Identification of Dp71 Isoforms Expressed in PC12 Cells: Subcellular Localization and Colocalization with β-Dystroglycan and α1-Syntrophin

J Mol Neurosci. 2016 Feb;58(2):201-9. doi: 10.1007/s12031-015-0657-8. Epub 2015 Sep 28.

Abstract

Several dystrophin Dp71 messenger RNA (mRNA) alternative splice variants have been described. According to the splicing of exon 78 or intron 77, Dp71 proteins are grouped as Dp71d, Dp71f, and Dp71e, and each group has a specific C-terminal end. In this study, we explored the expression of Dp71 isoforms at the complementary DNA (cDNA) level and the subcellular localization of recombinant Myc-Dp71 proteins in PC12 cells. We determined that PC12 cells express Dp71a, Dp71c, Dp71ab, Dp71e, and Dp71ec mRNA splice variants. In undifferentiated and nerve growth factor-differentiated PC12 Tet-ON cells, Dp71a, Dp71ab, and Dp71e were found to localize and colocalize with β-dystroglycan and α1-syntrophin in the periphery/cytoplasm, while Dp71c and Dp71ec were mainly localized in the cell periphery and showed less colocalization with β-dystroglycan and α1-syntrophin. The levels of Dp71a, Dp71e, and Dp71ec were increased in the nucleus of differentiated PC12 Tet-ON cells compared to undifferentiated cells. Dp71 isoforms were also localized in neurite extensions and growth cones.

Keywords: Dp71 isoforms; PC12 Cells; Subcellular localization; α1-Syntrophin; β-Dystroglycan.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium-Binding Proteins / genetics
  • Calcium-Binding Proteins / metabolism*
  • Cell Nucleus / metabolism
  • Cytoplasm / metabolism
  • Dystroglycans / genetics
  • Dystroglycans / metabolism*
  • Dystrophin / genetics
  • Dystrophin / metabolism
  • Growth Cones / metabolism
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Muscle Proteins / genetics
  • Muscle Proteins / metabolism*
  • PC12 Cells
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • Protein Transport
  • Rats

Substances

  • Calcium-Binding Proteins
  • Dystrophin
  • Membrane Proteins
  • Muscle Proteins
  • Protein Isoforms
  • apo-dystrophin 1
  • syntrophin alpha1
  • Dystroglycans