Limits...
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.

Show MeSH

Related in: MedlinePlus

Cholera toxin regulates Hes3 localization in NSCs.(a, b) CholAB (2-d treatment) promotes the nuclear localization of Hes3 in the presence (6 days in FGF2) and absence of FGF2 in fetal NSC cultures. [Size bars: 20 µm].
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2877108&req=5

pone-0010841-g003: Cholera toxin regulates Hes3 localization in NSCs.(a, b) CholAB (2-d treatment) promotes the nuclear localization of Hes3 in the presence (6 days in FGF2) and absence of FGF2 in fetal NSC cultures. [Size bars: 20 µm].

Mentions: Downstream of Tie2 signaling is a pathway that induces Hes3 expression and regulates NSC survival [23]. In culture conditions that support the self-renewal of mouse fetal NSCs in vitro (in the presence of FGF2), Hes3 is localized both in the nucleus and the cytoplasm of fetal NSCs (Figure 3a,b; Figure S2; Figure S3), whereas when differentiation is induced (by a 2-day FGF2 withdrawal), Hes3 is only found in the cytoplasm. (Eventually, after approximately a total of 7 days of differentiation, all Hes3 signal is lost [25]). This suggests that nuclear Hes3 identifies the self-renewing, dividing NSC. When fetal NSCs were treated with CholAB for 2 days (following 4 days in FGF2), the nuclear localization of Hes3 markedly increased. This was true both in the presence and the absence of FGF2, suggesting that CholAB may promote the self-renewal state of these cultures and possibly maintain them in the division cycle even after mitogen (FGF2) withdrawal. We note that at earlier times within each passage (i.e. when the cultured cells are proliferating fast) a greater percentage of nuclear Hes3 is observed than at later times as used here (i.e. when the cultures are reaching confluence and the proliferation rates decrease) (data not shown).


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 Hes3 localization in NSCs.(a, b) CholAB (2-d treatment) promotes the nuclear localization of Hes3 in the presence (6 days in FGF2) and absence of FGF2 in fetal NSC cultures. [Size bars: 20 µm].
© Copyright Policy
Related In: Results  -  Collection

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

pone-0010841-g003: Cholera toxin regulates Hes3 localization in NSCs.(a, b) CholAB (2-d treatment) promotes the nuclear localization of Hes3 in the presence (6 days in FGF2) and absence of FGF2 in fetal NSC cultures. [Size bars: 20 µm].
Mentions: Downstream of Tie2 signaling is a pathway that induces Hes3 expression and regulates NSC survival [23]. In culture conditions that support the self-renewal of mouse fetal NSCs in vitro (in the presence of FGF2), Hes3 is localized both in the nucleus and the cytoplasm of fetal NSCs (Figure 3a,b; Figure S2; Figure S3), whereas when differentiation is induced (by a 2-day FGF2 withdrawal), Hes3 is only found in the cytoplasm. (Eventually, after approximately a total of 7 days of differentiation, all Hes3 signal is lost [25]). This suggests that nuclear Hes3 identifies the self-renewing, dividing NSC. When fetal NSCs were treated with CholAB for 2 days (following 4 days in FGF2), the nuclear localization of Hes3 markedly increased. This was true both in the presence and the absence of FGF2, suggesting that CholAB may promote the self-renewal state of these cultures and possibly maintain them in the division cycle even after mitogen (FGF2) withdrawal. We note that at earlier times within each passage (i.e. when the cultured cells are proliferating fast) a greater percentage of nuclear Hes3 is observed than at later times as used here (i.e. when the cultures are reaching confluence and the proliferation rates decrease) (data not shown).

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