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Postnatal NG2 proteoglycan-expressing progenitor cells are intrinsically multipotent and generate functional neurons.

Belachew S, Chittajallu R, Aguirre AA, Yuan X, Kirby M, Anderson S, Gallo V - J. Cell Biol. (2003)

Bottom Line: The fast kinetics and the high rate of multipotent fate of these NG2+ progenitors in vitro reflect an intrinsic property, rather than reprogramming.We demonstrate in the hippocampus in vivo that a sizeable fraction of postnatal NG2+ progenitor cells are proliferative precursors whose progeny appears to differentiate into GABAergic neurons capable of propagating action potentials and displaying functional synaptic inputs.These data show that at least a subpopulation of postnatal NG2-expressing cells are CNS multipotent precursors that may underlie adult hippocampal neurogenesis.

View Article: PubMed Central - PubMed

Affiliation: Center for Neuroscience Research, Children's Research Institute, Children's National Medical Center, Washington, DC 20010-2970, USA.

ABSTRACT
Neurogenesis is known to persist in the adult mammalian central nervous system (CNS). The identity of the cells that generate new neurons in the postnatal CNS has become a crucial but elusive issue. Using a transgenic mouse, we show that NG2 proteoglycan-positive progenitor cells that express the 2',3'-cyclic nucleotide 3'-phosphodiesterase gene display a multipotent phenotype in vitro and generate electrically excitable neurons, as well as astrocytes and oligodendrocytes. The fast kinetics and the high rate of multipotent fate of these NG2+ progenitors in vitro reflect an intrinsic property, rather than reprogramming. We demonstrate in the hippocampus in vivo that a sizeable fraction of postnatal NG2+ progenitor cells are proliferative precursors whose progeny appears to differentiate into GABAergic neurons capable of propagating action potentials and displaying functional synaptic inputs. These data show that at least a subpopulation of postnatal NG2-expressing cells are CNS multipotent precursors that may underlie adult hippocampal neurogenesis.

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Early postnatal CNP-GFP+ cells are proliferative and display an NG2+/nestin+ phenotype in vivo in the SVZ and hippocampus. As illustrated by Z-series (A–D, 22-μm thickness, image steps = 0.5 μm) confocal scanning images of the same field in SVZ area from P2 CNP-GFP transgenic mice, CNP-GFP+ cells expressed an NG2+/nestin+ phenotype. 1-μm thick single plane images of a single cell at high magnification are provided in insets of A–D. The same phenotype was observed in the hippocampus (not depicted). (A) GFP green fluorescence, (B) NG2 staining, (C) nestin staining, (D) overlay. As shown in E–J (high magnification in insets), CNP-GFP+ cells were proliferative in vivo. 0.5-μm thick single plane confocal scanning images of CA3 stratum radiatum area of hippocampus (E–G) and SVZ (H–J) from P2 CNP-GFP transgenic mice. (E and H) GFP green fluorescence, (F and I) PCNA staining, (G and J) overlay. A high percentage of CNP-GFP+ cells were PCNA+ in both germinative areas. Bars: 50 μm (A–D) and (E–J).
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fig5: Early postnatal CNP-GFP+ cells are proliferative and display an NG2+/nestin+ phenotype in vivo in the SVZ and hippocampus. As illustrated by Z-series (A–D, 22-μm thickness, image steps = 0.5 μm) confocal scanning images of the same field in SVZ area from P2 CNP-GFP transgenic mice, CNP-GFP+ cells expressed an NG2+/nestin+ phenotype. 1-μm thick single plane images of a single cell at high magnification are provided in insets of A–D. The same phenotype was observed in the hippocampus (not depicted). (A) GFP green fluorescence, (B) NG2 staining, (C) nestin staining, (D) overlay. As shown in E–J (high magnification in insets), CNP-GFP+ cells were proliferative in vivo. 0.5-μm thick single plane confocal scanning images of CA3 stratum radiatum area of hippocampus (E–G) and SVZ (H–J) from P2 CNP-GFP transgenic mice. (E and H) GFP green fluorescence, (F and I) PCNA staining, (G and J) overlay. A high percentage of CNP-GFP+ cells were PCNA+ in both germinative areas. Bars: 50 μm (A–D) and (E–J).

Mentions: We performed an analysis of the properties of postnatal CNP-GFP+ cells in situ, with a particular focus on postnatal germinative regions (i.e., SVZ and hippocampus). Consistent with our data on acutely isolated CNP-GFP+ cells from the same developmental stage (Fig. 1 D), a high proportion of CNP-GFP+ cells were also NG2+ and nestin+ at P2 in situ (Fig. 5 , A–D). Direct cell counting demonstrated that 89.6 ± 1.7% (mean ± SEM, total cells counted = 483) of CNP-GFP+ cells were also NG2+ in the SVZ at P2, and 18.4 ± 1.6% (mean ± SEM, total cells counted = 273) of CNP-GFP+ cells were NG2+ in the hippocampus at P2. Because nestin immunostaining was mostly visualized in the cell processes (Fig. 5 C), we were unable to obtain an accurate quantification of the nestin phenotype in situ. Given the controversy over whether stem cells of the adult CNS are located in the ependymal or subependymal layer of lateral ventricle walls (Chiasson et al., 1999; Doetsch et al., 1999; Johansson et al., 1999; Laywell et al., 2000; Capela and Temple, 2002), it is important to point out that NG2+/CNP-GFP+ cells were found in the subependymal zone (i.e., SVZ) but never in the ependymal layer, from P2 to P30 (unpublished data).


Postnatal NG2 proteoglycan-expressing progenitor cells are intrinsically multipotent and generate functional neurons.

Belachew S, Chittajallu R, Aguirre AA, Yuan X, Kirby M, Anderson S, Gallo V - J. Cell Biol. (2003)

Early postnatal CNP-GFP+ cells are proliferative and display an NG2+/nestin+ phenotype in vivo in the SVZ and hippocampus. As illustrated by Z-series (A–D, 22-μm thickness, image steps = 0.5 μm) confocal scanning images of the same field in SVZ area from P2 CNP-GFP transgenic mice, CNP-GFP+ cells expressed an NG2+/nestin+ phenotype. 1-μm thick single plane images of a single cell at high magnification are provided in insets of A–D. The same phenotype was observed in the hippocampus (not depicted). (A) GFP green fluorescence, (B) NG2 staining, (C) nestin staining, (D) overlay. As shown in E–J (high magnification in insets), CNP-GFP+ cells were proliferative in vivo. 0.5-μm thick single plane confocal scanning images of CA3 stratum radiatum area of hippocampus (E–G) and SVZ (H–J) from P2 CNP-GFP transgenic mice. (E and H) GFP green fluorescence, (F and I) PCNA staining, (G and J) overlay. A high percentage of CNP-GFP+ cells were PCNA+ in both germinative areas. Bars: 50 μm (A–D) and (E–J).
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Related In: Results  -  Collection

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fig5: Early postnatal CNP-GFP+ cells are proliferative and display an NG2+/nestin+ phenotype in vivo in the SVZ and hippocampus. As illustrated by Z-series (A–D, 22-μm thickness, image steps = 0.5 μm) confocal scanning images of the same field in SVZ area from P2 CNP-GFP transgenic mice, CNP-GFP+ cells expressed an NG2+/nestin+ phenotype. 1-μm thick single plane images of a single cell at high magnification are provided in insets of A–D. The same phenotype was observed in the hippocampus (not depicted). (A) GFP green fluorescence, (B) NG2 staining, (C) nestin staining, (D) overlay. As shown in E–J (high magnification in insets), CNP-GFP+ cells were proliferative in vivo. 0.5-μm thick single plane confocal scanning images of CA3 stratum radiatum area of hippocampus (E–G) and SVZ (H–J) from P2 CNP-GFP transgenic mice. (E and H) GFP green fluorescence, (F and I) PCNA staining, (G and J) overlay. A high percentage of CNP-GFP+ cells were PCNA+ in both germinative areas. Bars: 50 μm (A–D) and (E–J).
Mentions: We performed an analysis of the properties of postnatal CNP-GFP+ cells in situ, with a particular focus on postnatal germinative regions (i.e., SVZ and hippocampus). Consistent with our data on acutely isolated CNP-GFP+ cells from the same developmental stage (Fig. 1 D), a high proportion of CNP-GFP+ cells were also NG2+ and nestin+ at P2 in situ (Fig. 5 , A–D). Direct cell counting demonstrated that 89.6 ± 1.7% (mean ± SEM, total cells counted = 483) of CNP-GFP+ cells were also NG2+ in the SVZ at P2, and 18.4 ± 1.6% (mean ± SEM, total cells counted = 273) of CNP-GFP+ cells were NG2+ in the hippocampus at P2. Because nestin immunostaining was mostly visualized in the cell processes (Fig. 5 C), we were unable to obtain an accurate quantification of the nestin phenotype in situ. Given the controversy over whether stem cells of the adult CNS are located in the ependymal or subependymal layer of lateral ventricle walls (Chiasson et al., 1999; Doetsch et al., 1999; Johansson et al., 1999; Laywell et al., 2000; Capela and Temple, 2002), it is important to point out that NG2+/CNP-GFP+ cells were found in the subependymal zone (i.e., SVZ) but never in the ependymal layer, from P2 to P30 (unpublished data).

Bottom Line: The fast kinetics and the high rate of multipotent fate of these NG2+ progenitors in vitro reflect an intrinsic property, rather than reprogramming.We demonstrate in the hippocampus in vivo that a sizeable fraction of postnatal NG2+ progenitor cells are proliferative precursors whose progeny appears to differentiate into GABAergic neurons capable of propagating action potentials and displaying functional synaptic inputs.These data show that at least a subpopulation of postnatal NG2-expressing cells are CNS multipotent precursors that may underlie adult hippocampal neurogenesis.

View Article: PubMed Central - PubMed

Affiliation: Center for Neuroscience Research, Children's Research Institute, Children's National Medical Center, Washington, DC 20010-2970, USA.

ABSTRACT
Neurogenesis is known to persist in the adult mammalian central nervous system (CNS). The identity of the cells that generate new neurons in the postnatal CNS has become a crucial but elusive issue. Using a transgenic mouse, we show that NG2 proteoglycan-positive progenitor cells that express the 2',3'-cyclic nucleotide 3'-phosphodiesterase gene display a multipotent phenotype in vitro and generate electrically excitable neurons, as well as astrocytes and oligodendrocytes. The fast kinetics and the high rate of multipotent fate of these NG2+ progenitors in vitro reflect an intrinsic property, rather than reprogramming. We demonstrate in the hippocampus in vivo that a sizeable fraction of postnatal NG2+ progenitor cells are proliferative precursors whose progeny appears to differentiate into GABAergic neurons capable of propagating action potentials and displaying functional synaptic inputs. These data show that at least a subpopulation of postnatal NG2-expressing cells are CNS multipotent precursors that may underlie adult hippocampal neurogenesis.

Show MeSH
Related in: MedlinePlus