RRC ID 61352
Author Lin L, Isacson O.
Title Axonal growth regulation of fetal and embryonic stem cell-derived dopaminergic neurons by Netrin-1 and Slits.
Journal Stem Cells
Abstract The physical restoration of dopamine circuits damaged or lost in Parkinson disease by implanting embryonic stem (ES)-derived cells may become a treatment. It is critical to understand responses of ES-derived dopamine (DA) neurons to guidance signals that determine axonal path and targeting. Using a collagen gel culture system, we examined effects of secreted molecules Netrin-1 and Slits on neurite outgrowth of fetal DA neurons and murine ES-differentiated DA neurons. We have previously shown that fetal DA neurons express DCC and Robo1/2 receptors and that Netrin-1 and Slit2 function as an attractant and a repellent for DA neurite outgrowth. In the present study, we observe that both Slit1 and Slit3 repel and inhibit neurite growth of fetal DA neurons. Here, we also demonstrate that ES-differentiated neurons including DA neurons express the Netrin receptor DCC and Slit receptor Robo proteins. In the gel culture system of ES cells, Netrin-1 promoted neurite outgrowth mediated by DCC receptor, and Slit1 and Slit3 were inhibitory for neurite outgrowth through Robo receptors. Slit2 appeared to exert inhibitory as well as repulsive effects in the coculture assay. However, unlike fetal DA neurites, no directed neurite outgrowth was observed in the cocultures of ES-derived DA neurons with Netrin-1-, Slit1-, and Slit3-producing cells. The findings suggest that ES-derived DA neurons generated by current protocols can respond to guidance cues in vitro in a similar manner to fetal cells but also exhibit distinct responses. This may result from developmental differences generated by present in vitro methods of cell patterning or conditioning during ES cell differentiation.
Volume 24(11)
Pages 2504-13
Published 2006-11-1
DOI 10.1634/stemcells.2006-0119
PII 2006-0119
PMID 16840550
PMC PMC2613222
MeSH Animals Axons / metabolism* Cell Line Coculture Techniques DCC Receptor Dopamine / metabolism* Embryonic Stem Cells / metabolism* Fetal Stem Cells / metabolism* Fluorescent Antibody Technique Humans Intercellular Signaling Peptides and Proteins / genetics Intercellular Signaling Peptides and Proteins / metabolism* Membrane Proteins / genetics Membrane Proteins / metabolism* Mice Nerve Growth Factors / genetics Nerve Growth Factors / metabolism* Nerve Tissue Proteins / genetics Nerve Tissue Proteins / metabolism* Netrin-1 Neurites / metabolism Rats Rats, Sprague-Dawley Receptors, Cell Surface / metabolism Receptors, Immunologic / metabolism Transfection Tumor Suppressor Proteins / genetics Tumor Suppressor Proteins / metabolism*
IF 6.022
Human and Animal Cells MC3T3-G2/PA6(RCB1127)