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Distribution of GAP-43, β-III tubulin and F-actin in developing and regenerating axons and their growth cones in vitro, following neurotrophin treatment

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Journal of Neurocytology

Abstract

Brain derived neurotrophic factor (BDNF) when added to explant cultures of both embryonic and adult retinal ganglion cell (RGC) axons exerted a marked effect on their growth cone size and complexity and also on the intensity of GAP-43, ß-III tubulin and F-actin immunoreaction product in their axons. GAP-43 was distributed in axons, lamellipodia, and filopodia whereas ß-III tubulin was distributed along the length of developing and adult regenerating axons and also in the C-domain of their growth cones. BDNF-treated developing RGC growth cones were larger and displayed increased numbers of GAP-43 and microtubule-containing branches. Although filopodia and lamellipodia were lost from both developing and adult RGC growth cones following trkB-IgG treatment, the intensity of the immunoreaction product of all these molecules was reduced and trkB-IgGs had no effect on the axonal distribution of ß-III tubulin and GAP-43. BDNF-treated growth cones also displayed increased numbers of F-actin containing filopodia and axonal protrusions. This study demonstrates, for the first time, that trkB-IgG treatment causes the loss of F-actin in the P-domain of growth cone tips in developing and regenerating RGC axons. Although microtubules and F-actin domains normally remained distinct in cultured growth cones, ß-III tubulin and F-actin overlapped within the growth cone C-domain, and within axonal protrusions of adult RGC axons, under higher concentrations of BDNF. The collapse of RGC growth cones appeared to correlate with the loss of F-actin. In vitro, trkB signalling may therefore be involved in the maintenance and stabilisation of RGC axons, by influencing F-actin polymerisation, stabilisation and distribution.

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Authors and Affiliations

  1. Department of Anatomy and Developmental Biology, University College London, Gower Street, WC1E 6BT, London

    Ovokeloye Avwenagha & Gregor Campbell

  2. Education Directorate, Institute of Pathology, Barts and the London, Whitechapel, E1 1BB, London

    Margaret M. Bird

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  1. Ovokeloye Avwenagha
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  2. Gregor Campbell
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Correspondence to Margaret M. Bird.

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Avwenagha, O., Campbell, G. & Bird, M.M. Distribution of GAP-43, β-III tubulin and F-actin in developing and regenerating axons and their growth cones in vitro, following neurotrophin treatment. J Neurocytol 32, 1077–1089 (2003). https://doi.org/10.1023/B:NEUR.0000021903.24849.6c

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