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The Journal of Neuroscience, September 22, 2004, 24(38):8346-8353; doi:10.1523/JNEUROSCI.1809-04.2004

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Development/Plasticity/Repair
Role of Purpurin as a Retinol-Binding Protein in Goldfish Retina during the Early Stage of Optic Nerve Regeneration: Its Priming Action on Neurite Outgrowth

Toru Matsukawa,¹ Kayo Sugitani,^1,2 Kazuhiro Mawatari,² Yoshiki Koriyama,¹ Zhongwu Liu,¹ Masayuki Tanaka,¹ and Satoru Kato¹

¹Department of Molecular Neurobiology, Graduate School of Medicine, and ²School of Health Sciences, Faculty of Medicine, Kanazawa University, Kanazawa, Ishikawa 920-8640, Japan

Unlike mammals, the fish optic nerve can regenerate after injury. So far, many growth or trophic factors have been shown as an axon-regenerating molecule. However, it is totally unknown what substance regulates or triggers the activity of these factors on axonal elongation. Therefore, we constructed a goldfish retina cDNA library prepared from the retina treated with optic nerve transection 5 d previously, when it was just before regrowing optic axons after injury. A cDNA clone for goldfish purpurin for which expression was upregulated during the early stage of optic nerve regeneration was isolated from the retina cDNA library. Purpurin was discovered as a secretory retinol-binding protein in developing chicken retinas. Levels of purpurin mRNA and protein transiently increased and rapidly decreased 2–5 d and 10 d after axotomy, respectively. Purpurin mRNA was localized to the photoreceptor cells, whereas the protein was diffusely found in all of the retinal layers. A recombinant purpurin alone did not affect any change of neurite outgrowth in explant culture of the control retina, whereas a concomitant addition of the recombinant purpurin and retinol first induced a drastic enhancement of neurite outgrowth. Furthermore, the action of retinol-bound purpurin was effective only in the control (untreated) retinas but not in those primed (treated) with a previous optic nerve transection. Thus, purpurin with retinol is the first candidate molecule of priming neurite outgrowth in the early stage of optic nerve regeneration in fish.

Key words: goldfish; optic nerve regeneration; purpurin; retinol-binding protein; neurite outgrowth; retina

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Received May 11, 2004; revised July 29, 2004; revised August 5, 2004;

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