Dataset Information

Endomicroscopy and electromyography of neuromuscular junctions in situ.

ABSTRACT:

Objective

Electromyography (EMG) is used routinely to diagnose neuromuscular dysfunction in a wide range of peripheral neuropathies, myopathies, and neuromuscular degenerative diseases including motor neuron diseases such as amyotrophic lateral sclerosis (ALS). Definitive neurological diagnosis may also be indicated by the analysis of pathological neuromuscular innervation in motor-point biopsies. Our objective in this study was to preempt motor-point biopsy by combining live imaging with electrophysiological analysis of slow degeneration of neuromuscular junctions (NMJs) in vivo.

Methods

We combined conventional needle electromyography with fiber-optic confocal endomicroscopy (CEM), using an integrated hand-held, 1.5-mm-diameter probe. We utilized as a test bed, various axotomized muscles in the hind limbs of anaesthetized, double-homozygous thy1.2YFP16: Wld (S) mice, which coexpress the Wallerian-degeneration Slow (Wld(S)) protein and yellow fluorescent protein (YFP) in motor neurons. We also tested exogenous vital stains, including Alexa488-?-bungarotoxin; the styryl pyridinium dye 4-Di-2-Asp; and a GFP conjugate of botulinum toxin Type A heavy chain (GFP-HcBoNT/A).

Results

We show that an integrated EMG/CEM probe is effective in longitudinal evaluation of functional and morphological changes that take place over a 7-day period during axotomy-induced, slow neuromuscular synaptic degeneration. EMG amplitude declined in parallel with overt degeneration of motor nerve terminals. EMG/CEM was safe and effective when nerve terminals and motor endplates were selectively stained with vital dyes.

Interpretation

Our findings constitute proof-of-concept, based on live imaging in an animal model, that combining EMG/CEM may be useful as a minimally invasive precursor or alternative to motor-point biopsy in neurological diagnosis and for monitoring local administration of potential therapeutics.

SUBMITTER: Brown R

PROVIDER: S-EPMC4265058 | biostudies-literature | 2014 Nov

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Endomicroscopy and electromyography of neuromuscular junctions in situ.

Brown Rosalind R Dissanayake Kosala N KN Skehel Paul A PA Ribchester Richard R RR

Annals of clinical and translational neurology 20141010 11

<h4>Objective</h4>Electromyography (EMG) is used routinely to diagnose neuromuscular dysfunction in a wide range of peripheral neuropathies, myopathies, and neuromuscular degenerative diseases including motor neuron diseases such as amyotrophic lateral sclerosis (ALS). Definitive neurological diagnosis may also be indicated by the analysis of pathological neuromuscular innervation in motor-point biopsies. Our objective in this study was to preempt motor-point biopsy by combining live imaging wit ...[more]

PMID: 25540801

Similar Datasets

Project description:BackgroundThere is currently no objective and accurate clinical assessment of reticular neuromuscular control in healthy subjects or patients with upper motor neuron injury. As a result, clinical dysfunctions of neuromuscular control could just be semi-quantified, efficacies and mechanisms of various therapies for neuromuscular control improving are difficult to verify.MethodsFourteen healthy participants were required to maintain standing balance in the kinetostatics model of Gusu Constraint Standing Training (GCST). A backward and upward constraint force was applied to their trunk at 0°, 20° and 25°, respectively. The multiplex recurrence network (MRN) was applied to analyze the surface electromyography signals of 16 muscles of bilateral lower limbs during the tests. Different levels of MRN network indices were utilized to assess reticular neuromuscular control.ResultsCompared with the 0° test, the MRN indices related to muscle coordination of bilateral lower limbs, of unilateral lower limb and of inter limbs showed significant increase when participants stood in 20° and 25° tests (P < 0.05). The indices related to muscle contribution of gluteal, anterior thigh and calf muscles significantly increased when participants stood in 20° and 25° tests (P < 0.05).ConclusionsThis study applied the dynamical network indices of MRN to analyze the changes of neuromuscular control of lower limbs of healthy participants in the kinetostatics model of GCST. Results showed that the overall coordination of lower limb muscles would be significantly enhanced during performing GCST, partly by the enhancement of neuromuscular control of single lower limb, and partly by the enhancement of joint control across lower limbs. In particular, the muscles in buttocks, anterior thighs and calves played a more important role in the overall coordination, and their involvement was significantly increased. The MRN could provide details of control at the bilateral lower limbs, unilateral lower limb, inter limbs, and single muscle levels, and has the potential to be a new tool for assessing the reticular neuromuscular control. Trial registration ChiCTR2100055090.

Project description:Desmin, the major intermediate filament (IF) protein in muscle cells, interlinks neighboring myofibrils and connects the whole myofibrillar apparatus to myonuclei, mitochondria, and the sarcolemma. However, desmin is also known to be enriched at postsynaptic membranes of neuromuscular junctions (NMJs). The pivotal role of the desmin IF cytoskeletal network is underscored by the fact that over 100 mutations of the human DES gene cause hereditary and sporadic myopathies and cardiomyopathies. A subgroup of human desminopathies comprises autosomal recessive cases resulting in complete abolition of desmin protein. In these patients, who display a more severe phenotype than the autosomal dominant cases, it has been reported that some individuals also suffer from a myasthenic syndrome in addition to the classical occurrence of myopathy and cardiomyopathy. Since further studies on the NMJ pathology is hampered by the lack of available human striated muscle biopsy specimens, we exploited homozygous desmin knock-out mice which closely mirror the striated muscle pathology of human patients lacking desmin protein. Here, we report on the impact of the lack of desmin on the structure and function of NMJs and on the transcription of genes coding for postsynaptic proteins. Desmin knock-out mice display a fragmentation of NMJs in soleus, but not in extensor digitorum longus muscle. Moreover, soleus muscle fibers show larger NMJs. Further, transcription levels of acetylcholine receptor (AChR) genes are increased in muscles from desmin knock-out mice, especially of the AChR subunit, which is known as a marker of muscle fiber regeneration. Electrophysiological recordings depicted a pathological decrement of nerve-dependent endplate potentials and a faster rise time of the nerve-independent miniature endplate potentials. The latter is indicating an enhanced opening time of the AChR channels. Our study highlights the essential role of desmin for the structural and functional integrity of mammalian neuromuscular junctions.

Dataset Information

Endomicroscopy and electromyography of neuromuscular junctions in situ.

Objective

Methods

Results

Interpretation

Publications

Endomicroscopy and electromyography of neuromuscular junctions in situ.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets