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

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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.

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Acknowledgments

We thank Dr. Dominique Mornet for providing the antibodies JAF and C4, Dr. Manuel Hernandez for providing the antibody anti-β-actin, Ivan Galván for his assistance with confocal microscopy, Guadalupe Aguilar for performing DNA sequencing, Verónica Ramírez for her clerical support, and Adalberto Herrera and Clemencia Salas for their technical assistance.

This work was supported by CONACyT grants: CB-2009-127600 and SEP-CONACyT-ECOS-ANUIES M11-S02 B000/064/12 to C. Montañez; CONACyT doctoral scholarships: 173195 to A. Martínez-Herrera, 263361 to C. Azotla-Vilchis, 263733 to L. Siqueiros-Márquez, and 199918 to A. Herrera-Salazar; and a post-doctoral grant: 76927 to G. Soid-Raggi.

Conflict of Interest

The authors declare that they have no conflict of interest.

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Author notes

  1. José Romo-Yáñez

    Present address: Departamento de Genómica Humana, Instituto Nacional de Perinatología, C.P. 11000, México, D.F., Mexico

  2. Alma Herrera-Salazar

    Present address: Departamento de Infectología e Inmunología, Instituto Nacional de Perinatología, C.P. 11000, México, D.F., Mexico

Authors and Affiliations

  1. Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del IPN, Av. IPN No. 2508, San Pedro Zacatenco, C.P. 07360, México, D. F., Mexico

    Jorge Aragón, Alejandro Martínez-Herrera, José Romo-Yáñez, Víctor Ceja, Coztli Azotla-Vilchis, Lourdes Siqueiros-Márquez, Gabriela Soid-Raggi, Alma Herrera-Salazar & Cecilia Montañez

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  1. Jorge Aragón
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Correspondence to Cecilia Montañez.

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Aragón, J., Martínez-Herrera, A., Romo-Yáñez, J. et al. Identification of Dp71 Isoforms Expressed in PC12 Cells: Subcellular Localization and Colocalization with β-Dystroglycan and α1-Syntrophin. J Mol Neurosci 58, 201–209 (2016). https://doi.org/10.1007/s12031-015-0657-8

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