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Cholera toxin regulates a signaling pathway critical for the expansion of neural stem cell cultures from the fetal and adult rodent brains.

Androutsellis-Theotokis A, Walbridge S, Park DM, Lonser RR, McKay RD - PLoS ONE (2010)

Bottom Line: Here we show that Cholera toxin regulates two recently characterized NSC markers, the Tie2 receptor and the transcription factor Hes3, and promotes the expansion of NSCs in culture.Cholera toxin increases immunoreactivity for the Tie2 receptor and rapidly induces the nuclear localization of Hes3.This is followed by powerful cultured NSC expansion and induction of proliferation both in the presence and absence of mitogen.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Biology, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland, USA. AndreasTheotokis@gmail.com

ABSTRACT

Background: New mechanisms that regulate neural stem cell (NSC) expansion will contribute to improved assay systems and the emerging regenerative approach that targets endogenous stem cells. Expanding knowledge on the control of stem cell self renewal will also lead to new approaches for targeting the stem cell population of cancers.

Methodology/principal findings: Here we show that Cholera toxin regulates two recently characterized NSC markers, the Tie2 receptor and the transcription factor Hes3, and promotes the expansion of NSCs in culture. Cholera toxin increases immunoreactivity for the Tie2 receptor and rapidly induces the nuclear localization of Hes3. This is followed by powerful cultured NSC expansion and induction of proliferation both in the presence and absence of mitogen.

Conclusions/significance: Our data suggest a new cell biological mechanism that regulates the self renewal and differentiation properties of stem cells, providing a new logic to manipulate NSCs in the context of regenerative disease and cancer.

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Related in: MedlinePlus

Cholera toxin regulates Tie2 receptors on NSCs.(a, b) High power magnifications of Tie2+ intracellular structures in cultured fetal NSCs. (c) CholAB (2-d treatment) increases the occurrence of Tie2+ structures in adult NSC cultures. [Size bars: 20 µm].
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pone-0010841-g002: Cholera toxin regulates Tie2 receptors on NSCs.(a, b) High power magnifications of Tie2+ intracellular structures in cultured fetal NSCs. (c) CholAB (2-d treatment) increases the occurrence of Tie2+ structures in adult NSC cultures. [Size bars: 20 µm].

Mentions: A recent paper cautions that immunocytochemical measurement of ganglioside expression may overestimate their expression (which can be independently determined by biochemical means) [31]. Here we have scored ganglioside expressing cells using a fluorescence conjugated cholera B subunit product (Invitrogen) and high-power confocal microscopy. Cells exhibiting labeling with the characteristic shape of lipid rafts were considered positive, thus avoiding false positives from background signal. We do note, however, that the signal-to-noise ratio was very high as can be seen in Figure 2a and Figure S2. Consistent with this previous study, we find that cultured NSCs exhibit low levels of Cholera toxin B subunit binding, but we also do caution here of the need to perform ganglioside expression studies carefully.


Cholera toxin regulates a signaling pathway critical for the expansion of neural stem cell cultures from the fetal and adult rodent brains.

Androutsellis-Theotokis A, Walbridge S, Park DM, Lonser RR, McKay RD - PLoS ONE (2010)

Cholera toxin regulates Tie2 receptors on NSCs.(a, b) High power magnifications of Tie2+ intracellular structures in cultured fetal NSCs. (c) CholAB (2-d treatment) increases the occurrence of Tie2+ structures in adult NSC cultures. [Size bars: 20 µm].
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2877108&req=5

pone-0010841-g002: Cholera toxin regulates Tie2 receptors on NSCs.(a, b) High power magnifications of Tie2+ intracellular structures in cultured fetal NSCs. (c) CholAB (2-d treatment) increases the occurrence of Tie2+ structures in adult NSC cultures. [Size bars: 20 µm].
Mentions: A recent paper cautions that immunocytochemical measurement of ganglioside expression may overestimate their expression (which can be independently determined by biochemical means) [31]. Here we have scored ganglioside expressing cells using a fluorescence conjugated cholera B subunit product (Invitrogen) and high-power confocal microscopy. Cells exhibiting labeling with the characteristic shape of lipid rafts were considered positive, thus avoiding false positives from background signal. We do note, however, that the signal-to-noise ratio was very high as can be seen in Figure 2a and Figure S2. Consistent with this previous study, we find that cultured NSCs exhibit low levels of Cholera toxin B subunit binding, but we also do caution here of the need to perform ganglioside expression studies carefully.

Bottom Line: Here we show that Cholera toxin regulates two recently characterized NSC markers, the Tie2 receptor and the transcription factor Hes3, and promotes the expansion of NSCs in culture.Cholera toxin increases immunoreactivity for the Tie2 receptor and rapidly induces the nuclear localization of Hes3.This is followed by powerful cultured NSC expansion and induction of proliferation both in the presence and absence of mitogen.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Biology, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland, USA. AndreasTheotokis@gmail.com

ABSTRACT

Background: New mechanisms that regulate neural stem cell (NSC) expansion will contribute to improved assay systems and the emerging regenerative approach that targets endogenous stem cells. Expanding knowledge on the control of stem cell self renewal will also lead to new approaches for targeting the stem cell population of cancers.

Methodology/principal findings: Here we show that Cholera toxin regulates two recently characterized NSC markers, the Tie2 receptor and the transcription factor Hes3, and promotes the expansion of NSCs in culture. Cholera toxin increases immunoreactivity for the Tie2 receptor and rapidly induces the nuclear localization of Hes3. This is followed by powerful cultured NSC expansion and induction of proliferation both in the presence and absence of mitogen.

Conclusions/significance: Our data suggest a new cell biological mechanism that regulates the self renewal and differentiation properties of stem cells, providing a new logic to manipulate NSCs in the context of regenerative disease and cancer.

Show MeSH
Related in: MedlinePlus