title: Tracking Quantum-Dot labeled neurotropic factors transport along primary neuronal axons in compartmental microfluidic chambers.
abstract: Neurons are highly polarized cells, with very long axons. Neurotrophic factors like the neuronal growth factor (NGF) are secreted from neuronal targets to promote neuron survival and proper function. These neurotrophic factors must undergo retrograde axonal transport towards the cell body, wherein they initiate signaling pathways important for neurons' various functions and overall health. This process of long-distance axonal signaling is conducted by the dynein motor protein, which transmits signaling endosomes of ligand-receptor complexes retrogradely along microtubule tracks. Here we describe step by step the use of polydimethylsiloxane (PDMS) compartmentalized microfluidic chambers for tracking axonal transport of trophic factors, with a focus on labeled NGF. We describe in detail how to fabricate the molds, assemble the PDMS platform, plate neurons and image, as well as analyze NGF transport along the axon. This method is useful for studying molecular communication mechanisms within the neuron's different compartments as well as between the neuron and its diverse microenvironments, both in health and under pathological conditions.
authors.affiliation: Department of Physiology and Pharmacology, Sackler Faculty of Medicine, and the Sagol School of Neuroscience, Tel Aviv University, Tel Aviv, Israel.
authors.forename: Shani
authors.initials: S
authors.lastname: Gluska
authors.affiliation: Department of Physiology and Pharmacology, Sackler Faculty of Medicine, and the Sagol School of Neuroscience, Tel Aviv University, Tel Aviv, Israel.
authors.forename: Michael
authors.initials: M
authors.lastname: Chein
authors.affiliation: Department of Physiology and Pharmacology, Sackler Faculty of Medicine, and the Sagol School of Neuroscience, Tel Aviv University, Tel Aviv, Israel.
authors.forename: Nimrod
authors.initials: N
authors.lastname: Rotem
authors.affiliation: Department of Physiology and Pharmacology, Sackler Faculty of Medicine, and the Sagol School of Neuroscience, Tel Aviv University, Tel Aviv, 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, Israel.
authors.forename: Eran
authors.initials: E
authors.lastname: Perlson
mesh_terms: D000818:Animals; D001370:Axonal Transport / Q000502:physiology*; D001369:Axons / Q000378:metabolism*; D002478:Cells, Cultured; D056445:Cytoplasmic Dyneins / Q000378:metabolism; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D004622:Embryo, Mammalian / Q000294:innervation; D005260:Female; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D044085:Microfluidics / Q000379:methods*; D020932:Nerve Growth Factor / Q000378:metabolism*; D045663:Quantum Dots / Q000378:metabolism*
chemical_list: D004129:Dimethylpolysiloxanes; C013830:baysilon; D020932:Nerve Growth Factor; D056445:Cytoplasmic Dyneins
keywords: Axonal transport; Microfluidic chambers; Neurotrophic factors; Retrograde transport; Signaling endosomes