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Neurons produce FGF2 and VEGF and secrete them at least in part by shedding extracellular vesicles.

Schiera G, Proia P, Alberti C, Mineo M, Savettieri G, Di Liegro I - J. Cell. Mol. Med. (2007 Nov-Dec)

Bottom Line: We previously found that neurons are able to affect the ability of brain capillary endothelial cells to form in vitro a monolayer with properties resembling the blood-brain barrier.In the present paper, we report that neurons produce both vascular endothelial growth factor and fibroblast growth factor 2, two well-known angiogenic factors.Shedding of extracellular vesicles by neurons was also confirmed by scanner electron microscopy.

View Article: PubMed Central - PubMed

Affiliation: Dipartimento di Scienze Biochimiche, Università degli Studi di Palermo, Palermo, Italy.

ABSTRACT
We previously found that neurons are able to affect the ability of brain capillary endothelial cells to form in vitro a monolayer with properties resembling the blood-brain barrier. We then looked, by immunofluorescence and western analysis, for factors, produced by neurons, with the potential to influence growth and differentiation of endothelial cells. In the present paper, we report that neurons produce both vascular endothelial growth factor and fibroblast growth factor 2, two well-known angiogenic factors. More interestingly, we gained evidence that both factors are released by neurons, at least in part, by shedding of extracellular vesicles, that contain beta1 integrin, a membrane protein already known to be part of extracellular vesicles released by tumour cells. Shedding of extracellular vesicles by neurons was also confirmed by scanner electron microscopy.

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Colocalization of either FGF-2 or VEGF with β1 integrin in extracellular vesicles. Double immunofluorescence was used to co-localize either FGF-2 (b, b*: red) or VEGF (e, e*: red) and β1-integrin (a, a*; d, d*: green) immunoreactivity in neurons cultured for 15 days in Maat Medium. Overlay of FGF-2 and b1-integrin fluorescence is shown in c and c*. Overlay of VEGF and β1-integrin fluorescence is shown in f and f*. a*–f*, enlarged views of the fields boxed in the figures a–f. Cells were observed in an Olympus fluoview FV300 confocal microscope, equipped with two laser multipliers.
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fig03: Colocalization of either FGF-2 or VEGF with β1 integrin in extracellular vesicles. Double immunofluorescence was used to co-localize either FGF-2 (b, b*: red) or VEGF (e, e*: red) and β1-integrin (a, a*; d, d*: green) immunoreactivity in neurons cultured for 15 days in Maat Medium. Overlay of FGF-2 and b1-integrin fluorescence is shown in c and c*. Overlay of VEGF and β1-integrin fluorescence is shown in f and f*. a*–f*, enlarged views of the fields boxed in the figures a–f. Cells were observed in an Olympus fluoview FV300 confocal microscope, equipped with two laser multipliers.

Mentions: MVs have been reported to originate from domains of the plasma membrane selectively enriched in specific membrane components, such as β1 integrin and membrane-bound gelatinase B/MMP-9 [13]. Therefore we investigated, by double immunofluorescence and confocal microscopy, the possible colocalization of both FGF-2 and VEGF with β1 integrin. The results confirmed, first of all, that neurons produce both FGF-2 and VEGF. As shown in Fig. 3, indeed, neurons were immunostained by both anti-FGF-2 antibodies (b, b*) and anti-VEGF antibodies (e, e*). Second, we found that both molecules are at least in part present in round extracellular structures, emanating from neurons, also stained by anti-β1 integrin antibodies (a, a*; d, d*). The sizes of the extracellular structures, visible in Fig. 3, are comparable with the sizes of those visible in Figs. 1 and 2. We noticed in some cases a tendency of vesicles to cluster around neurites (see, e.g.Fig. 3f*, structures indicated by arrows).


Neurons produce FGF2 and VEGF and secrete them at least in part by shedding extracellular vesicles.

Schiera G, Proia P, Alberti C, Mineo M, Savettieri G, Di Liegro I - J. Cell. Mol. Med. (2007 Nov-Dec)

Colocalization of either FGF-2 or VEGF with β1 integrin in extracellular vesicles. Double immunofluorescence was used to co-localize either FGF-2 (b, b*: red) or VEGF (e, e*: red) and β1-integrin (a, a*; d, d*: green) immunoreactivity in neurons cultured for 15 days in Maat Medium. Overlay of FGF-2 and b1-integrin fluorescence is shown in c and c*. Overlay of VEGF and β1-integrin fluorescence is shown in f and f*. a*–f*, enlarged views of the fields boxed in the figures a–f. Cells were observed in an Olympus fluoview FV300 confocal microscope, equipped with two laser multipliers.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4401300&req=5

fig03: Colocalization of either FGF-2 or VEGF with β1 integrin in extracellular vesicles. Double immunofluorescence was used to co-localize either FGF-2 (b, b*: red) or VEGF (e, e*: red) and β1-integrin (a, a*; d, d*: green) immunoreactivity in neurons cultured for 15 days in Maat Medium. Overlay of FGF-2 and b1-integrin fluorescence is shown in c and c*. Overlay of VEGF and β1-integrin fluorescence is shown in f and f*. a*–f*, enlarged views of the fields boxed in the figures a–f. Cells were observed in an Olympus fluoview FV300 confocal microscope, equipped with two laser multipliers.
Mentions: MVs have been reported to originate from domains of the plasma membrane selectively enriched in specific membrane components, such as β1 integrin and membrane-bound gelatinase B/MMP-9 [13]. Therefore we investigated, by double immunofluorescence and confocal microscopy, the possible colocalization of both FGF-2 and VEGF with β1 integrin. The results confirmed, first of all, that neurons produce both FGF-2 and VEGF. As shown in Fig. 3, indeed, neurons were immunostained by both anti-FGF-2 antibodies (b, b*) and anti-VEGF antibodies (e, e*). Second, we found that both molecules are at least in part present in round extracellular structures, emanating from neurons, also stained by anti-β1 integrin antibodies (a, a*; d, d*). The sizes of the extracellular structures, visible in Fig. 3, are comparable with the sizes of those visible in Figs. 1 and 2. We noticed in some cases a tendency of vesicles to cluster around neurites (see, e.g.Fig. 3f*, structures indicated by arrows).

Bottom Line: We previously found that neurons are able to affect the ability of brain capillary endothelial cells to form in vitro a monolayer with properties resembling the blood-brain barrier.In the present paper, we report that neurons produce both vascular endothelial growth factor and fibroblast growth factor 2, two well-known angiogenic factors.Shedding of extracellular vesicles by neurons was also confirmed by scanner electron microscopy.

View Article: PubMed Central - PubMed

Affiliation: Dipartimento di Scienze Biochimiche, Università degli Studi di Palermo, Palermo, Italy.

ABSTRACT
We previously found that neurons are able to affect the ability of brain capillary endothelial cells to form in vitro a monolayer with properties resembling the blood-brain barrier. We then looked, by immunofluorescence and western analysis, for factors, produced by neurons, with the potential to influence growth and differentiation of endothelial cells. In the present paper, we report that neurons produce both vascular endothelial growth factor and fibroblast growth factor 2, two well-known angiogenic factors. More interestingly, we gained evidence that both factors are released by neurons, at least in part, by shedding of extracellular vesicles, that contain beta1 integrin, a membrane protein already known to be part of extracellular vesicles released by tumour cells. Shedding of extracellular vesicles by neurons was also confirmed by scanner electron microscopy.

Show MeSH
Related in: MedlinePlus