Glucose metabolism in rats submitted to skeletal muscle denervation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Muscle reinnervation in one or two stages?: experimental study in rats with end-to-side nerve graft

OBJETIVO: Comparar a reinervação muscular com enxerto de nervo em um e dois tempos operatórios, utilizando a neurorrafia término-lateral (NTL) sem lesão do nervo doador. MÉTODOS: Vinte ratos foram distribuídos em quatro grupos. O grupo 1 (G1), um estágio, recebeu o enxerto que foi suturado ao nervo tibial (NT), por meio de NTL, e seu coto livre foi suturado por NTL ao coto distal do nervo peroneal (NP), seccionado a um centímetro do NT, na mesma cirurgia. O grupo 2 (G2), dois estágios, recebeu o enxerto de nervo na primeira cirurgia, como já descrito. Dois meses depois, na segunda cirurgia, o NP foi seccionado e seu coto distal ligado ao coto distal do enxerto como em G1. O grupo controle de normalidade (Gn) recebeu o enxerto da mesma forma, apenas. E o grupo controle de denervação (Gd), além de receber o enxerto, teve o NP seccionado e seus cotos sepultados na musculatura adjacente, com a finalidade de denervar o músculo tibial cranial (MTC), alvo deste estudo. Os parâmetros utilizados para avaliar a reinervação do MTC foram massa muscular, diâmetro mínimo da fibra muscular e área. RESULTADOS: O grupo G2 apresentou superioridade (p<0,0001) em relação ao G1 na massa do MTC, no diâmetro mínimo e na área das fibras musculares. Na comparação entre os quatro grupos, estes mesmos parâmetros tiveram sua expressão máxima em Gn e mínima em Gd, como era esperado. CONCLUSÃO: A reinervação muscular em dois estágios apresenta melhor resultado quando comparada à técnica em um tempo.

Effect of electrical stimulation of the cranial tibial muscle after end-to-side neurorrhaphy of the peroneal nerve in rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Morphometric evaluation of the rat testis, epididymis and vas deferens following chemical sympathectomy with guanethidine

Selective chemical sympathectomy of the internal sex organs of adult male rats was undertaken by long term administration of low doses of guanethidine. The spermatogenic activity of the testis was unaffected by treatment. Examination of the vas deferens using morphometric methods revealed a marked increase in luminal area in contrast to a decrease in muscle layer area and in epithelial height. This is morphological evidence of sperm accumulation caused by a disorder in ductal contractile activity. No structural changes were observed in the epididymis. However, the concentration of spermatozoa in the sperm suspension stored in the cauda epididymidis was significantly increased in denervated rats. This result is discussed in terms of a sympathetic control of resorption mechanisms in the epididymis.

Marinesco-Sjögren syndrome protein SIL1 regulates motor neuron subtype-selective ER stress in ALS.

Mechanisms underlying motor neuron subtype-selective endoplasmic reticulum (ER) stress and associated axonal pathology in amyotrophic lateral sclerosis (ALS) remain unclear. Here we show that the molecular environment of the ER between motor neuron subtypes is distinct, with characteristic signatures. We identify cochaperone SIL1, mutated in Marinesco-Sjögren syndrome (MSS), as being robustly expressed in disease-resistant slow motor neurons but not in ER stress-prone fast-fatigable motor neurons. In a mouse model of MSS, we demonstrate impaired ER homeostasis in motor neurons in response to loss of SIL1 function. Loss of a single functional Sil1 allele in an ALS mouse model (SOD1-G93A) enhanced ER stress and exacerbated ALS pathology. In SOD1-G93A mice, SIL1 levels were progressively and selectively reduced in vulnerable fast-fatigable motor neurons. Mechanistically, reduction in SIL1 levels was associated with lowered excitability of fast-fatigable motor neurons, further influencing expression of specific ER chaperones. Adeno-associated virus-mediated delivery of SIL1 to familial ALS motor neurons restored ER homeostasis, delayed muscle denervation and prolonged survival.

A mitochondrial perspective on striated muscle physiopathology: insights from sepsis, denervation, and dystrophinopathies.

La mitochondrie est de plus en plus reconnue pour sa contribution à la dégénerescence musculaire. Les dysfonctions mitochondriales, en plus de causer une défaillance énergétique, contribuent à la signalisation apoptotique, stimule la production de ROS et peuvent induire une surcharge calcique. Ces caractéristiques sont tous reliées à certains types de myopathies. Cette thèse met en lumières comment certaines dysfonctions mitochondriales peuvent intervenir dans la pathogenèse de diverses myopathies. Nous démontrons que les dysfonctions mitochondriales sont impliqués dans l’atrophie dû à la perte d’innervation. Par contre, la désensabilisation de l’ouverture du pore mitochondrial de transition de perméabilité, via ablation génétique de cyclophiline-D, ne prévient ni la signalisation apoptotique mitochondrial ni l’atrophie. Nous avons aussi observé des dysfonctions mitochondriales dans le muscle atteint de dystrophie musculaire de Duchenne qui furent améliorés suite à une transfection de PGC1-α, laquelle résulta aussi en une amélioration de la pathologie. Finalement, nous démontrons que le recyclage de mitochondrie par les voies de mitophagies et de contrôles de la qualité impliquant Parkin et possiblement d’autres voies de signalisation inconnues sont cruciales au recouvrement cardiaqe lors d’un choc septique.

Muscle-specific mutations accumulate with aging in critical human mtDNA control sites for replication.

The recently discovered aging-dependent large accumulation of point mutations in the human fibroblast mtDNA control region raised the question of their occurrence in postmitotic tissues. In the present work, analysis of biopsied or autopsied human skeletal muscle revealed the absence or only minimal presence of those mutations. By contrast, surprisingly, most of 26 individuals 53 to 92 years old, without a known history of neuromuscular disease, exhibited at mtDNA replication control sites in muscle an accumulation of two new point mutations, i.e., A189G and T408A, which were absent or marginally present in 19 individuals younger than 34 years. These two mutations were not found in fibroblasts from 22 subjects 64 to 101 years of age (T408A), or were present only in three subjects in very low amounts (A189G). Furthermore, in several older individuals exhibiting an accumulation in muscle of one or both of these mutations, they were nearly absent in other tissues, whereas the most frequent fibroblast-specific mutation (T414G) was present in skin, but not in muscle. Among eight additional individuals exhibiting partial denervation of their biopsied muscle, four subjects >80 years old had accumulated the two muscle-specific point mutations, which were, conversely, present at only very low levels in four subjects <or =40 years old. The striking tissue specificity of the muscle mtDNA mutations detected here and their mapping at critical sites for mtDNA replication strongly point to the involvement of a specific mutagenic machinery and to the functional relevance of these mutations.

MuRF1 is a muscle fiber-type II associated factor and together with MuRF2 regulates type-II fiber trophicity and maintenance

MuRF1 is a member of the RBCC (RING, B-box, coiled-coil) superfamily that has been proposed to act as an atrogin during muscle wasting. Here, we show that MuRF1 is preferentially induced in type-II muscle fibers after denervation. Fourteen days after denervation, MuRF1 protein was further elevated but remained preferentially expressed in type-II muscle fibers. Consistent with a fiber-type dependent function of MuRF1, the tibialis anterior muscle (rich in type-II muscle fibers) was considerably more protected in MuRF1-KO mice from muscle wasting when compared to soleus muscle with mixed fiber-types. We also determined fiber-type distributions in MuRF1/MuRF2 double-deficient KO (dKO) mice, because MuRF2 is a close homolog of MuRF1. MuRF1/MuRF2 dKO mice showed a profound loss of type-II fibers in soleus muscle. As a potential mechanism we identified the interaction of MuRF1/MuRF2 with myozenin-1, a calcineurin/NFAT regulator and a factor required for maintenance of type-II muscle fibers. MuRF1/MuRF2 dKO mice had lost myozenin-1 expression in tibialis anterior muscle, implicating MuRF1/MuRF2 as regulators of the calcineurin/NFAT pathway. In summary, our data suggest that expression of MuRF1 is required for remodeling of type-II fibers under pathophysiological stress states, whereas MuRF1 and MuRF2 together are required for maintenance of type-II fibers, possibly via the regulation of myozenin-1. (C) 2010 Elsevier Inc. All rights reserved.

Morphologic alterations resulting from denervation of the diaphragm in rats

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Myenteric Denervation Downregulates Galectin-1 and-3 Expression in Gastric Carcinogenesis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Morphological aspects of rabbit masseter muscle after cervical sympathectomy

The objective of this paper was to study the effect of sympathetic innervation on morphological and histochemical aspects of skeletal muscle tissue. Rabbit masseter muscle was studied using histochemical and immunohistochemical methods for periods of up to 18 months post-sympathectomy. The morphological and enzymatic characteristics of control masseter muscles were similar on both the left and right sides. The main features were muscle fibres with a mosaic pattern and a predominance of type IIa fibres, followed by type I. Type IIb fibres showed very low frequency. Sympathectomized animals showed varying degrees of metabolic and morphological alterations, especially 18 months after sympathectomy. The first five groups showed a higher frequency of type I fibres, whilst the oldest group showed a higher frequency of type lib fibres. In the oldest group, a significant variation in fibre diameter was observed. Many fibres showed small diameter, atrophy, hypertrophy, splitting, and necrosis. Areas with fibrosis were observed. Thus cervical sympathectomy induced morphological alterations in the masseter muscles. These alterations were, in part, similar to both denervation and myopathy. These findings indicate that sympathetic innervation contributes to the maintenance of the morphological and metabolic features of masseter muscle fibres.

Morphological aspects of rabbit masseter muscle after cervical sympathectomy

The objective of this paper was to study the effect of sympathetic innervation on morphological and histochemical aspects of skeletal muscle tissue. Rabbit masseter muscle was studied using histochemical and immunohistochemical methods for periods of up to 18 months post-sympathectomy. The morphological and enzymatic characteristics of control masseter muscles were similar on both the left and right sides. The main features were muscle fibres with a mosaic pattern and a predominance of type IIa fibres, followed by type I. Type IIb fibres showed very low frequency. Sympathectomized animals showed varying degrees of metabolic and morphological alterations, especially 18 months after sympathectomy. The first five groups showed a higher frequency of type I fibres, whilst the oldest group showed a higher frequency of type IIb fibres. In the oldest group, a significant variation in fibre diameter was observed. Many fibres showed small diameter, atrophy, hypertrophy, splitting, and necrosis. Areas with fibrosis were observed. Thus cervical sympathectomy induced morphological alterations in the masseter muscles. These alterations were, in part, similar to both denervation and myopathy. These findings indicate that sympathetic innervation contributes to the maintenance of the morphological and metabolic features of masseter muscle fibres.

Left cardiac sympathetic denervation for treatment of symptomatic systolic heart failure patients: a pilot study

To evaluate the feasibility, safety, and potential beneficial effects of left cardiac sympathetic denervation (LCSD) in systolic heart failure (HF) patients. In this prospective, randomized pilot study, inclusion criteria were New York Heart Association (NYHA) functional class II or III, left ventricular ejection fraction (LVEF) 40, sinus rhythm, and resting heart rate 65 b.p.m., despite optimal medical therapy (MT). Fifteen patients were randomly assigned either to MT alone or MT plus LCSD. The primary endpoint was safety, measured by mortality in the first month of follow-up and morbidity according to pre-specified criteria. Secondary endpoints were exercise capacity, quality of life, LVEF, muscle sympathetic nerve activity (MSNA), brain natriuretic peptide (BNP) levels and 24 h Holter mean heart rate before and after 6 months. We studied clinical effects in long-term follow-up. Ten patients underwent LCSD. There were no adverse events attributable to surgery. In the LCSD group, LVEF improved from 25 6.6 to 33 5.2 (P 0.03); 6 min walking distance improved from 167 35 to 198 47 m (P 0.02). Minnesota Living with Heart Failure Questionnaire (MLWHFQ) score physical dimension changed from 21 5 to 15 7 (P 0.06). The remaining analysed variables were unchanged. During 848 549 days of follow-up, in the MT group, three patients either died or underwent cardiac transplantation (CT), while in the LCSD group six were alive without CT. LCSD was feasible and seemed to be safe in systolic HF patients. Its beneficial effects warrant the development of a larger randomized trial. Trail registration: NCT01224899.

Muscle-specific mutations accumulate with aging in critical human mtDNA control sites for replication

The recently discovered aging-dependent large accumulation of point mutations in the human fibroblast mtDNA control region raised the question of their occurrence in postmitotic tissues. In the present work, analysis of biopsied or autopsied human skeletal muscle revealed the absence or only minimal presence of those mutations. By contrast, surprisingly, most of 26 individuals 53 to 92 years old, without a known history of neuromuscular disease, exhibited at mtDNA replication control sites in muscle an accumulation of two new point mutations, i.e., A189G and T408A, which were absent or marginally present in 19 individuals younger than 34 years. These two mutations were not found in fibroblasts from 22 subjects 64 to 101 years of age (T408A), or were present only in three subjects in very low amounts (A189G). Furthermore, in several older individuals exhibiting an accumulation in muscle of one or both of these mutations, they were nearly absent in other tissues, whereas the most frequent fibroblast-specific mutation (T414G) was present in skin, but not in muscle. Among eight additional individuals exhibiting partial denervation of their biopsied muscle, four subjects >80 years old had accumulated the two muscle-specific point mutations, which were, conversely, present at only very low levels in four subjects ≤40 years old. The striking tissue specificity of the muscle mtDNA mutations detected here and their mapping at critical sites for mtDNA replication strongly point to the involvement of a specific mutagenic machinery and to the functional relevance of these mutations.