Characterization of Dp71Δ_78‐79, a novel dystrophin mutant that stimulates PC12 cell differentiation

Abstract

J. Neurochem. (2011) 119, 697–707. Abstract Dp71 has an important role in the central nervous system. To better understand the function of Dp71 domains in neuronal differentiation, PC12 cells were stably transfected with a dystrophin mutant, Dp71Δ78‐79, which lacks exons 78 and 79. Based on the percentage of cells bearing neurites and neurite length analyses, we found that cells stably expressing Dp71Δ78‐79 (PC12‐C11) differentiate more efficiently than non‐transfected cells. While wild‐type cells reach their maximum differentiation 9–12 days after initiating the differentiation process, the PC12‐C11 cells reach differentiation in 4–6 days. Protein expression analysis showed a down‐regulation of Dp71a and an up‐regulation of Dp71ab and/or Up71, β‐dystroglycan and neuron‐specific enolase in undifferentiated and in neural growth factor differentiated PC12‐C11 cells. No change was observed in the expression of Grb2 and Up400. The subcellular localization of Dp71Δ78‐79 was in the cell periphery, and there was no change in localization during the differentiation process, which was also observed throughout the neurite extensions.

Altered Biological Properties in Dp71 Over-Expressing HBE Cells

“…Via associating with dystrophin associated protein complex, dystrophin was proved to play important roles in various organs [16, 17]. Via up-regulation of FAK in the cytoplasmic DAPCs and lamin B1 in the nuclues from the transcription level, the HBE-Dp71d and HBE-Dp71f cells altered their biological traits in several different aspects.…”

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

“…All medium supplements were obtained from Gibco (Gibco, Rockville, MD, USA). PC12 and HeLa cells were cultured as previously described (Aragon et al 2011).…”

“…The expression of Myc-Dp71 proteins in PC12 Tet-ON cells was induced with 1 μg/ml doxycycline for 24 h and throughout the differentiation process. PC12 Tet-ON cells were differentiated with 50 ng/ml of NGF (Gibco) in growth medium, and PC12 cells were differentiated as previously described (Aragon et al 2011). The differentiation medium was replaced every 3 days.…”

“…The three Dp71 isoform groups have unique C-terminal ends (Fig. 1), and to date, reported data have suggested that the C-terminus of Dp71 is involved in its specific functions (Aleman et al 2001;Szabó et al 2004;Aragon et al 2011). Dp71 has been reported to have roles in cell division and membrane stabilization.…”

“…Furthermore, nuclear dystrophin-associated proteins (DAPs) and nuclear matrix components have been reported to have roles in cell membrane morphology maintenance, the adipogenesis process, and the homeostasis and clustering of potassium and water channels (Tadayoni et al 2012). Dp71 has also been reported to have a role in the neuronal differentiation of PC12 (Aragon et al 2011) andN1E-115 cells (Morales-Lázaro et al 2010).…”