Biochemical and histochemical analysis of 71 kDa dystrophin isoform (Dp71f) in rat brain

García-Tovar, Carlos; Pérez, A; Luna, J.; Mena, Raúl; Osorio, Beatriz; Aleman, Victor; Mondragón, Ricardo; Mornet, Dominique; Rendón, Álvaro; Hernández, José Manuel

doi:10.1078/0065-1281-00591

Cited by 51 publications

(28 citation statements)

References 64 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Schofield et al (1994), on the other hand, already suggested that apodystrophin-1 (i.e., Dp71, the major cerebral dystrophin) mRNA could be associated to glial cells. Further investigations in cell cultures (Alemann et al 2001;Imamura and Ozawa 1998), detected the presence of Dp71 in astrocytes, and Garcia-Tovar et al (2001) supported this localization by the immunohistochemical detection of glial fibrillary acidic protein (GFAP).…”

Section: Introductionmentioning

confidence: 84%

“…Bars: 60 mm, 50 mm, 50 mm, and 12 mm, respectively (Fabbrizio et al 1994). In recent studies in cell cultures, the 5F3 antibody has labeled (1) GFAP-immunopositive astrocytes (Garcia-Tovar et al 2001), and (2) cells producing the mRNA splice variant responsible for the production of Dp71f (Marquez et al 2003, PC12 cells). Our double-labeling results with GFAP support the occurrence in astrocytes of Dp71f, but not Dp71d.…”

Section: Discussionmentioning

confidence: 99%

“…Western blot analysis of protein extracts (Blake et al 1999, Garcia-Tovar et al 2001Huard et al 1992;Jung et al 1991) revealed the presence of dystrophin-protein family members in almost all brain regions. Uchino et al (1994) demonstrated lack of dystrophins in several brain areas of Duchenne dystrophy patients in contrast to control persons.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Immunofluorescence mapping of dystrophin in the rat brain: astrocytes contain the splice variant Dp71f, but this is confined to subpopulations

et al. 2004

Self Cite

View full text Add to dashboard Cite

Dystrophins are membrane-associated actin-binding proteins, recognized at first in muscular dystrophies. In the brain the full-length Dp427 has been detected, as well as Dp140 and Dp71 of the shorter variants. Dp71 seems to be their major representative in the brain, and it occurs as splice variants, Dp71f and Dp71d. Dystrophins have been demonstrated mainly in neurons. In tissue cultures, the glial data, mainly in situ, are still insufficient. The present mapping study reveals the astroglial localization of the splice variant Dp71f, using a monoclonal antibody (5F3, developed by D. Mornet) specific for its additional 31 last amino acids. In parallel, another monoclonal antibody was used (Dys2, Novocastra) that detects the Dp71d, Dp427, as well as Dp140 and other short variants. Rats were overdosed with ether and perfused transcardially with 4% phosphate-buffered paraformaldehyde solution. Floating Vibratome sections were processed for immunohistochemical labeling with fluorescent secondary antibodies. In some animals the reactive glia were investigated following stab wounds in ketamine-xylazine anesthesia. Only the 5F3 antibody labeled astrocytes, however, not in general but in special localizations, mainly along the glia limitans of the pial surface, below the ependyma and in the reactive glia. Perivascular astrocytes were consistently labeled only where the vessels entered the brain, and in some circumventricular organs. The 5F3 antibody also labeled the ependyma and the residual subventricular zone. In contrast to the astrocytes, neurons were labeled throughout the brain. Dys2 antibody (to Dp71d and longer isoforms) labeled neurons in a distribution similar to that of 5F3, but rarely labeled astroglia and only in perivascular rings. Dp71f positivity seems to occur in those astrocyte populations that proved to be immunopositive to glial fibrillary acidic protein (GFAP) and produced laminin in former studies.

show abstract

Section: Introductionmentioning

confidence: 84%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Immunofluorescence mapping of dystrophin in the rat brain: astrocytes contain the splice variant Dp71f, but this is confined to subpopulations

et al. 2004

Self Cite

View full text Add to dashboard Cite

show abstract

“…The primary antibodies were anti-human p53 1:500 (Santa Cruz Biotechnology, Santa Cruz, CA), which recognizes both mutant and wild-type forms, anti-b galactosidase 1:500 (Promega) and anti-b actin 1:600. 28 …”

Section: Western Blot Analysismentioning

confidence: 99%

Targeted-simultaneous expression of Gas1 and p53 using a bicistronic adenoviral vector in gliomas

et al. 2007

View full text Add to dashboard Cite

The targeted expression of transgenes is one of the principal goals of gene therapy, and it is particularly relevant for the treatment of brain tumors. In this study, we examined the effect of the overexpression of human gas1 (growth arrest specific 1) and human p53 cDNAs, both under the transcriptional control of a promoter of the human glial fibrillary acidic protein (gfa2), employing adenoviral expression vectors, in glioma cells. We showed that the targeted overexpression of gas1 and p53 (AdSGas1 and AdSp53, respectively) in rat glioma cells (C6) reduced the number of viable cells and induced apoptosis. Moreover, the adenovirally targeted expression of these genes also reduced tumor growth in vivo. Unexpectedly, there was no additive effect when both gas1 and p53 were simultaneously expressed in the same cells using a bicistronic adenoviral vector. We suggest that Gas1 does not act in combination with p53 in the C6 and U373 glioma cell lines, inducing apoptosis and cell cycle arrest. Our results indicate that the targeted expression of tumor suppressor genes (gas1 and p53) regulated by the gfa2 promoter, together with adenoviral vectors may provide an interesting approach for adjuvant selective glioma gene therapy.

show abstract

“…10,12 The secondary antibody (Jackson ImmunoResearch, West Grove, PA) (goat anti-rabbit) was used at a dilution of 1:4000 and incubated for 1 h at room temperature. As a positive control, we used an anti-bactin antibody, 17 as previously described. 9,10 Proteins were revealed using a Western Lightning Plus-ECL Kit (PerkinElmer, Waltham, MA).…”

Section: Rt-pcrmentioning

confidence: 99%

Lentiviral transfer of an inducible transgene expressing a soluble form of Gas1 causes glioma cell arrest, apoptosis and inhibits tumor growth

et al. 2010

View full text Add to dashboard Cite

Gliomas are the most frequent primary tumors of the central nervous system, and their clinical prognosis remains very poor. Because of the characteristics of gliomas, gene therapy appears as a potentially relevant strategy for their treatment. However, the inability of viral vectors to transfer the therapeutic genes to a significantly high number of tumor cells, due to their limited diffusion and distribution, remains a critical obstacle for their application treating gliomas. We have demonstrated that the overexpression of growth arrest specific1 (Gas1) induces cell arrest and apoptosis and eliminates glioma cells in vitro and when implanted in mice. To improve the therapeutic range of Gas1, we generated lentiviral vectors coding for a soluble form of Gas1. Here, we show that cells infected with this virus produce the mutant protein, that acting both in autocrine and paracrine manners, causes death of infected and neighbor cells, thus importantly enhancing the effect of Gas1. Furthermore, the administration of this vector, or cells expressing it, inhibit the growth of tumors inoculated in mice. We present a gene therapy strategy that increases the effect of the therapeutic molecule by eliminating not just the infected cells that express Gas1, but neighbor non-infected cells.

show abstract

Biochemical and histochemical analysis of 71 kDa dystrophin isoform (Dp71f) in rat brain

Cited by 51 publications

References 64 publications

Immunofluorescence mapping of dystrophin in the rat brain: astrocytes contain the splice variant Dp71f, but this is confined to subpopulations

Immunofluorescence mapping of dystrophin in the rat brain: astrocytes contain the splice variant Dp71f, but this is confined to subpopulations

Targeted-simultaneous expression of Gas1 and p53 using a bicistronic adenoviral vector in gliomas

Lentiviral transfer of an inducible transgene expressing a soluble form of Gas1 causes glioma cell arrest, apoptosis and inhibits tumor growth

Contact Info

Product

Resources

About