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  • Original Paper
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Strict regulation of c-Raf kinase levels is required for early organogenesis of the vertebrate inner ear

Oncogene volume 18, pages 429–437 (1999)Cite this article

Abstract

Regulation of organogenesis involves a dynamic balance of the mechanisms regulating cell division, differentiation and death. Here we have investigated the pattern of expression of c-Raf kinase in the inner ear during early developmental stages and the consequences of manipulating c-Raf levels by misexpression of c-raf viral vectors in organotypic cultures of otic vesicle explants. We found that otic vesicles expressed c-Raf and its level remained constant during embryonic days 2 and 3 (E2-E3). c-Raf activity was increased in response to insulin like growth factor-I (IGF-I) and the activation by IGF-I of the c-Raf kinase pathway was a requirement to turn on cell proliferation in the otic vesicle. Overexpression of c-raf in E2.5 explants increased the proliferative response to low serum and IGF-I and blocked differentiation induced by retinoic acid. The increase in c-Raf levels also prevented nerve growth factor (NGF)-dependent induction of programmed cell death. Consistent with these results, the expression of a dominant negative c-Raf mutant potentiated retinoic acid action and decreased the rate of cell proliferation. We conclude that a strict control of c-Raf levels is essential for the co-ordination of the biological processes that operate simultaneously during early inner ear development.

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Abbreviations

(CEF):

Chicken embrionary fibroblasts

(CVG):

cochleovestibular ganglion

(E):

embryonic day

(IGF-I):

insulin-like growth factor-I

(MAP kinase):

mitogen activated protein kinase

(NGF):

nerve growth factor

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Acknowledgements

YL held a CSIC research contract and CS was supported by Ministerio de Education y Cultura. We wish to thank Drs Stephen Hughes, Volker M Vogt and Luis Alvarez for their generous help and the continuos technical advice provided along this work, and to Drs Luis Alvarez, Fernando Giraldez, Flora de Pablo, Jaime Renart, David Jones and Carmen G Vallejo, for critical reading of the manuscript. We also thank Antonio Fernández and Javier Pérez for photographic work and illustrations respectively. The research was supported by grants from Dirección General Investigación Ciencia y Tecnología (PM96-0075) and Europharma (Boehringer Ingelheim Inc.) to I V-N and Deutsche Forschungsgemeinschaft (DFG) Sonderforschungsbereiche (SFB) 465 and 172 to URR.

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  1. Instituto de Investigaciones Biomédicas, Consejo Superior de Investigaciones Científicas (CSIC), Arturo Duperier 4, 28029, Madrid, Spain

    Carmen Sanz, Yolanda León & Isabel Varela-Nieto

  2. Institut für Medizinische Strahlenkunde und Zellforschung (MSZ), Bayerische Julius-Maximillians-Universität, Versbacher Str. 5, 97078, Würzburg, Germany

    Jakob Troppmair & Ulf R Rapp

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  1. Carmen Sanz
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Sanz, C., León, Y., Troppmair, J. et al. Strict regulation of c-Raf kinase levels is required for early organogenesis of the vertebrate inner ear. Oncogene 18, 429–437 (1999). https://doi.org/10.1038/sj.onc.1202312

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