Characterization of Netrin-1, Neogenin and cUNC-5H3 expression during chick dorsal root ganglia development

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Abstract

The molecular mechanisms that control the axon pathfinding of different subtypes of dorsal root ganglia (DRG) neurons are poorly understood. To address whether Netrin-1-Neogenin/UNC-5 signaling contributes to sensory axon pathfinding, we cloned chick UNC5 homolog 3 (cUNC-5H3) and characterized the spatial, cellular and temporal expression patterns of Netrin-1, Neogenin and cUNC-5H3 in cutaneous and proprioceptive DRG neurons. We have found that Netrin-1 is only expressed by late-arising cutaneous neurons. In contrast, cUNC-5H3 expression is restricted to proprioceptive neurons. Neogenin is expressed in all DRG neurons. Our results indicate that the pathfinding molecules, Netrin-1 and cUNC-5H3 are differentially expressed by early cutaneous and proprioceptive neurons.

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Results and discussion

Here we examined components of the Netrin-1 signaling pathway during chick DRG development. Netrin-1 is a bifunctional guidance cue and the colorectal cancer (DCC)/Neogenin (Vielmetter et al., 1994; Keino-Masu et al., 1996; Kolodziej et al., 1996) and UNC-5 family of receptors (Hedgecock et al., 1990; Leonardo et al., 1997; Colavita and Culotti, 1998) mediate attractive and repulsive responses to Netrin-1, respectively.

Experimental procedures

cUNC-5H3 was cloned using RACE (Rapid amplification of cDNA ends, Frohman, 1990), RT–PCR and degenerate primers: (sense) 5′-AARTAYCARGARATHCCNTTYTA-3′, (anti-sense) 5′-GCYTCCCANARRTCNARDATNAC-3′. Phylogenetic analysis was done using CLUSTAL W (1.81) software (http://www.ebi.ac.uk/clustalw/). Chick DRGs at indicated stages (Hamburger and Hamilton, 1951) were dissected and cultured on non-adherent dishes, either in the absence (Fig. 5) or presence (Fig. 4) of neurotrophins (Chemicon). mRNA was

Acknowledgements

This work was supported by NIH grant R01 NS38138 to M.L.C.

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