title: Compartmental microfluidic system for studying muscle-neuron communication and neuromuscular junction maintenance.
abstract: Molecular communication between the motoneuron and the muscle is vital for neuromuscular junction (NMJ) formation and maintenance. Disruption in the structure and function of NMJs is a hallmark of various neurodegenerative processes during both development and pathological events. Still due to the complexity of this process, it is very difficult to elucidate the cellular mechanisms underlying it, generating a keen interest for developing better tools for investigating it. Here we describe a simplified method to study mechanisms of NMJs formation, maintenance and disruption. A spinal cord explant from mice expressing the Hb9::GFP motoneuron marker is plated on one side of a compartmental chamber, and myotubes derived from muscle satellite progenitor cells are plated on the other. The GFP labeled motoneurons extend their axons via microgrooves in the chamber to innervate the muscle cells and to form functional in-vitro NMJs. Next we provide procedures to measure axon growth and to reliably quantify NMJ activity using imaging of both muscle contractions and fast intracellular calcium changes. This platform allows precise control, monitoring and manipulation of subcellular microenvironments. Specifically, it enables to distinguish local from retrograde signaling mechanisms and allows restricted experimental intervention in local compartments along the muscle-neuron route.
authors.affiliation: Department of Physiology and Pharmacology, Sackler Faculty of Medicine, and the Sagol School of Neuroscience, Tel Aviv University, Tel Aviv 69978, Israel.
authors.forename: Ariel
authors.initials: A
authors.lastname: Ionescu
authors.affiliation: Department of Physiology and Pharmacology, Sackler Faculty of Medicine, and the Sagol School of Neuroscience, Tel Aviv University, Tel Aviv 69978, Israel.
authors.forename: Eitan Erez
authors.initials: EE
authors.lastname: Zahavi
authors.affiliation: Department of Physiology and Pharmacology, Sackler Faculty of Medicine, and the Sagol School of Neuroscience, Tel Aviv University, Tel Aviv 69978, Israel.
authors.forename: Tal
authors.initials: T
authors.lastname: Gradus
authors.affiliation: Department of Physiology and Pharmacology, Sackler Faculty of Medicine, and the Sagol School of Neuroscience, Tel Aviv University, Tel Aviv 69978, Israel.
authors.forename: Keren
authors.initials: K
authors.lastname: Ben-Yaakov
authors.affiliation: Department of Physiology and Pharmacology, Sackler Faculty of Medicine, and the Sagol School of Neuroscience, Tel Aviv University, Tel Aviv 69978, Israel. Electronic address: eranpe@post.tau.ac.il.
authors.forename: Eran
authors.initials: E
authors.lastname: Perlson
mesh_terms: D000818:Animals; D020013:Calcium Signaling; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D051379:Mice; D044085:Microfluidics / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods; D009046:Motor Neurons / Q000166:cytology / Q000378:metabolism / Q000502:physiology; D009469:Neuromuscular Junction / Q000254:growth & development / Q000378:metabolism* / Q000502:physiology; D013116:Spinal Cord / Q000166:cytology
keywords: Axonal transport; Microfluidic-chamber; Motoneuron; Neuromuscular Junction; Neurotrophic factors