RESUMEN
Whole-body knock-out of Cu,Zn superoxide dismutase (Sod1KO) results in accelerated, age-related loss of muscle mass and function associated with neuromuscular junction (NMJ) breakdown similar to sarcopenia. In order to determine whether altered redox in motor neurons underlies this phenotype, an inducible neuron-specific deletion of Sod1 (i-mnSod1KO) was compared with wild-type (WT) mice of different ages (adult, mid-age, and old) and whole-body Sod1KO mice. Nerve oxidative damage, motor neuron numbers and structural changes to neurons and NMJ were examined. Tamoxifen-induced deletion of neuronal Sod1 from two months of age. No specific effect of a lack of neuronal Sod1 was seen on markers of nerve oxidation (electron paramagnetic resonance of an in vivo spin probe, protein carbonyl, or protein 3-nitrotyrosine contents). i-mnSod1KO mice showed increased denervated NMJ, reduced numbers of large axons and increased number of small axons compared with old WT mice. A large proportion of the innervated NMJs in old i-mnSod1KO mice displayed a simpler structure than that seen in adult or old WT mice. Thus, previous work showed that neuronal deletion of Sod1 induced exaggerated loss of muscle in old mice, and we report that this deletion leads to a specific nerve phenotype including reduced axonal area, increased proportion of denervated NMJ, and reduced acetyl choline receptor complexity. Other changes in nerve and NMJ structure seen in the old i-mnSod1KO mice reflect aging of the mice.
Asunto(s)
Músculo Esquelético , Unión Neuromuscular , Ratones , Animales , Músculo Esquelético/fisiología , Unión Neuromuscular/metabolismo , Neuronas Motoras/metabolismo , Superóxido Dismutasa-1/genética , Superóxido Dismutasa-1/metabolismo , Axones/metabolismo , Ratones Transgénicos , Superóxido Dismutasa/genéticaRESUMEN
Motor unit remodelling involving repeated denervation and re-innervation occurs throughout life. The efficiency of this process declines with age contributing to neuromuscular deficits. This study investigated differentially expressed genes (DEG) in muscle following peroneal nerve crush to model motor unit remodelling in C57BL/6 J mice. Muscle RNA was isolated at 3 days post-crush, RNA libraries were generated using poly-A selection, sequenced and analysed using gene ontology and pathway tools. Three hundred thirty-four DEG were found in quiescent muscle from (26mnth) old compared with (4-6mnth) adult mice and these same DEG were present in muscle from adult mice following nerve crush. Peroneal crush induced 7133 DEG in muscles of adult and 699 DEG in muscles from old mice, although only one DEG (ZCCHC17) was found when directly comparing nerve-crushed muscles from old and adult mice. This analysis revealed key differences in muscle responses which may underlie the diminished ability of old mice to repair following nerve injury.
Asunto(s)
Lesiones por Aplastamiento , Desnervación Muscular , Envejecimiento/genética , Animales , Ratones , Ratones Endogámicos C57BL , Músculo Esquelético/inervación , Compresión Nerviosa , Regeneración Nerviosa/fisiología , ARN , TranscriptomaAsunto(s)
Poliangitis Microscópica/diagnóstico , Paniculitis/patología , Piel/patología , Diagnóstico Diferencial , Femenino , Humanos , Inmunosupresores/uso terapéutico , Poliangitis Microscópica/complicaciones , Poliangitis Microscópica/tratamiento farmacológico , Poliangitis Microscópica/patología , Persona de Mediana Edad , Paniculitis/tratamiento farmacológico , Paniculitis/etiología , Insuficiencia Renal/etiologíaRESUMEN
To determine the role of denervation and motor unit turnover in the age-related increase in skeletal muscle oxidative stress, the hydrogen peroxide (H2O2) specific, genetically-encoded, fluorescent cyto-HyPer2 probe was expressed in mouse anterior tibialis (AT) muscle and compared with ex vivo measurements of mitochondrial oxidant generation. Crush of the peroneal nerve induced increased mitochondrial peroxide generation, measured in permeabilised AT fibers ex vivo and intra vital confocal microscopy of cyto-HyPer2 fluorescence showed increased cytosolic H2O2 in a sub-set (~24%) of individual fibers associated with onset of fiber atrophy. In comparison, mitochondrial peroxide generation was also increased in resting muscle from old (26 month) mice compared with adult (6-8 month) mice, but no age effect on fiber cytosolic H2O2 in vivo was seen. Thus ageing is associated with an increased ability of muscle fibers to maintain cytosolic redox homeostasis in the presence of denervation-induced increase in mitochondrial peroxide generation.
Asunto(s)
Envejecimiento/metabolismo , Peróxido de Hidrógeno/metabolismo , Sondas Moleculares/metabolismo , Fibras Musculares Esqueléticas/metabolismo , Animales , Colorantes Fluorescentes , Masculino , Ratones , Mitocondrias/metabolismo , Atrofia Muscular/metabolismo , Compresión Nerviosa , Unión Neuromuscular/metabolismo , Oxidantes/metabolismo , Estrés Oxidativo , Sarcopenia/metabolismoRESUMEN
A technique for the amputation and immediate reimplantation of rats legs was studied using a microsurgical technique. Complete survival of the reimplanted leg was achieved by basing the blood supply on anastomoses of arteries about 0.4 mm in external diameter. Anastomoses of vessels in that range by interrupted sutures of 10/0 and 11/0 black monofilament nylon proved to be compatible with leg survival. The ultimate survival of the reimplanted limbs depended upon microsurgical technique, preoperative and operative heparinization and external protection during the recovery phase. Animals are alive and using the replanted limb, several months postoperatively.