scholarly journals In vivo assessment of contractile strength distinguishes differential gene function in skeletal muscle of zebrafish larvae

2015 ◽  
Vol 119 (7) ◽  
pp. 799-806 ◽  
Author(s):  
Brit L. Martin ◽  
Thomas L. Gallagher ◽  
Neha Rastogi ◽  
Jonathan P. Davis ◽  
Christine E. Beattie ◽  
...  

The accessible genetics and extensive skeletal musculature of the zebrafish make it a versatile and increasingly used model for studying muscle contraction. We here describe the development of an in vivo assay for measuring the contractile force of intact zebrafish at the larval stage. In addition, as proof of applicability, we have used this assay to quantify contractile strength of zebrafish larvae in a morphant model of deranged rbfox function. Average maximum tetanic (180 Hz) whole body forces produced by wild-type larvae at 2, 3, 4, and 5 days postfertilization amounted to 3.0, 7.2, 9.1, and 10.8 mN, respectively. To compare at potentially different stages of muscle development, we developed an immunohistological assay for empirically determining the cross-sectional area of larval trunk skeletal muscle to quantify muscle-specific force per cross-sectional area. At 4-5 days postfertilization, specific force amounts to ∼300 mN/mm2, which is similar to fully developed adult mammalian skeletal muscle. We used these assays to measure contractile strength in zebrafish singly or doubly deficient for two rbfox paralogs, rbfox1l and rbfox2, which encode RNA-binding factors shown previously to modulate muscle function and muscle-specific splicing. We found rbfox2 morphants produce maximal tetanic forces similar to wild-type larvae, whereas rbfox1l morphants demonstrate significantly impaired function. rbfox1l/rbfox2 morphants are paralyzed, and their lack of contractile force production in our assay suggests that paralysis is a muscle-autonomous defect. These quantitative functional results allow measurement of muscle-specific phenotypes independent of neural input.

2002 ◽  
Vol 282 (5) ◽  
pp. H1697-H1702 ◽  
Author(s):  
Hui Di Wang ◽  
Douglas G. Johns ◽  
Shanqin Xu ◽  
Richard A. Cohen

Our purpose was to address the role of NAPDH oxidase-derived superoxide anion in the vascular response to ANG II. Blood pressure, aortic superoxide anion, 3-nitrotyrosine, and medial cross-sectional area were compared in wild-type mice and in mice that overexpress human superoxide dismutase (hSOD). The pressor response to ANG II was significantly less in hSOD mice. Superoxide anion levels were increased twofold in ANG II-treated wild-type mice but not in hSOD mice. 3-Nitrotyrosine increased in aortic endothelium and adventitia in wild-type but not hSOD mice. In contrast, aortic medial cross-sectional area increased 50% with ANG II in hSOD mice, comparable to wild-type mice. The lower pressor response to ANG II in the mice expressing hSOD is consistent with a pressor role of superoxide anion in wild-type mice, most likely because it reacts with nitric oxide. Despite preventing the increase in superoxide anion and 3-nitrotyrosine, the aortic hypertrophic response to ANG II in vivo was unaffected by hSOD.


2001 ◽  
Vol 280 (2) ◽  
pp. C288-C295 ◽  
Author(s):  
Robert G. Dennis ◽  
Paul E. Kosnik ◽  
Mark E. Gilbert ◽  
John A. Faulkner

The purpose of this study was to compare the excitability and contractility of three-dimensional skeletal muscle constructs, termed myooids, engineered from C2C12 myoblast and 10T½ fibroblast cell lines, primary muscle cultures from adult C3H mice, and neonatal and adult Sprague-Dawley rats. Myooids were 12 mm long, with diameters of 0.1–1 mm, were excitable by transverse electrical stimulation, and contracted to produce force. After ∼30 days in culture, myooid cross-sectional area, rheobase, chronaxie, resting baseline force, twitch force, time to peak tension, one-half relaxation time, and peak isometric force were measured. Specific force was calculated by dividing peak isometric force by cross-sectional area. The specific force generated by the myooids was 2–8% of that generated by skeletal muscles of control adult rodents. Myooids engineered from C2C12-10T½ cells exhibited greater rheobase, time to peak tension, and one-half relaxation time than myooids engineered from adult rodent cultures, and myooids from C2C12-10T½ and neonatal rat cells had greater resting baseline forces than myooids from adult rodent cultures.


2010 ◽  
Vol 299 (1) ◽  
pp. R268-R276 ◽  
Author(s):  
Anna C. Kayani ◽  
Graeme L. Close ◽  
Wolfgang H. Dillmann ◽  
Ruben Mestril ◽  
Malcolm J. Jackson ◽  
...  

Skeletal muscle atrophy and weakness are major contributors to frailty and impact significantly on quality of life of older people. Muscle aging is characterized by a loss of maximum tetanic force (Po) generation, primarily due to muscle atrophy, to which mitochondrial dysfunction is hypothesized to contribute. We hypothesized that lifelong overexpression of the mitochondrial heat shock protein (HSP) HSP10 in muscle of mice would protect against development of these deficits. Po generation by extensor digitorum longus muscles of adult and old wild-type and HSP10-overexpressing mice was determined in situ. Muscles were subjected to damaging lengthening contractions, and force generation was remeasured at 3 h or 28 days to examine susceptibility to, and recovery from, damage, respectively. Muscles of old wild-type mice had a 23% deficit in Po generation and a 10% deficit in muscle cross-sectional area compared with muscles of adult wild-type mice. Overexpression of HSP10 prevented this age-related fall in Po generation and reduction in cross-sectional area observed in muscles of old wild-type mice. Additionally, overexpression of HSP10 protected against contraction-induced damage independent of age but did not improve recovery if damage occurred. Preservation of muscle force generation and CSA by HSP10 overexpression was associated with protection against the age-related accumulation of protein carbonyls. Data demonstrate that development of age-related muscle weakness may not be inevitable and show, for the first time, that lifelong overexpression of an HSP prevents the age-related loss of Po generation. These findings support the hypothesis that mitochondrial dysfunction is involved in the development of age-related muscle deficits.


2021 ◽  
Vol 12 ◽  
Author(s):  
Rowan R. Smart ◽  
Brian O'Connor ◽  
Jennifer M. Jakobi

Force produced by the muscle during contraction is applied to the tendon and distributed through the cross-sectional area (CSA) of the tendon. This ratio of force to the tendon CSA is quantified as the tendon mechanical property of stress. Stress is traditionally calculated using the resting tendon CSA; however, this does not take into account the reductions in the CSA resulting from tendon elongation during the contraction. It is unknown if calculating the tendon stress using instantaneous CSA during a contraction significantly increases the values of in vivo distal biceps brachii (BB) tendon stress in humans compared to stress calculated with the resting CSA. Nine young (22 ± 1 years) and nine old (76 ± 4 years) males, and eight young females (21 ± 1 years) performed submaximal isometric elbow flexion tracking tasks at force levels ranging from 2.5 to 80% maximal voluntary contraction (MVC). The distal BB tendon CSA was recorded on ultrasound at rest and during the submaximal tracking tasks (instantaneous). Tendon stress was calculated as the ratio of tendon force during contraction to CSA using the resting and instantaneous measures of CSA, and statistically evaluated with multi-level modeling (MLM) and Johnson–Neyman regions of significance tests to determine the specific force levels above which the differences between calculation methods and groups became statistically significant. The tendon CSA was greatest at rest and decreased as the force level increased (p < 0.001), and was largest in young males (23.0 ± 2.90 mm2) followed by old males (20.87 ± 2.0 mm2) and young females (17.08 ± 1.54 mm2) (p < 0.001) at rest and across the submaximal force levels. Tendon stress was greater in the instantaneous compared with the resting CSA condition, and young males had the greatest difference in the values of tendon stress between the two conditions (20 ± 4%), followed by old males (19 ± 5%), and young females (17 ± 5%). The specific force at which the difference between the instantaneous and resting CSA stress values became statistically significant was 2.6, 6.6, and 10% MVC for old males, young females, and young males, respectively. The influence of using the instantaneous compared to resting CSA for tendon stress is sex-specific in young adults, and age-specific in the context of males. The instantaneous CSA should be used to provide a more accurate measure of in vivo tendon stress in humans.


2021 ◽  
Vol 10 (12) ◽  
pp. 2721
Author(s):  
Nobuto Nakanishi ◽  
Shigeaki Inoue ◽  
Rie Tsutsumi ◽  
Yusuke Akimoto ◽  
Yuko Ono ◽  
...  

Ultrasound has become widely used as a means to measure the rectus femoris muscle in the acute and chronic phases of critical illness. Despite its noninvasiveness and accessibility, its accuracy highly depends on the skills of the technician. However, few ultrasound phantoms for the confirmation of its accuracy or to improve technical skills exist. In this study, the authors created a novel phantom model and used it for investigating the accuracy of measurements and for training. Study 1 investigated how various conditions affect ultrasound measurements such as thickness, cross-sectional area, and echogenicity. Study 2 investigated if the phantom can be used for the training of various health care providers in vitro and in vivo. Study 1 showed that thickness, cross-sectional area, and echogenicity were affected by probe compression strength, probe angle, phantom compression, and varying equipment. Study 2 in vitro showed that using the phantom for training improved the accuracy of the measurements taken within the phantom, and Study 2 in vivo showed the phantom training had a short-term effect on improving the measurement accuracy in a human volunteer. The new ultrasound phantom model revealed that various conditions affected ultrasound measurements, and phantom training improved the measurement accuracy.


2005 ◽  
Vol 61 (2) ◽  
Author(s):  
M. A. Gregory ◽  
M. N. Deane ◽  
M. Marsh

Objective: The precise mechanisms by which massage promotes repair in injured soft tissue are unknown. Various authorshave attributed the beneficial effects of massage to vasodilation and increased skin and muscle blood flow. The aim of this study was to determine whether deep transverse friction massage (DTF) causes capillary vasodilation in untraumatised skeletal muscle. Setting: Academic institution.Interventions: Twelve New Zealand white rabbits were anaesthetised and the left biceps femoris muscle received 10 minutes of DTF. Following treatment, wedge biopsies were taken from the musclewithin 10 minutes of treatment (R1 - 4), 24 hours (R5 - 8) and 6 days(R9 - 12) after treatment. To serve as controls, similar biopsies weretaken from the right biceps femoris of animals. The samples were fixed, dehydrated and embedded in epoxy resin.Transverse sections (1µm) of muscle were cut, stained with 1% aqueous alkaline toluidine blue and examined with a light microscope using a 40X objective. Images containing capillaries were captured using an image analyser with SIS software and the cross sectional diameters of at least 60 capillaries were measured from each specimen. Main Outcome Measures: Changes in capillary diameter. Results: The mean capillary diameters in control muscle averaged 4.76 µm. DTF caused a significant immediate increase of 17.3% in cross sectional area (p<0.001), which was not significantly increased by 10.0% after 24 hours (p>0.05). Six days after treatment the cross-sectional area of the treated muscle was 7.6% smaller than the controls. Conclusions: This confirms the contention that DTF stimulates muscle blood flow immediately after treatment and this may account for its beneficial effects in certain conditions. 


1994 ◽  
Vol 77 (2) ◽  
pp. 947-955 ◽  
Author(s):  
M. I. Lewis ◽  
S. A. Monn ◽  
W. Z. Zhan ◽  
G. C. Sieck

Interactive effects of emphysema (EMP) and prolonged nutritional deprivation (ND) on contractile, morphometric, and metabolic properties of hamster diaphragm muscle (DIA) were examined. Six months after induction of EMP (intratracheal elastase), saline-treated controls (CTL) and EMP hamsters of similar body weights were subjected to ND over 6 wk. Isometric contractile and fatigue properties of costal DIA were determined in vitro. DIA fibers were histochemically classified as type I or II, and fiber succinate dehydrogenase activity and cross-sectional area were determined using quantitative microscopic procedures. From histochemical sections, the number of capillaries per fiber (C/F) and per fiber cross-sectional area (C/A) were determined. ND resulted in progressive loss of body weight (ND-CTL, 23.8%; ND-EMP, 28.4%; P = NS). ND did not affect reduction in optimal length (Lo) of DIA fibers in EMP compared with CTL and ND-CTL hamsters. Maximum specific force (i.e., force/unit area) was reduced by approximately 25% in EMP animals compared with CTL. ND did not improve or exacerbate the reduction in specific force with EMP. ND attenuated improved fatigue resistance of DIA in EMP animals. No differences in fiber type proportions were noted among experimental groups. Significant atrophy of type I and II DIA fibers was noted after ND. Atrophy was proportionately greater in type II fibers of ND-EMP when referenced to EMP animals. Thus adaptive hypertrophy of type II DIA fibers in EMP animals was abolished. Fiber succinate dehydrogenase activity was significantly increased in type I and II fibers in EMP DIA. ND did not affect this metabolic adaptation of DIA fibers to persistent loads imposed by EMP.(ABSTRACT TRUNCATED AT 250 WORDS)


1992 ◽  
Vol 73 (3) ◽  
pp. 1165-1170 ◽  
Author(s):  
J. D. MacDougall ◽  
C. E. Webber ◽  
J. Martin ◽  
S. Ormerod ◽  
A. Chesley ◽  
...  

Our purpose was to investigate the relationship between running volume and bone mineral mass in adult male runners. Whole body and regional bone mineral density were determined by dual-photon absorptiometry in 22 sedentary controls and 53 runners who were selected according to their running mileage to fall into a 5- to 10-, 15- to 20-, 25- to 30-, 40- to 55-, or 60- to 75-mile/wk group. All groups were of similar age (20–45 yr) and nutritional status, as determined by 7-day food records. Regional sites for bone density measurements included the trunk, spine, pelvis, thighs, and lower legs. In addition, serum total testosterone was determined in each subject and computed tomography scans were made of the lower legs in 34 subjects to assess bone cross-sectional area. No significant differences were detected for bone density measurements with the exception of the lower legs where it was significantly (P less than 0.05) greater for the 15- to 20-mile/wk group than for the control and 5- to 10-mile/wk groups. With mileage greater than 20 miles/wk, bone density of the lower legs showed no further increase and, in fact, tended to decrease, so that for the 60- to 75-mile/wk group it was similar to that of the controls. Cross-sectional area of the tibia and fibula when normalized to body weight tended to be greater as weekly mileage increased and was significantly greater in the 40- to 55-mile/wk runners than in the control group.(ABSTRACT TRUNCATED AT 250 WORDS)


2012 ◽  
Vol 303 (6) ◽  
pp. L519-L527 ◽  
Author(s):  
Vladimir T. Basic ◽  
Elsa Tadele ◽  
Ali Ateia Elmabsout ◽  
Hongwei Yao ◽  
Irfan Rahman ◽  
...  

Cigarette smoke (CS) is a well-established risk factor in the development of chronic obstructive pulmonary disease (COPD). In contrast, the extent to which CS exposure contributes to the development of the systemic manifestations of COPD, such as skeletal muscle dysfunction and wasting, remains largely unknown. Decreased skeletal muscle capillarization has been previously reported in early stages of COPD and might play an important role in the development of COPD-associated skeletal muscle abnormalities. To investigate the effects of chronic CS exposure on skeletal muscle capillarization and exercise tolerance, a mouse model of CS exposure was used. The 129/SvJ mice were exposed to CS for 6 mo, and the expression of putative elements of the hypoxia-angiogenic signaling cascade as well as muscle capillarization were studied. Additionally, functional tests assessing exercise tolerance/endurance were performed in mice. Compared with controls, skeletal muscles from CS-exposed mice exhibited significantly enhanced expression of von Hippel-Lindau tumor suppressor (VHL), ubiquitin-conjugating enzyme E2D1 (UBE2D1), and prolyl hydroxylase-2 (PHD2). In contrast, hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) expression was reduced. Furthermore, reduced muscle fiber cross-sectional area, decreased skeletal muscle capillarization, and reduced exercise tolerance were also observed in CS-exposed animals. Taken together, the current results provide evidence linking chronic CS exposure and induction of VHL expression in skeletal muscles leading toward impaired hypoxia-angiogenesis signal transduction, reduced muscle fiber cross-sectional area, and decreased exercise tolerance.


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