N1-Src kinase is required for primary neurogenesis in Xenopus tropicalis

Philip Alexander Lewis, Isobel Claire Bradley, Alastair Robert Pizzey, Harry Victor Isaacs, Gareth John Owen Evans

Research output: Contribution to journalArticlepeer-review

Abstract

The presence of the neuronal-specific N1-Src splice variant of the C-Src tyrosine kinase is conserved through vertebrate evolution, suggesting an important role in complex nervous systems. Alternative splicing involving a N1-Src specific microexon leads to a five or six amino acid insertion into the SH3 domain of Src. A prevailing model suggests that N1-Src regulates neuronal differentiation via cytoskeletal dynamics in the growth cone. Here we have investigated the role of n1-src in the early development of the amphibian Xenopus tropicalis, and find that n1-src expression is regulated in embryogenesis, with highest levels detected during the phases of primary and secondary neurogenesis. In situ hybridisation analysis, using locked nucleic acid (LNA) oligo probes complementary to the n1-src microexon indicate that n1-src expression is highly enriched in the open neural plate during neurula stages and in the neural tissue of adult frogs. Given the n1-src expression pattern, we investigated a possible role for n1-src in neurogenesis. Using splice site-specific antisense morpholino oligos, we are able to inhibit n1-src splicing, whilst preserving c-src expression. Differentiation of neurons in the primary nervous system is reduced in n1-src knockdown embryos, accompanied by a severely impaired touch response in later development. These data reveal an essential role for n1-src in amphibian neural development and suggest that alternative splicing of C-Src in the developing vertebrate nervous system evolved to regulate neurogenesis.
Original languageEnglish
Pages (from-to)8477-8584
Number of pages8
JournalJournal of neuroscience
Volume37
Issue number35
Early online date1 Aug 2017
DOIs
Publication statusPublished - 30 Aug 2017

Bibliographical note

© 2017 Lewis, Bradley et al.

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