsis and cancer. Although a variety of diverse stimuli induce muscle atrophy, there is a surprising number of similarities in intracellular responses [12], [13], [14], [15], [16], [17]. Bodine and co-workers performed transcript profiling to identify candidate molecular mediators of muscle atrophy. Although several genes were upregulated in a rat model of

Acknowledgments

The authors are grateful to Ms Sandra Manzotti of the Laboratory of the Department of Orthopedics, Università Politecnica delle Marche, for the preparation of the histological sections and to Dr Silvia Modena for reviewing the English.

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