The isolation of syaptic myonuclei and Schwann cell nuclei for DNA analysis from human skeletal muscle

The neuromuscular junction is a critical component of the motor system that enables transmission of impulses from the alpha-motor nerve to muscle fibers. Cerebral palsy is a neuromotor disorder that has been characterized by deficits in the central nervous system and, more recently, disruptions in the peripheral nervous system at the neuromuscular junction. The specific mechanisms accounting for neuromuscular junction disruption in cerebral palsy are not known, but the development and maintenance of the neuromuscular junctions is a complex process involving alterations in gene expression of synaptic myonuclei and terminal Schwann cells and the development of a highly-regulated synaptic structure. To understand neuromuscular junction abnormalities associated with cerebral palsy, a method for assessing the regulation of gene expression in synapse-associated nuclei is needed. This work focuses on development of a technique to isolate synaptic myonuclei and terminal Schwann cell nuclei from human skeletal muscle biopsies to enable epigenetic analyses. Using tissue dissociation and immunological separation techniques, a method to isolate synaptic myonuclei and terminal Schwann cells is accomplished using fluorescence-activated and magnetic-activated sorting technologies. Nuclear material separated from muscles of patients with different clinical diagnoses using this technique may provide a critical approach to studying the epigenetic regulation of neuromuscular junction development and maintenance in both health and disease.

Grantor dc:publisher

University of Delaware

Year dc:date.issued

2015

Author dc:creator

• Barnes, Daniel T.

DOI

https://doi.org/10.58088/s66n-c857

OAI identifier oai:identifier

oai:udspace.udel.edu:19716/17654