title: Can hippocampal neurites and growth cones climb over obstacles?
abstract: Guidance molecules, such as Sema3A or Netrin-1, can induce growth cone (GC) repulsion or attraction in the presence of a flat surface, but very little is known of the action of guidance molecules in the presence of obstacles. Therefore we combined chemical and mechanical cues by applying a steady Netrin-1 stream to the GCs of dissociated hippocampal neurons plated on polydimethylsiloxane (PDMS) surfaces patterned with lines 2 µm wide, with 4 µm period and with a height varying from 100 to 600 nm. GC turning experiments performed 24 hours after plating showed that filopodia crawl over these lines within minutes. These filopodia do not show staining for the adhesion marker Paxillin. GCs and neurites crawl over lines 100 nm high, but less frequently and on a longer time scale over lines higher than 300 nm; neurites never crawl over lines 600 nm high. When neurons are grown for 3 days over patterned surfaces, also neurites can cross lines 300 nm and 600 nm high, grow parallel to and on top of these lines and express Paxillin. Axons - selectively stained with SMI 312 - do not differ from dendrites in their ability to cross these lines. Our results show that highly motile structures such as filopodia climb over high obstacle in response to chemical cues, but larger neuronal structures are less prompt and require hours or days to climb similar obstacles.
authors.affiliation: Neurobiology Sector, International School for Advanced Studies (SISSA), Trieste, Italy.
authors.forename: Thuy Linh
authors.initials: TL
authors.lastname: Lien
authors.forename: Jelena
authors.initials: J
authors.lastname: Ban
authors.forename: Massimo
authors.initials: M
authors.lastname: Tormen
authors.forename: Elisa
authors.initials: E
authors.lastname: Migliorini
authors.forename: Gianluca
authors.initials: G
authors.lastname: Grenci
authors.forename: Alessandro
authors.initials: A
authors.lastname: Pozzato
authors.forename: Vincent
authors.initials: V
authors.lastname: Torre
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D018929:Cell Culture Techniques; D020439:Growth Cones / Q000502:physiology*; D006624:Hippocampus / Q000502:physiology*; D009414:Nerve Growth Factors / Q000378:metabolism; D000075388:Netrin-1; D016501:Neurites / Q000502:physiology*; D009474:Neurons / Q000502:physiology; D051419:Paxillin / Q000378:metabolism; D051381:Rats; D014404:Tubulin / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000378:metabolism
chemical_list: D009414:Nerve Growth Factors; C000617354:Ntn1 protein, rat; D051419:Paxillin; D014404:Tubulin; D025521:Tumor Suppressor Proteins; D000075388:Netrin-1
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