Elsevier

Neuroscience

Volume 110, Issue 3, 20 March 2002, Pages 555-567
Neuroscience

Neurturin enhances the survival of axotomized retinal ganglion cells in vivo: combined effects with glial cell line-derived neurotrophic factor and brain-derived neurotrophic factor

https://doi.org/10.1016/S0306-4522(01)00557-7Get rights and content

Abstract

In the present study we localized glial cell line-derived neurotrophic factor (GDNF), and the high affinity receptor for GDNF (GFRα-1) in the rat retina. We also examined the effects of neurturin on the survival of axotomized retinal ganglion cells (RGCs) and compared neurturin-mediated RGC rescue to GDNF and brain-derived neurotrophic factor (BDNF) neuroprotection. We administered combined injections of neurturin with BDNF or GDNF in order to determine if these factors rescue RGCs by different mechanisms. GDNF immunoreactivity was localized to RGCs, photoreceptors, and retinal pigment epithelial cells. GFRα-1 immunoreactivity was localized to RGCs, Müller cells, and photoreceptors. RGC densities in control retinas decreased from the original value of 2481±121 (RGCs/mm2±S.D.) to 347±100 at 14 days post-axotomy. Neurturin treatment significantly increased RGC survival after axotomy (745±94) similar to GDNF (868±110). BDNF treatment resulted in higher RGC survival (1109±156) than either neurturin or GDNF. Combined administration of neurturin with BDNF had additive effects on the survival of axotomized RGCs (1962±282), similar to combined administration of GDNF and BDNF (1825±269). Combined administration of neurturin and GDNF (1265±178) had an enhanced effect on RGC survival.

These results suggest that neurturin, GDNF, and BDNF act independently to rescue injured RGCs. Our results also suggest that RGCs and retinal Müller cells may be responsive to GDNF because they both express GFRα-1. The present findings have implications for the rescue of injured retinal ganglion cells, as well as other CNS neurons that are responsive to neurturin, GDNF, and BDNF, including midbrain dopaminergic neurons and motor neurons.

Section snippets

Intraorbital optic nerve transection and retrograde labeling

Adult, female Sprague–Dawley rats (225–250 g; Charles River), which were housed in a Level B pathogen-free environment, were used in all experiments. All efforts were made to minimize animal suffering and to use only the number of animals necessary to produce reliable scientific data. Experimental procedures were carried out according to the guidelines of the Canadian Council on Animal Care. Rats were anesthetized with i.p. injections of 7% chloral hydrate (0.42 mg/g of body weight) during

Localization of GDNF in the mammalian retina

GDNF mRNA has previously been localized in the retina (Nosrat et al., 1996). In order to determine which retinal cells express GDNF, GDNF immunohistochemistry was carried out on transverse retinal sections. GDNF was localized to cells in the ganglion cell layer, photoreceptor outer segments, and the retinal pigment epithelium (Fig. 1A). Intense GDNF immunoreactivity was also observed in RGC axon fascicles (Fig. 1B). In order to determine if the GDNF immunoreactive cells in the ganglion cell

Discussion

In the present study we localized GDNF and its high affinity receptor GFRα-1 in the mammalian retina. We also demonstrated that neurturin has neurotrophic effects on axotomized RGCs in vivo. Furthermore, combined administration of neurturin with GDNF produces enhanced RGC survival after axotomy. This enhanced effect was not as striking as the additive RGC survival observed when either neurturin or GDNF was combined with BDNF. These findings suggest that neurturin, GDNF, and BDNF may act through

Conclusion

The results of the present study suggest that RGCs and Müller cells express GFRα-1, the high affinity receptor for GDNF. RGCs may be directly responsive to GDNF, since previous findings suggest that RGCs express Ret. We have also shown for the first time that neurturin protects injured RGCs following optic nerve transection. Combined administration of neurturin with GDNF or BDNF is more effective in enhancing the survival of axotomized RGCs than intraocular administration of each factor

Acknowledgements

This work was supported by Natural Sciences and Engineering Research Council of Canada Operating Grant OGP0171190 to A.K.B. and an NSERC studentship to P.D.K. P.D.K. is supported by the Ontario Neurotrauma Foundation.

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