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ApoTransferrin: dual role on adult subventricular zone-derived neurospheres.

Silvestroff L, Franco PG, Pasquini JM - PLoS ONE (2012)

Bottom Line: We cultured NSC/NPCs from the rat subventricular zone as neurospheres (NS) and studied apoTransferrin (aTf) effects on oligodendroglial specification and maturation.On the other hand, aTf exposure during differentiating conditions favours OL maturation from OPCs by promoting OL morphological development.This evidence supports a key role of Tf on the generation of OL from NSC/NPCs and highlights its potential in demyelinating disorder treatment.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Química Biológica, Instituto de Química y Fisicoquímica Biológica, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina.

ABSTRACT
Neural stem and progenitor cells (NSC/NPCs) are multipotent self-renewing cells that are able to generate neurons, astrocytes and oligodendrocytes (OLs) within the adult central nervous system. We cultured NSC/NPCs from the rat subventricular zone as neurospheres (NS) and studied apoTransferrin (aTf) effects on oligodendroglial specification and maturation. Our findings suggest that aTf acts at different stages during progression from NSC to mature oligodendrocytes. On the one hand, an early event associated with the activation of NSC/NPCs proliferation and commitment toward the oligodendroglial fate, as indicated by increased BrdU incorporation, larger neurospheres production, and higher ability to generate OL precursors (OPCs) from undifferentiated cultures. On the other hand, aTf exposure during differentiating conditions favours OL maturation from OPCs by promoting OL morphological development. This evidence supports a key role of Tf on the generation of OL from NSC/NPCs and highlights its potential in demyelinating disorder treatment.

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Transferrin Receptor (TfRc) expression.TfRc was immunodetected with the CD71 monoclonal antibody. During proliferation, NG2+ cells (A, green) and PDGFRα+ cells (B, green) co-express the TfRc (red). Empty arrowheads in A indicate NG2+/CD71+ cells. Filled arrowheads indicate bipolar-shaped NG2−/CD71+. After differentiation, GFAP+ (C, green) cells were found expressing TfRc (red). MBP+ cells (D, green) show strong CD71 immunolabelling (red). Confocal image analysis reinforces the concept of MBP+ OL expressing the TfRc (E–H). The blue colour in images indicates Höechst nuclear dye. Scale bar in A equals 100 µm in A and D, scale bar in C equals 50 µm, and scale bar in E equals 40 µm for E–H.
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pone-0033937-g005: Transferrin Receptor (TfRc) expression.TfRc was immunodetected with the CD71 monoclonal antibody. During proliferation, NG2+ cells (A, green) and PDGFRα+ cells (B, green) co-express the TfRc (red). Empty arrowheads in A indicate NG2+/CD71+ cells. Filled arrowheads indicate bipolar-shaped NG2−/CD71+. After differentiation, GFAP+ (C, green) cells were found expressing TfRc (red). MBP+ cells (D, green) show strong CD71 immunolabelling (red). Confocal image analysis reinforces the concept of MBP+ OL expressing the TfRc (E–H). The blue colour in images indicates Höechst nuclear dye. Scale bar in A equals 100 µm in A and D, scale bar in C equals 50 µm, and scale bar in E equals 40 µm for E–H.

Mentions: Since the canonical mechanism of cellular uptake of Tf is mediated by Tf Receptor (TfRc), we evaluated the presence of TfRc in dissociated NS cells at different time points during culture progression by immunocytochemistry with the anti CD71 monoclonal antibody. These descriptive results confirm that the TfRc was detected at low or moderate levels in all cell types in vitro. At the end of the proliferation condition CD71+ cells co-localized with the progenitor marker NG2 (Fig. 5A, empty arrowheads) and the OPC marker PDGFRα (Fig. 5B). TfRc expression in these cells was restricted to a peri-nuclear region in the cell soma. Other CD71+ cells were also detected which did not express either NG2 nor PDGFRα, and had a bipolar morphology (whole arrowheads in Fig. 5A). After differentiation, GFAP+ cells expressed low TfRc levels (Fig. 5C), and intermediate levels were found in MBP+ OLs (Fig. 5D). We used confocal images to show TfRc immunostaining colocalized with MBP+ cells, and that the former was not only located on the cell body surface, but expressed on process membranes as well (Fig. 5E–H). In order to determine the presence of TfRc in the SVZ tissue, we used SVZ explants cultures to avoid the undesired CD71 high expression normally seen in vasculature in vivo. Although tissue explants are grown in vitro, most of the cytoarchitecture remains unchanged, rendering the cellular environment somewhat similar to the one found in the original tissue. In these cultures, TfRc immunoreactivity was mainly found in cells within the explants itself (Supp Fig S4A), while Tf+ cells were found on cells that had migrated away from the explant (Supp Fig S4B), where a few cells displayed simultaneous Tf and TfRc immunoreactivity (Supp Fig S4C, C′). Thus, Tf signalling is actually present in the cells derived from the lateral ventricle wall, which could effectively be functional Tf cell targets.


ApoTransferrin: dual role on adult subventricular zone-derived neurospheres.

Silvestroff L, Franco PG, Pasquini JM - PLoS ONE (2012)

Transferrin Receptor (TfRc) expression.TfRc was immunodetected with the CD71 monoclonal antibody. During proliferation, NG2+ cells (A, green) and PDGFRα+ cells (B, green) co-express the TfRc (red). Empty arrowheads in A indicate NG2+/CD71+ cells. Filled arrowheads indicate bipolar-shaped NG2−/CD71+. After differentiation, GFAP+ (C, green) cells were found expressing TfRc (red). MBP+ cells (D, green) show strong CD71 immunolabelling (red). Confocal image analysis reinforces the concept of MBP+ OL expressing the TfRc (E–H). The blue colour in images indicates Höechst nuclear dye. Scale bar in A equals 100 µm in A and D, scale bar in C equals 50 µm, and scale bar in E equals 40 µm for E–H.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3316520&req=5

pone-0033937-g005: Transferrin Receptor (TfRc) expression.TfRc was immunodetected with the CD71 monoclonal antibody. During proliferation, NG2+ cells (A, green) and PDGFRα+ cells (B, green) co-express the TfRc (red). Empty arrowheads in A indicate NG2+/CD71+ cells. Filled arrowheads indicate bipolar-shaped NG2−/CD71+. After differentiation, GFAP+ (C, green) cells were found expressing TfRc (red). MBP+ cells (D, green) show strong CD71 immunolabelling (red). Confocal image analysis reinforces the concept of MBP+ OL expressing the TfRc (E–H). The blue colour in images indicates Höechst nuclear dye. Scale bar in A equals 100 µm in A and D, scale bar in C equals 50 µm, and scale bar in E equals 40 µm for E–H.
Mentions: Since the canonical mechanism of cellular uptake of Tf is mediated by Tf Receptor (TfRc), we evaluated the presence of TfRc in dissociated NS cells at different time points during culture progression by immunocytochemistry with the anti CD71 monoclonal antibody. These descriptive results confirm that the TfRc was detected at low or moderate levels in all cell types in vitro. At the end of the proliferation condition CD71+ cells co-localized with the progenitor marker NG2 (Fig. 5A, empty arrowheads) and the OPC marker PDGFRα (Fig. 5B). TfRc expression in these cells was restricted to a peri-nuclear region in the cell soma. Other CD71+ cells were also detected which did not express either NG2 nor PDGFRα, and had a bipolar morphology (whole arrowheads in Fig. 5A). After differentiation, GFAP+ cells expressed low TfRc levels (Fig. 5C), and intermediate levels were found in MBP+ OLs (Fig. 5D). We used confocal images to show TfRc immunostaining colocalized with MBP+ cells, and that the former was not only located on the cell body surface, but expressed on process membranes as well (Fig. 5E–H). In order to determine the presence of TfRc in the SVZ tissue, we used SVZ explants cultures to avoid the undesired CD71 high expression normally seen in vasculature in vivo. Although tissue explants are grown in vitro, most of the cytoarchitecture remains unchanged, rendering the cellular environment somewhat similar to the one found in the original tissue. In these cultures, TfRc immunoreactivity was mainly found in cells within the explants itself (Supp Fig S4A), while Tf+ cells were found on cells that had migrated away from the explant (Supp Fig S4B), where a few cells displayed simultaneous Tf and TfRc immunoreactivity (Supp Fig S4C, C′). Thus, Tf signalling is actually present in the cells derived from the lateral ventricle wall, which could effectively be functional Tf cell targets.

Bottom Line: We cultured NSC/NPCs from the rat subventricular zone as neurospheres (NS) and studied apoTransferrin (aTf) effects on oligodendroglial specification and maturation.On the other hand, aTf exposure during differentiating conditions favours OL maturation from OPCs by promoting OL morphological development.This evidence supports a key role of Tf on the generation of OL from NSC/NPCs and highlights its potential in demyelinating disorder treatment.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Química Biológica, Instituto de Química y Fisicoquímica Biológica, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina.

ABSTRACT
Neural stem and progenitor cells (NSC/NPCs) are multipotent self-renewing cells that are able to generate neurons, astrocytes and oligodendrocytes (OLs) within the adult central nervous system. We cultured NSC/NPCs from the rat subventricular zone as neurospheres (NS) and studied apoTransferrin (aTf) effects on oligodendroglial specification and maturation. Our findings suggest that aTf acts at different stages during progression from NSC to mature oligodendrocytes. On the one hand, an early event associated with the activation of NSC/NPCs proliferation and commitment toward the oligodendroglial fate, as indicated by increased BrdU incorporation, larger neurospheres production, and higher ability to generate OL precursors (OPCs) from undifferentiated cultures. On the other hand, aTf exposure during differentiating conditions favours OL maturation from OPCs by promoting OL morphological development. This evidence supports a key role of Tf on the generation of OL from NSC/NPCs and highlights its potential in demyelinating disorder treatment.

Show MeSH
Related in: MedlinePlus