RESUMEN
The capability of regeneration for skeletal muscle after injury depends on the differentiation and proliferation ability of the resident stem cells called satellite cells. It has been reported that electrical stimulation was widely used in clinical conditions to facilitate muscle regeneration after injury, but the characterization of satellite cell responses to the context of low-frequency electrical stimulation in early-phase muscle strain conditions has not been fully clarified. In this study, we aim to investigate the effects of low-frequency electrical stimulation (frequency: 20 Hz; duration: 30 minutes, twice daily) on satellite cell activities in a rat model for the early phase of muscle strain. Firstly, we adopted our previously developed rat model to mimic the early phase of muscle strain in human. After then, we examined the effects of low-frequency electrical stimulation on histopathological changes of the muscle fiber by hematoxylin and eosin (H&E) staining. Finally, we investigated the effects of low-frequency electrical stimulation on satellite cell proliferation and differentiation by quantification of the expression level of the specific proteins using western blot analyses. The muscle strain in biceps femoris muscles of rats can be induced by high-speed rotation from knee flexion 50° to full knee extension at 960°·s-1 angular velocity during its tetany by activating the sciatic nerve, as evidenced by a widening of the interstitial space between fibers, and more edema or necrosis fibers were detected in the model rats without treatment than in control rats. After treatment with low-frequency electrical stimulation (frequency: 20 Hz; duration: 30 minutes, twice daily), the acute strained biceps femoris muscles of rats showed obvious improvement of histomorphology as indicated by more mature muscle fibers with well-ordered formation with clear boundaries. Consistently, the expression levels of the MyoD and myogenin were marked higher than those in the rats in the animal model group, indicating increased satellite cell proliferating and differentiating activities by low-frequency electrical stimulation. This study shows that low-frequency electrical stimulation provides an effective stimulus to upregulate the protein expression of MyoD/myogenin and accelerate the restoration of structure during the early phase of muscle strain. This may have significance for clinical practice. Optimization of low-frequency electrical stimulation parameters may enhance the therapeutic outcome in patients.
Asunto(s)
Estimulación Eléctrica , Fibras Musculares Esqueléticas , Regeneración/efectos de la radiación , Células Satélite del Músculo Esquelético , Animales , Masculino , Fibras Musculares Esqueléticas/citología , Fibras Musculares Esqueléticas/patología , Fibras Musculares Esqueléticas/fisiología , Fibras Musculares Esqueléticas/efectos de la radiación , Proteína MioD/metabolismo , Miogenina/metabolismo , Ratas , Ratas Sprague-Dawley , Células Satélite del Músculo Esquelético/citología , Células Satélite del Músculo Esquelético/fisiología , Células Satélite del Músculo Esquelético/efectos de la radiaciónRESUMEN
Skeletal muscle stem (satellite) cells transplanted into host mouse muscles contribute to muscle regeneration. Irradiation of host muscle enhances donor stem cell engraftment by promoting the proliferation of transplanted donor cells. We hypothesised that, similar to other systems, cells damaged by radiation might be effecting this donor cell proliferation. But we found no difference in the percentage of dying (TUNEL+) cells in immunodeficient dystrophic mouse muscles at the times after the irradiation dose that enhances donor cell engraftment. Similarly, irradiation did not significantly increase the number of TUNEL+ cells in non-dystrophic immunodeficient mouse muscles and it only slightly enhanced donor satellite cell engraftment in this mouse strain, suggesting either that the effector cells are present in greater numbers within dystrophic muscle, or that an innate immune response is required for effective donor cell engraftment. Donor cell engraftment within non-irradiated dystrophic host mouse muscles was not enhanced if they were transplanted with either satellite cells, or myofibres, derived from irradiated dystrophic mouse muscle. But a mixture of cells from irradiated muscle transplanted with donor satellite cells promoted donor cell engraftment in a few instances, suggesting that a rare, yet to be identified, cell type within irradiated dystrophic muscle enhances the donor stem cell-mediated regeneration. The mechanism by which cells within irradiated host muscle promote donor cell engraftment remains elusive.
Asunto(s)
Inflamación/terapia , Músculo Esquelético/citología , Músculo Esquelético/efectos de la radiación , Distrofia Muscular Animal/terapia , Regeneración , Células Satélite del Músculo Esquelético/efectos de la radiación , Células Satélite del Músculo Esquelético/trasplante , Animales , Diferenciación Celular , Proliferación Celular , Femenino , Rayos gamma , Inflamación/inmunología , Masculino , Ratones , Ratones Endogámicos mdx , Distrofia Muscular Animal/inmunología , Células Satélite del Músculo Esquelético/inmunologíaRESUMEN
To study the role of IGF-1 on stimulation with monochromatic light during incubation altering posthatch muscle growth, chicken embryos were exposed to blue light, green light, red light, white light or darkness throughout embryonic period and then were raised in white light conditions upon hatching. Comparing with the other treatment groups, the chicks in green light group had heavier hatching weights, higher muscle indexes and larger muscle fibers. Both in vivo and in vitro studies showed that the number and proliferative activity of satellite cells in green light group were the highest. Plasma IGF-1 level and skeletal muscle IGF-1R mRNA level were higher in green light group. Moreover, exogenous IGF-1 increased the proliferative activity of satellite cell in a dose-dependent fashion. These results suggest that stimulation with monochromatic green light during incubation promoted posthatch muscle growth and satellite cell proliferation of chicks through IGF-1 signaling.
Asunto(s)
Proliferación Celular/efectos de la radiación , Factor I del Crecimiento Similar a la Insulina/metabolismo , Luz , Desarrollo de Músculos/efectos de la radiación , Células Satélite del Músculo Esquelético/efectos de la radiación , Animales , Embrión de Pollo , Pollos , Receptor IGF Tipo 1/metabolismo , Células Satélite del Músculo Esquelético/metabolismo , Células Satélite del Músculo Esquelético/fisiología , Transducción de SeñalRESUMEN
There is fear that mechanical overloading (OVL; ie, high-force contractions) accelerates Duchenne muscular dystrophy. Herein, we determined whether short-term OVL combined with wheel running, short-term OVL combined with irradiation, and long-term OVL are detrimental for hind limb mdx mouse muscle, a murine model of Duchene muscular dystrophy exhibiting milder dystrophic features. OVL was induced by the surgical ablation of the synergic muscles of the plantaris muscle, a fast muscle susceptible to contraction-induced muscle damage in mdx mice. We found that short-term OVL combined with wheel and long-term OVL did not worsen the deficit in specific maximal force (ie, absolute maximal force normalized to muscle size) and histological markers of muscle damage (percentage of regenerating fibers and fibrosis) in mdx mice. Moreover, long-term OVL did not increase the alteration in calcium homeostasis and did not deplete muscle cell progenitors expressing Pax 7 in mdx mice. Irradiation before short-term OVL, which is believed to inhibit muscle regeneration, was not more detrimental to mdx than control mice. Interestingly, short-term OVL combined with wheel and long-term OVL markedly improved the susceptibility to contraction-induced damage, increased absolute maximal force, induced hypertrophy, and promoted a slower, more oxidative phenotype. Together, these findings indicate that OVL is beneficial to mdx muscle, and muscle regeneration does not mask the potentially detrimental effect of OVL.
Asunto(s)
Músculo Esquelético/fisiopatología , Distrofia Muscular Animal/fisiopatología , Distrofia Muscular de Duchenne/fisiopatología , Animales , Modelos Animales de Enfermedad , Femenino , Hipertrofia , Extremidad Inferior , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos mdx , Actividad Motora , Contracción Muscular , Músculo Esquelético/efectos de la radiación , Mutación , Regeneración , Células Satélite del Músculo Esquelético/fisiología , Células Satélite del Músculo Esquelético/efectos de la radiaciónRESUMEN
The loss of genome integrity in adult stem cells results in accelerated tissue aging and is possibly cancerogenic. Adult stem cells in different tissues appear to react robustly to DNA damage. We report that adult skeletal stem (satellite) cells do not primarily respond to radiation-induced DNA double-strand breaks (DSBs) via differentiation and exhibit less apoptosis compared to other myogenic cells. Satellite cells repair these DNA lesions more efficiently than their committed progeny. Importantly, non-proliferating satellite cells and post-mitotic nuclei in the fiber exhibit dramatically distinct repair efficiencies. Altogether, reduction of the repair capacity appears to be more a function of differentiation than of the proliferation status of the muscle cell. Notably, satellite cells retain a high efficiency of DSB repair also when isolated from the natural niche. Finally, we show that repair of DSB substrates is not only very efficient but, surprisingly, also very accurate in satellite cells and that accurate repair depends on the key non-homologous end-joining factor DNA-PKcs.
Asunto(s)
Reparación del ADN por Unión de Extremidades , Músculo Esquelético/citología , Animales , Apoptosis/efectos de la radiación , Células Cultivadas , Roturas del ADN de Doble Cadena/efectos de la radiación , Citometría de Flujo , Rayos gamma , Histonas/metabolismo , Ratones , Mioblastos/citología , Mioblastos/efectos de los fármacos , Células Satélite del Músculo Esquelético/citología , Células Satélite del Músculo Esquelético/efectos de la radiación , Células Madre/citología , Células Madre/metabolismo , Células Madre/efectos de la radiaciónRESUMEN
Tissue engineering represents a promising research field, targeting the creation of new functional muscle tissue in vitro. The aim of the present study was to show the influence of static magnetic fields (SMF) and insulin-like growth factor-1 (IGF1), as enhancing stimuli on human satellite cell cultures, which are preferred sources of stem cells in engineering skeletal muscle tissue. To detect effects on myogenic maturation and proliferation, AlamarBlue® proliferation, assay and semi-quantitative reverse transcription-polymerase chain reaction of following markers was performed: desmin (DES), myogenic factor-5 (MYF5), myogenic differentiation antigen-1 (MYOD1), myogenin (MYOG), myosin heavy chain (MYH) and α1 actin (ACTA1). As a distinct marker of differentiation, immunohistochemical staining and fusion index determination was performed on satellite cell cultures stimulated with IGF1 and IGF1-plus-SMF with an intensity of 80 mT. Proliferation was increased by additional SMF application to IGF1-stimulated cell cultures on the first day of myogenesis. Relative gene expression of measured markers was increased by IGF1 application in the first days of myogenesis except for ACTA1. Additional SMF application enhanced this effect. Nevertheless we were unable to demonstrate the formation of contractile muscle tissue. Immunhistochemical staining verified muscle origin and all markers were displayed.
Asunto(s)
Factor I del Crecimiento Similar a la Insulina/farmacología , Campos Magnéticos , Células Satélite del Músculo Esquelético/efectos de los fármacos , Células Satélite del Músculo Esquelético/efectos de la radiación , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/efectos de la radiación , Proliferación Celular/efectos de los fármacos , Proliferación Celular/efectos de la radiación , Células Cultivadas , Perfilación de la Expresión Génica , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/efectos de la radiación , Humanos , Células Satélite del Músculo Esquelético/citología , Células Satélite del Músculo Esquelético/metabolismoRESUMEN
Tissue engineering is a promising research field, which aims to create new functional muscle tissue in vitro, by utilizing the myogenic differentiation potential of human stem cells. The objective of the present study was to determine the effect of static magnetic fields (SMF), combined with the use of the myogenic differentiation enhancing hepatocyte growth factor (HGF), on human satellite cell cultures, which are one of the preferred stem cell sources in skeletal muscle tissue engineering. We performed almarBlue® proliferation assays and semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) for the following myogenic markers: desmin (DES), myogenic factor 5 (MYF5), myogenic differentiation antigen 1 (MYOD1), myogenin (MYOG), myosin heavy chain (MYH) and α1 actin (ACTA1) to detect the effects on myogenic maturation. Additionally, immunohistochemical staining (ICC) and fusion index (FI) determination as independent markers of differentiation were performed on satellite cell cultures stimulated with HGF and HGF + SMF with an intensity of 80 mT. ICC verified the muscle phenotype at all time points. SMF enhanced the proliferation of satellite cell cultures treated with HGF. RT-PCR analysis, ICC and FI calculation revealed the effects of HGF/SMF on the investigated differentiation markers and stimulation with HGF and SMF verified the continuing maturation, however no significant increase in analysed markers could be detected when compared with control cultures treated with serum cessation. In conclusion, HGF or HGF + SMF stimulation of human satellite cell cultures did not lead to the desired enhancement of myogenic maturation of human satellite cell cultures compared with cell cultures stimulated with growth factor reduction.
Asunto(s)
Factor de Crecimiento de Hepatocito/farmacología , Campos Magnéticos , Células Satélite del Músculo Esquelético/efectos de los fármacos , Células Satélite del Músculo Esquelético/efectos de la radiación , Actinas/genética , Actinas/metabolismo , Proliferación Celular/efectos de los fármacos , Proliferación Celular/efectos de la radiación , Células Cultivadas , Desmina/genética , Desmina/metabolismo , Expresión Génica , Humanos , Proteína MioD/genética , Proteína MioD/metabolismo , Factor 5 Regulador Miogénico/genética , Factor 5 Regulador Miogénico/metabolismo , Miogenina/genética , Miogenina/metabolismo , Células Satélite del Músculo Esquelético/metabolismoRESUMEN
Extraocular muscles (EOM) have a strikingly different disease profile than limb skeletal muscles. It has long been known that they are spared in Duchenne (DMD) and other forms of muscular dystrophy. Despite many studies, the cause for this sparing is not understood. We have proposed that differences in myogenic precursor cell properties in EOM maintain normal morphology over the lifetime of individuals with DMD due to either greater proliferative potential or greater resistance to injury. This hypothesis was tested by exposing wild type and mdx:utrophin(+/-) (het) mouse EOM and limb skeletal muscles to 18 Gy gamma irradiation, a dose known to inhibit satellite cell proliferation in limb muscles. As expected, over time het limb skeletal muscles displayed reduced central nucleation mirrored by a reduction in Pax7-positive cells, demonstrating a significant loss in regenerative potential. In contrast, in the first month post-irradiation in the het EOM, myofiber cross-sectional areas first decreased, then increased, but ultimately returned to normal compared to non-irradiated het EOM. Central nucleation significantly increased in the first post-irradiation month, resembling the dystrophic limb phenotype. This correlated with decreased EECD34 stem cells and a concomitant increase and subsequent return to normalcy of both Pax7 and Pitx2-positive cell density. By two months, normal het EOM morphology returned. It appears that irradiation disrupts the normal method of EOM remodeling, which react paradoxically to produce increased numbers of myogenic precursor cells. This suggests that the EOM contain myogenic precursor cell types resistant to 18 Gy gamma irradiation, allowing return to normal morphology 2 months post-irradiation. This supports our hypothesis that ongoing proliferation of specialized regenerative populations in the het EOM actively maintains normal EOM morphology in DMD. Ongoing studies are working to define the differences in the myogenic precursor cells in EOM as well as the cellular milieu in which they reside.
Asunto(s)
Rayos gamma/efectos adversos , Distrofia Muscular Animal/patología , Músculos Oculomotores/patología , Músculos Oculomotores/efectos de la radiación , Utrofina/metabolismo , Animales , Proliferación Celular/efectos de la radiación , Extremidades/patología , Extremidades/efectos de la radiación , Proteínas de Homeodominio/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos mdx , Músculo Esquelético/metabolismo , Músculo Esquelético/patología , Músculo Esquelético/efectos de la radiación , Distrofia Muscular Animal/metabolismo , Músculos Oculomotores/metabolismo , Factor de Transcripción PAX7/metabolismo , Fenotipo , Células Satélite del Músculo Esquelético/metabolismo , Células Satélite del Músculo Esquelético/patología , Células Satélite del Músculo Esquelético/efectos de la radiación , Células Madre/metabolismo , Células Madre/efectos de la radiación , Factores de Transcripción/metabolismo , Proteína del Homeodomínio PITX2RESUMEN
Previous studies showed that monochromatic green light stimuli during embryogenesis accelerated posthatch body weight (BW) and pectoral muscle growth of broilers. In this experiment, we further investigated the morphological and molecular basis of this phenomenon. Fertile broiler eggs (Arbor Acres, n=880) were pre-weighed and randomly assigned to 1 of the 2 incubation treatment groups: (1) dark condition (control group), and (2) monochromatic green light group (560 nm). The monochromatic lighting systems sourced from light-emitting diode lamps and were equalized at the intensity of 15 lx at eggshell level. The dark condition was set as a commercial control from day 1 until hatching. After hatch, 120 male 1-day-old chicks from each group were housed under incandescent white light with an intensity of 30 lx at bird-head level. No effects of light stimuli during embryogenesis on hatching time, hatchability, hatching weight and bird mortality during the feeding trial period were observed in the present study. Compared with the dark condition, the BW, pectoral muscle weight and myofiber cross-sectional areas were significantly greater on 7-day-old chicks incubated under green light. Green light also increased the satellite cell mitotic activity of pectoral muscle on 1- and 3-day-old birds. In addition, green light upregulated MyoD, myogenin and myostatin mRNA expression in late embryos and/ or newly hatched chicks. These data suggest that stimulation with monochromatic green light during incubation promote muscle growth by enhancing proliferation and differentiation of satellite cells in late embryonic and newly hatched stages. Higher expression of myostatin may ultimately help prevent excessive proliferation and differentiation of satellite cells in birds incubated under green light.
Asunto(s)
Embrión de Pollo/efectos de la radiación , Pollos/fisiología , Regulación del Desarrollo de la Expresión Génica/efectos de la radiación , Luz , Mitosis/efectos de la radiación , Músculo Esquelético/crecimiento & desarrollo , Células Satélite del Músculo Esquelético/citología , Animales , Proliferación Celular/efectos de la radiación , Embrión de Pollo/embriología , Inmunohistoquímica/veterinaria , Incubadoras/veterinaria , Masculino , Músculo Esquelético/efectos de la radiación , Proteína MioD/metabolismo , Miogenina/metabolismo , Miostatina/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Células Satélite del Músculo Esquelético/efectos de la radiaciónRESUMEN
Skeletal muscles are commonly exposed to radiation for diagnostic procedures and the treatment of cancers and heterotopic bone formation. Few studies have considered the impact of clinical doses of radiation on the ability of satellite cells (myogenic stem cells) to proliferate, differentiate and contribute to recovering/maintaining muscle mass. The primary objective of this study was to determine whether the proliferation of irradiated satellite cells could be rescued by manipulating NO levels via pharmacological approaches and mechanical stretch (which is known to increase NO levels). We used both SNP (NO donor) and PTIO (NO scavenger) to manipulate NO levels in satellite cells. We observed that SNP was highly effective in rescuing the proliferation of irradiated satellite cells, especially at doses less than 5 Gy. The potential importance of NO was further illustrated by the effects of PTIO, which completely inhibited the rescue effect of SNP. Mechanical cyclic stretch was found to produce significant increases in NO levels of irradiated satellite cells, and this was associated with a robust increase in satellite cell proliferation. The effects of both radiation and NO on two key myogenic regulatory factors (MyoD and myogenin) were also explored. Irradiation of satellite cells produced a significant increase in both MyoD and myogenin, effects that were mitigated by manipulating NO levels via SNP. Given the central role of myogenic regulatory factors in the proliferation and differentiation of satellite cells, the findings of the current study underscore the need to more fully understand the relationship between radiation, NO and the functionality of satellite cells.
Asunto(s)
Óxido Nítrico/metabolismo , Células Satélite del Músculo Esquelético/metabolismo , Células Satélite del Músculo Esquelético/efectos de la radiación , Animales , Fenómenos Biomecánicos , Recuento de Células , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/efectos de la radiación , Proliferación Celular/efectos de los fármacos , Proliferación Celular/efectos de la radiación , Óxidos N-Cíclicos/farmacología , Relación Dosis-Respuesta en la Radiación , Depuradores de Radicales Libres/farmacología , Rayos gamma , Imidazoles/farmacología , Masculino , Factores Reguladores Miogénicos/metabolismo , Óxido Nítrico/biosíntesis , Nitroprusiato/farmacología , Ratas , Ratas Sprague-Dawley , Células Satélite del Músculo Esquelético/citología , Células Satélite del Músculo Esquelético/efectos de los fármacosRESUMEN
Skeletal muscles are the organ of movement, and their growth, regeneration and maintenance are dependent in large part on a population of myogenic stem cells known as satellite cells. Skeletal muscles and these resident myogenic stem cells (i.e., satellite cells) are commonly exposed to significant doses of radiation during diagnostic procedures and/or during the radiotherapeutic management of cancer. The main objective of this study was to examine the effects of clinically relevant doses of γ radiation on satellite cell survival and proliferation, cell cycle regulation, apoptosis, DNA double-strand break repair, oxidative stress and NO production. Overall, our findings demonstrate that doses of γ radiation ≥5 Gy reduced satellite cell numbers by at least 70% due in part to elevated apoptosis and the inhibition of cell cycle progression. Radiation was also found to cause a significant and persistent increase in the level of reactive oxygen and nitrogen species. Interestingly, and within this backdrop of elevated oxidative stress, similar doses were found to produce substantial reductions in the levels of nitric oxide (NO). Proliferation of satellite cells has been shown to depend in part on the production of NO, and our findings give rise to the possibility that radiation-induced reductions in NO levels may provide a mechanism for the inhibition of satellite cell proliferation in vitro and possibly the regrowth of skeletal muscle exposed during clinical irradiation procedures.
Asunto(s)
Apoptosis/efectos de la radiación , Ciclo Celular/efectos de la radiación , Estrés Oxidativo/efectos de la radiación , Células Satélite del Músculo Esquelético/citología , Células Satélite del Músculo Esquelético/efectos de la radiación , Animales , Proliferación Celular/efectos de la radiación , Supervivencia Celular/efectos de la radiación , Roturas del ADN de Doble Cadena/efectos de la radiación , Relación Dosis-Respuesta en la Radiación , Femenino , Rayos gamma , Histonas/metabolismo , Cinética , Óxido Nítrico/biosíntesis , Ratas , Células Satélite del Músculo Esquelético/metabolismo , Transducción de Señal/efectos de la radiaciónRESUMEN
Low-level laser (LLL) irradiation promotes proliferation of muscle satellite cells, angiogenesis and expression of growth factors. Satellite cells, angiogenesis and growth factors play important roles in the regeneration of muscle. The objective of this study was to examine the effect of LLL irradiation on rat gastrocnemius muscle recovering from disuse muscle atrophy. Eight-week-old rats were subjected to hindlimb suspension for 2 weeks, after which they were released and recovered. During the recovery period, rats underwent daily LLL irradiation (Ga-Al-As laser; 830 nm; 60 mW; total, 180 s) to the right gastrocnemius muscle through the skin. The untreated left gastrocnemius muscle served as the control. In conjunction with LLL irradiation, 5-bromo-2-deoxyuridine (BrdU) was injected subcutaneously to label the nuclei of proliferating cells. After 2 weeks, myofibre diameters of irradiated muscle increased in comparison with those of untreated muscle, but did not recover back to normal levels. Additionally, in the superficial region of the irradiated muscle, the number of capillaries and fibroblast growth factor levels exhibited significant elevation relative to those of untreated muscle. In the deep region of irradiated muscle, BrdU-positive nuclei of satellite cells and/or myofibres increased significantly relative to those of the untreated muscle. The results of this study suggest that LLL irradiation can promote recovery from disuse muscle atrophy in association with proliferation of satellite cells and angiogenesis.
Asunto(s)
Terapia por Luz de Baja Intensidad , Músculo Esquelético/patología , Músculo Esquelético/efectos de la radiación , Atrofia Muscular/radioterapia , Animales , Bromodesoxiuridina/metabolismo , Capilares/patología , Capilares/efectos de la radiación , Proliferación Celular/efectos de la radiación , Factor 2 de Crecimiento de Fibroblastos/metabolismo , Suspensión Trasera , Factor I del Crecimiento Similar a la Insulina/metabolismo , Masculino , Músculo Esquelético/irrigación sanguínea , Músculo Esquelético/metabolismo , Atrofia Muscular/etiología , Atrofia Muscular/metabolismo , Atrofia Muscular/patología , Mioblastos Esqueléticos/metabolismo , Mioblastos Esqueléticos/patología , Mioblastos Esqueléticos/efectos de la radiación , Miofibrillas/patología , Neovascularización Fisiológica/efectos de la radiación , Ratas , Ratas Wistar , Células Satélite del Músculo Esquelético/metabolismo , Células Satélite del Músculo Esquelético/patología , Células Satélite del Músculo Esquelético/efectos de la radiaciónRESUMEN
The purpose of this time-course study was to determine whether satellite cell ablation within rat tibialis anterior (TA) muscles exposed to short-term chronic low-frequency stimulation (CLFS) would limit fast-to-slow fibre type transformations. Satellite cells of the left TA were ablated by exposure to gamma-irradiation before 1, 2, 5 or 10 days of CLFS and 1 week later where required. Control groups received only CLFS or a sham operation. Continuous infusion of 5-bromo-2'-deoxyuridine revealed that CLFS first induced an increase in satellite cell proliferation at 1 day, up to a maximum at 10 days over control (mean +/- SEM, 5.7 +/- 0.7 and 20.4 +/- 1.0 versus 1.5 +/- 0.2 mm(-2), respectively, P < 0.007) that was abolished by gamma-irradiation. Myosin heavy chain mRNA, immunohistochemical and sodium dodecyl sulfate polyacrylamide gel electrophoresis analyses revealed CLFS-induced fast-to-slow fibre type transformation began at 5 days and continued at 10 days; in those muscles that were also exposed to gamma-irradiation, attenuation occurred within the fast fibre population, and the final fast-twitch to slow-twitch adaptation did not occur. These findings indicate satellite cells play active and obligatory roles early on in the time course during skeletal muscle fibre type adaptations to CLFS.
Asunto(s)
Fibras Musculares de Contracción Rápida/fisiología , Fibras Musculares de Contracción Lenta/fisiología , Células Satélite del Músculo Esquelético/fisiología , Adaptación Fisiológica , Animales , Proliferación Celular/efectos de la radiación , Estimulación Eléctrica , Rayos gamma , Antígenos de Histocompatibilidad/metabolismo , Masculino , Fibras Musculares de Contracción Rápida/ultraestructura , Fibras Musculares de Contracción Lenta/ultraestructura , Músculo Esquelético/metabolismo , Isoformas de Proteínas/metabolismo , Ratas , Ratas Wistar , Células Satélite del Músculo Esquelético/efectos de la radiaciónRESUMEN
Skeletal muscle work hypertrophy is usually connected with muscle progenitor SC (satellite cells) activation with subsequent incorporation their nuclei into myofibers. Passive stretch of unloaded muscle was earlier established to prevent atrophic processes and be accompanied by enhanced protein synthesis. We hypothesized that elimination of SC proliferation capacity by gamma-irradiation would partly preavent stretched muscle fiber capability to maintain their size under condition of gravitational unloading. To assess the role of muscle progenitor (satellite) cells in development of passive stretch preventive effect SC proliferation was suppressed by local exposure to ionizing radiation (2500 Rad) and then subsequent hindlimb suspension or hindlimb suspension with concomitant passive stretch were carried out. Reduction of myofiber cross-sectional area and decrease in myo-nuclei number accompanying unloaded muscle atrophy were completely abolished by passive stretch both in irradiated and sham-treated animals. We concluded that satellite cells did not make essential contribution to passive stretch preventive action under condition of simulated weightlessness.