Elsevier

Neuroscience

Volume 109, Issue 4, 22 February 2002, Pages 643-656
Neuroscience

Expression of netrin-1 and netrin-1 receptor, DCC, in the rat olfactory nerve pathway during development and axonal regeneration

https://doi.org/10.1016/S0306-4522(01)00535-8Get rights and content

Abstract

Netrin-1 is a bifunctional secreted protein that directs axon extension in various groups of developing axonal tracts. The transmembrane DCC (deleted in colorectal cancer) receptor is described as netrin-1 receptor and is involved in the attractive effects of netrin-1. In this study, we examined the spatio-temporal expression patterns of both netrin-1 and DCC in the rat olfactory system at different stages of development and during axonal regeneration following unilateral bulbectomy. High DCC expression was detected on the pioneer olfactory axons as they are extending toward the telencephalon. This expression was transient since from embryonic day 16 onwards, DCC was no longer detected along the olfactory nerve path. From embryonic day 14 until birth, DCC was also expressed within the mesenchyme surrounding the olfactory epithelium. During the same period, netrin-1 protein was detected along the trajectory of olfactory axons up to the olfactory bulb and its expression pattern in the nasal mesenchyme largely overlapped that of DCC. Moreover, netrin-1 continued to be present during the two first post-natal weeks, and a weak protein expression still persisted in the dorso-medial region of the olfactory epithelium in adult rats. While unilateral bulbectomy induced a transient up-regulation of netrin-1 in the lamina propria, particularly in the dorso-medial region of the neuroepithelium, no DCC expression was detected on the regenerating olfactory axons. In the developing olfactory bulb, the extension of mitral cell axons was associated with DCC presence while netrin-1 was absent along this axonal path. DCC was also highly expressed in the newly formed glomeruli after birth, and a weak DCC expression was still detected in the glomerular layer in adult rats.

Taken together, these data support the notion that netrin-1, via DCC expressed on axons, may play a role in promoting outgrowth and/or guidance of pioneering olfactory axons toward the olfactory bulb primordium. Moreover, association of netrin-1 with mesenchymal DCC may provide a permissive environment to the growth of both pioneer and later-growing axons. The maintenance of netrin-1 expression in the nasal mesenchyme of adult rats as well as its regional up-regulation following unilateral bulbectomy infer that netrin-1, even in the absence of DCC, may be involved in the process of axonal growth of newly differentiated olfactory receptor neurons probably through the use of other receptors.

Section snippets

Animals

Wistar SPF (IFFA-Credo, France) rat embryos from the 13th to the 19th day of gestation (day of conception=E1) and rats aged 1, 6, 15, 30 and 50 post-natal days (day of birth=P1) were used in this study. Animals were housed in polypropylene cages, kept in a 12–12-h light–dark cycle and provided with food and water ad libitum.

Surgical procedures

Thirty-day-old male rats were anesthetized with Equithesin (3 mg/kg, i.p.). Animals were placed in a stereotaxic apparatus and the frontal bones over the olfactory bulbs

Developmental regulation of DCC expression

The two monoclonal antisera used to determine the expression of DCC in the developing olfactory system gave very similar patterns of immunoreactivity. Since a weaker labelling intensity was generally observed with AF5 antibody generated against the extracellular domain of the protein, we therefore reported results obtained with the antibody directed against the intracellular domain of DCC.

Netrin-1 and DCC expressions in the developing olfactory pathway

One of the netrin-1 receptors, DCC, is known to be expressed on various projecting populations of axons during early stages of the neural development (Keino-Masu et al., 1996, Deiner et al., 1997, De la Torre et al., 1997, Shu et al., 2000, Anderson et al., 2000). In agreement with these studies, our results demonstrate that DCC protein is dynamically regulated on the axon populations emerging from the olfactory epithelium and the vomeronasal epithelium during embryogenesis and early post-natal

Conclusion

The present study provides a detailed analysis of the spatio-temporal expression patterns of both netrin-1 and DCC proteins in the developing rat olfactory system. It has been shown that the association of netrin-1 expression along DCC-expressing olfactory axons is restricted to the initial period of olfactory nerve pathfinding, which suggests that netrin-1 may play a role in the directed outgrowth of the nascent olfactory axons toward the telencephalon. The idea that this function should be

Acknowledgements

This study was supported by the Centre National de la Recherche Scientifique (UMR 5020, UMR 5534), the Ligue contre le cancer (P.M.), the ARC (5931, P.M.) and the University of Lyon (BQR).

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