Disabled-1 alternative splicing in human fetal retina and neural tumors

PLoS One. 2011;6(12):e28579. doi: 10.1371/journal.pone.0028579. Epub 2011 Dec 6.

Abstract

Background: The Reelin-Dab1 signaling pathway plays a critical role in the positioning of migrating neurons, dendrite formation and lamination in the developing central nervous system. We have previously identified two alternatively spliced forms of Dab1 in the developing chick retina: an early form, Dab1-E, expressed in retinal progenitor cells, and a late form, Dab1 or Dab1-L, expressed in amacrine and ganglion cells. Compared to Dab1-L, Dab1-E lacks two exons that encode two Src family kinase (SFK) phosphorylation sites.

Principal findings: Both Dab1-L and Dab1-E-like transcripts were identified in human fetal retina. Expression of human Dab1-L in primary chick retinal cultures resulted in Reelin-mediated induction of SFK phosphorylation and formation of neurite-like processes. In contrast, human Dab1-E-expressing cells retained an undifferentiated morphology. The human Dab1 gene is located within a common fragile site, and it has been postulated that it may function as a tumor suppressor. Analysis of Dab1 splice forms in retinoblastoma and neuroblastoma tumor cells revealed relative enrichment of Dab1-L-like (includes exons 7 and 8) and Dab1-E-like (excludes exons 7 and 8) transcripts in retinoblastoma and neuroblastoma, respectively. Treatment of retinoblastoma cell line RB522A with Reelin resulted in increased tyrosine phosphorylation of Dab1. As Nova2 has previously been implicated in the exclusion of exons 9B and 9C in Dab1, we examined the expression of this splicing factor in neuroblastoma and retinoblastoma cell lines. Nova2 was only detected in neuroblastoma cells, suggesting a correlation between Nova2 expression and increased levels of Dab1-E-like splice forms in neuroblastoma.

Conclusions: These results indicate that alternative splicing of Dab1 is conserved in avian and mammalian species, with Dab1-L driving SFK phosphorylation in both species. Dab1-E- and Dab-L-like isoforms are also expressed in childhood neural tumors, with preferential enrichment of Dab1-L-like and Dab1-E-like isoforms in retinoblastoma and neuroblastoma, respectively.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / genetics*
  • Adaptor Proteins, Signal Transducing / metabolism*
  • Alternative Splicing*
  • Animals
  • Base Sequence
  • Central Nervous System / embryology
  • Chick Embryo
  • Exons
  • Humans
  • Microscopy, Fluorescence / methods
  • Molecular Sequence Data
  • Nerve Tissue Proteins / genetics*
  • Nerve Tissue Proteins / metabolism*
  • Nervous System Neoplasms / metabolism*
  • Neuroblastoma / metabolism
  • Phosphorylation
  • Polymerase Chain Reaction / methods
  • Protein Isoforms
  • Reelin Protein
  • Retina / embryology*
  • Retina / metabolism*
  • Retinoblastoma / metabolism
  • Sequence Homology, Nucleic Acid
  • src-Family Kinases / metabolism

Substances

  • Adaptor Proteins, Signal Transducing
  • DAB1 protein, human
  • Nerve Tissue Proteins
  • Protein Isoforms
  • Reelin Protein
  • src-Family Kinases
  • RELN protein, human