Short telomeres and stem cell exhaustion model Duchenne muscular dystrophy in mdx/mTR mice.

In Duchenne muscular dystrophy (DMD), dystrophin mutation leads to progressive lethal skeletal muscle degeneration. For unknown reasons, dystrophin deficiency does not recapitulate DMD in mice (mdx), which have mild skeletal muscle defects and potent regenerative capacity. We postulated that human DMD progression is a consequence of loss of functional muscle stem cells (MuSC), and the mild mouse mdx phenotype results from greater MuSC reserve fueled by longer telomeres. We report that mdx mice lacking the RNA component of telomerase (mdx/mTR) have shortened telomeres in muscle cells and severe muscular dystrophy that progressively worsens with age. Muscle wasting severity parallels a decline in MuSC regenerative capacity and is ameliorated histologically by transplantation of wild-type MuSC. These data show that DMD progression results, in part, from a cell-autonomous failure of MuSC to maintain the damage-repair cycle initiated by dystrophin deficiency. The essential role of MuSC function has therapeutic implications for DMD.

Minimally invasive high-speed imaging of sarcomere contractile dynamics in mice and humans.

Sarcomeres are the basic contractile units of striated muscle. Our knowledge about sarcomere dynamics has primarily come from in vitro studies of muscle fibres and analysis of optical diffraction patterns obtained from living muscles. Both approaches involve highly invasive procedures and neither allows examination of individual sarcomeres in live subjects. Here we report direct visualization of individual sarcomeres and their dynamical length variations using minimally invasive optical microendoscopy to observe second-harmonic frequencies of light generated in the muscle fibres of live mice and humans. Using microendoscopes as small as 350 microm in diameter, we imaged individual sarcomeres in both passive and activated muscle. Our measurements permit in vivo characterization of sarcomere length changes that occur with alterations in body posture and visualization of local variations in sarcomere length not apparent in aggregate length determinations. High-speed data acquisition enabled observation of sarcomere contractile dynamics with millisecond-scale resolution. These experiments point the way to in vivo imaging studies demonstrating how sarcomere performance varies with physical conditioning and physiological state, as well as imaging diagnostics revealing how neuromuscular diseases affect contractile dynamics.

The sonographic "bright band sign" of splenic infarction.

OBJECTIVES: To evaluate the frequency of the "bright band sign" in patients with splenic infarcts as well as control patients and to thereby assess whether the bright band sign has potential utility as a sonographic sign of splenic infarction. METHODS: Using an electronic search engine and image review, 37 patients were retrospectively identified with noncystic parenchymal splenic infarcts on sonography. Nineteen abnormal control patients with noninfarcted splenic lesions on sonography and 100 normal control patients with sonographically normal spleens were also identified. The sonographic appearance of each splenic lesion was evaluated by 2 reviewers and assessed for the bright band sign, defined as thin specular reflectors perpendicular to the sound beam within hypoechoic parenchymal lesions, and for the presence or absence of the classic sonographic appearance of splenic infarction. Possible histologic counterparts of the bright band sign were assessed in archival infarct specimens. RESULTS: The bright band sign was present in 34 (91.9%; 95% confidence interval [CI], 78.1%-98.3%) of 37 patients with splenic infarcts on sonography, including 12 (85.7%; 95% CI, 57.2%-98.2%) of 14 with classic and 22 (95.7%; 95% CI, 78.1%-99.9%) of 23 with nonclassic infarct appearances. No normal or abnormal control patients had the bright band sign. Histologic sections suggested that preserved splenic trabeculae within infarcts may generate the bright band sign. CONCLUSIONS: The bright band sign is a potentially useful sonographic sign of splenic infarction, which may confer additional sensitivity and specificity and may be particularly helpful with infarcts having nonclassic appearances.

Chronic exertional compartment syndrome: muscle changes with isometric exercise.

UNLABELLED: Chronic exertional compartment syndrome (CECS) is a well-documented cause of lower leg pain in active individuals. The pathophysiology is unclear, although it is generally believed to be associated with increased intramuscular pressure, but there is very little information about muscle function in relation to the onset of pain. PURPOSE: To investigate strength, fatigue, and recovery of the anterior tibial muscles in CECS patients and healthy subjects during an isometric exercise protocol. METHODS: Twenty patients and 22 control subjects (mean age 27.6 yr and 33.0 yr, respectively) performed a 20-min isometric exercise protocol consisting of intermittent maximal voluntary contractions (MVC). Central fatigue was evaluated by comparing changes in electrically stimulated (2 s at 50 Hz) and voluntary contraction force before and during the exercise, and then throughout 10 min of recovery. Muscle size was measured by ultrasonography. Pain and cardiovascular parameters were also examined. RESULTS: The absolute MVC forces were similar, but MVC:body mass of the patients was lower (P < 0.05) as was the ratio of MVC to muscle cross-sectional area (P < 0.01). The extent of central and peripheral fatigue was similar in the two groups. The patients reported significantly higher levels of pain during exercise (P < 0.05 at 4 min) and after the first minute of recovery (P < 0.001). An 8% increase in muscle size after exercise was observed for both groups. There were no differences in the cardiovascular responses of the two groups. CONCLUSIONS: CECS patients were somewhat weaker than normal but fatigued at a similar rate during isometric exercise. Patients reported higher pain than controls despite comparable changes in muscle size, suggesting that abnormally tight fascia are not the main cause of CECS symptoms.

Rejuvenation of the muscle stem cell population restores strength to injured aged muscles.

The elderly often suffer from progressive muscle weakness and regenerative failure. We demonstrate that muscle regeneration is impaired with aging owing in part to a cell-autonomous functional decline in skeletal muscle stem cells (MuSCs). Two-thirds of MuSCs from aged mice are intrinsically defective relative to MuSCs from young mice, with reduced capacity to repair myofibers and repopulate the stem cell reservoir in vivo following transplantation. This deficiency is correlated with a higher incidence of cells that express senescence markers and is due to elevated activity of the p38α and p38ß mitogen-activated kinase pathway. We show that these limitations cannot be overcome by transplantation into the microenvironment of young recipient muscles. In contrast, subjecting the MuSC population from aged mice to transient inhibition of p38α and p38ß in conjunction with culture on soft hydrogel substrates rapidly expands the residual functional MuSC population from aged mice, rejuvenating its potential for regeneration and serial transplantation as well as strengthening of damaged muscles of aged mice. These findings reveal a synergy between biophysical and biochemical cues that provides a paradigm for a localized autologous muscle stem cell therapy for the elderly.

Orderly recruitment of motor units under optical control in vivo.

A drawback of electrical stimulation for muscle control is that large, fatigable motor units are preferentially recruited before smaller motor units by the lowest-intensity electrical cuff stimulation. This phenomenon limits therapeutic applications because it is precisely the opposite of the normal physiological (orderly) recruitment pattern; therefore, a mechanism to achieve orderly recruitment has been a long-sought goal in physiology, medicine and engineering. Here we demonstrate a technology for reliable orderly recruitment in vivo. We find that under optical control with microbial opsins, recruitment of motor units proceeds in the physiological recruitment sequence, as indicated by multiple independent measures of motor unit recruitment including conduction latency, contraction and relaxation times, stimulation threshold and fatigue. As a result, we observed enhanced performance and reduced fatigue in vivo. These findings point to an unanticipated new modality of neural control with broad implications for nervous system and neuromuscular physiology, disease research and therapeutic innovation.