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Age-related changes underlie switch in netrin-1 responsiveness as growth cones advance along visual pathway

Abstract

Retinal axons are led out of the eye by netrin-1, an attractive guidance cue which is secreted at the optic nerve head. In the optic pathway, however, netrin-1 is expressed in areas that exclude retinal axon growth. This suggests that axons may change in their responsiveness to netrin-1 as they advance along the pathway. Indeed, in our 'whole-pathway' preparation in Xenopus, a gradual change from attraction to repulsion occurred as retinal axons emerged from progressively distal points along the pathway. We also found that axons that were aged in culture without pathway experience underwent a similar change, which correlated with a decline in cyclic AMP (cAMP) and netrin-1 receptor expression. Cyclic AMP elevators and adenosine A2b receptor agonists rejuvenated the behavior of old growth cones, causing them to regain attraction to netrin-1, whereas antagonists caused young growth cones to be repelled. These findings show that netrin-1 responsiveness is developmentally regulated and suggest that intrinsic changes that lower cAMP levels underlie this regulation.

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Figure 1: Netrin-1 expression in the developing optic pathway.
Figure 2: Preparation of whole pathway explants.
Figure 3: Effect of netrin-1 on growth cones at progressively advanced positions along the optic pathway.
Figure 4: Endogenous cAMP activity declines with age.
Figure 5: Developmental downregulation of DCC and A2b receptors in retinal growth cones.
Figure 6: Netrin-1 responsiveness changes after manipulation of netrin receptors.

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Acknowledgements

We thank A. Wiemelt for the gift of cAMP antibody, M. Tessier-Lavigne for providing netrin-1 and useful discussions, P. Mehlen for DCC protein and J. Skepper for confocal imaging. We also thank S. Hopper for technical assistance and W. Harris for many helpful discussions. This work was funded by a Medical Research Council (UK) Programme Grant (to C.E.H.).

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Correspondence to C.E. Holt.

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Shewan, D., Dwivedy, A., Anderson, R. et al. Age-related changes underlie switch in netrin-1 responsiveness as growth cones advance along visual pathway. Nat Neurosci 5, 955–962 (2002). https://doi.org/10.1038/nn919

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