title: Coculture system of keratinocytes and dorsal-root-ganglion-derived cells for screening neurotrophic factors involved in guidance of neuronal axon growth in the skin.
abstract: The density of peripheral nerve fibres is increased in atopic dermatitis. Moreover, reduction in the fibres in a mouse model of atopic dermatitis reduces scratching behaviour. Thus, regulation of nerve fibre extension could be an effective strategy to reduce itching in pruritus dermatosis. In this study, we established a new coculture system of keratinocytes and dorsal-root-ganglion-derived cells using an apparatus, AXIS(™) , which consists of two different channels connected via a set of microgrooves, through which signalling molecules and axons, but not living cells, can pass. When we seeded keratinocytes in one chamber, extension of nerve fibres was observed from dorsal root ganglion cells seeded in the other chamber. Addition of anti-BDNF antibody in the keratinocyte-seeded chamber significantly reduced the extension. Application of Semaphorin 3A also reduced the extension by approximately 50%. We suggest that this coculture system may be useful for screening of anti-itching drugs.
authors.affiliation: CREST, Japan Science and Technology Agency, Tokyo, Japan.
authors.forename: Jun-Ichi
authors.initials: J
authors.lastname: Kumamoto
authors.forename: Masashi
authors.initials: M
authors.lastname: Nakatani
authors.forename: Moe
authors.initials: M
authors.lastname: Tsutsumi
authors.forename: Makiko
authors.initials: M
authors.lastname: Goto
authors.forename: Sumiko
authors.initials: S
authors.lastname: Denda
authors.forename: Kentaro
authors.initials: K
authors.lastname: Takei
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Denda
mesh_terms: D000818:Animals; D000982:Antipruritics / Q000494:pharmacology; D001369:Axons / Q000187:drug effects / Q000648:ultrastructure; D019208:Brain-Derived Neurotrophic Factor / Q000037:antagonists & inhibitors; D018920:Coculture Techniques / Q000379:methods; D003876:Dermatitis, Atopic / Q000188:drug therapy; D004353:Drug Evaluation, Preclinical; D005727:Ganglia, Spinal / Q000166:cytology* / Q000187:drug effects* / Q000254:growth & development; D006801:Humans; D015603:Keratinocytes / Q000166:cytology* / Q000187:drug effects*; D051379:Mice; D009412:Nerve Fibers / Q000187:drug effects / Q000648:ultrastructure; D009414:Nerve Growth Factors / Q000494:pharmacology*; D010525:Peripheral Nerves / Q000187:drug effects / Q000254:growth & development; D040121:Semaphorin-3A / Q000494:pharmacology; D012867:Skin / Q000187:drug effects / Q000293:injuries
chemical_list: D000982:Antipruritics; D019208:Brain-Derived Neurotrophic Factor; D009414:Nerve Growth Factors; D040121:Semaphorin-3A
keywords: DRG derived neuron; axon growth; coculture; keratinocyte; neurotrophin