Limits...
Hierarchical differentiation competence in response to retinoic acid ensures stem cell maintenance during mouse spermatogenesis.

Ikami K, Tokue M, Sugimoto R, Noda C, Kobayashi S, Hara K, Yoshida S - Development (2015)

Bottom Line: Ectopic expression of RARγ was sufficient to induce GFRα1(+) cells to directly differentiate to KIT(+) cells without transiting the NGN3(+) state.Therefore, RARγ plays key roles in the differentiation competence of NGN3(+) cells.We propose a novel mechanism of stem cell fate selection in an open niche environment whereby undifferentiated cells show heterogeneous competence to differentiate in response to ubiquitously distributed differentiation-inducing signals.

View Article: PubMed Central - PubMed

Affiliation: Division of Germ Cell Biology, National Institute for Basic Biology, National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji, Okazaki 444-8787, Japan Department of Basic Biology, School of Life Science, Graduate University for Advanced Studies (Sokendai), 5-1 Higashiyama, Myodaiji, Okazaki 444-8787, Japan.

Show MeSH

Related in: MedlinePlus

Fate of pulse-labeled NGN3+ spermatogonia following administration of VA. (A) Experimental design. Two days after the TM pulse, Ngn3-CreERTM; CAG-CAT-EGFP transgenic mice maintained in VAD were injected with VA to resume spermatogenesis. Testis samples were harvested at the indicated times. (B) IF analysis of GFP and KIT expression in whole-mount seminiferous tubules 0, 2, 4 and 6 days after VA injection. Arrowheads indicate GFP+/KIT+ double-positive cells. Note that KIT immunostaining exhibits a punctate pattern at these early stages. (C) The number of GFP-labeled GFRα1+ Aundiff (magenta), GFRα1− Aundiff (green), KIT+ (blue) and total (black) cells in the testes of Ngn3-CreERTM; CAG-CAT-EGFP mice. Shown is the mean±s.e.m. of three, three, five and five testes on days 0, 2, 4 and 6, respectively. *P=0.041, **P<0.002 (t-test) compared with the values for day 0. Scale bar: 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4419276&req=5

DEV118695F2: Fate of pulse-labeled NGN3+ spermatogonia following administration of VA. (A) Experimental design. Two days after the TM pulse, Ngn3-CreERTM; CAG-CAT-EGFP transgenic mice maintained in VAD were injected with VA to resume spermatogenesis. Testis samples were harvested at the indicated times. (B) IF analysis of GFP and KIT expression in whole-mount seminiferous tubules 0, 2, 4 and 6 days after VA injection. Arrowheads indicate GFP+/KIT+ double-positive cells. Note that KIT immunostaining exhibits a punctate pattern at these early stages. (C) The number of GFP-labeled GFRα1+ Aundiff (magenta), GFRα1− Aundiff (green), KIT+ (blue) and total (black) cells in the testes of Ngn3-CreERTM; CAG-CAT-EGFP mice. Shown is the mean±s.e.m. of three, three, five and five testes on days 0, 2, 4 and 6, respectively. *P=0.041, **P<0.002 (t-test) compared with the values for day 0. Scale bar: 50 μm.

Mentions: We first pulse-labeled the NGN3+ cells and traced their fates. NGN3+ spermatogonia were irreversibly labeled with green fluorescent protein (GFP) after a single pulse of 4-hydroxytamoxifen (TM) was administered to Ngn3-CreERTM; CAG-CAT-EGFP transgenic mice maintained under conditions of VAD. Following administration of VA, gene expression by the GFP-labeled cells was analyzed using whole-mount immunofluorescence (Fig. 2A). In VAD (day 0), almost all the labeled cells were negative for KIT and GFRα1 expression, as expected. After injecting VA, almost all the GFP-labeled cells expressed KIT within 4 days (Fig. 2B,C), whereas they remained GFRα1− throughout (Fig. 2C; supplementary material Fig. S1A,B). These data indicate that NGN3+ cells transit to KIT+ cells rapidly and efficiently in response to RA.Fig. 2.


Hierarchical differentiation competence in response to retinoic acid ensures stem cell maintenance during mouse spermatogenesis.

Ikami K, Tokue M, Sugimoto R, Noda C, Kobayashi S, Hara K, Yoshida S - Development (2015)

Fate of pulse-labeled NGN3+ spermatogonia following administration of VA. (A) Experimental design. Two days after the TM pulse, Ngn3-CreERTM; CAG-CAT-EGFP transgenic mice maintained in VAD were injected with VA to resume spermatogenesis. Testis samples were harvested at the indicated times. (B) IF analysis of GFP and KIT expression in whole-mount seminiferous tubules 0, 2, 4 and 6 days after VA injection. Arrowheads indicate GFP+/KIT+ double-positive cells. Note that KIT immunostaining exhibits a punctate pattern at these early stages. (C) The number of GFP-labeled GFRα1+ Aundiff (magenta), GFRα1− Aundiff (green), KIT+ (blue) and total (black) cells in the testes of Ngn3-CreERTM; CAG-CAT-EGFP mice. Shown is the mean±s.e.m. of three, three, five and five testes on days 0, 2, 4 and 6, respectively. *P=0.041, **P<0.002 (t-test) compared with the values for day 0. Scale bar: 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4419276&req=5

DEV118695F2: Fate of pulse-labeled NGN3+ spermatogonia following administration of VA. (A) Experimental design. Two days after the TM pulse, Ngn3-CreERTM; CAG-CAT-EGFP transgenic mice maintained in VAD were injected with VA to resume spermatogenesis. Testis samples were harvested at the indicated times. (B) IF analysis of GFP and KIT expression in whole-mount seminiferous tubules 0, 2, 4 and 6 days after VA injection. Arrowheads indicate GFP+/KIT+ double-positive cells. Note that KIT immunostaining exhibits a punctate pattern at these early stages. (C) The number of GFP-labeled GFRα1+ Aundiff (magenta), GFRα1− Aundiff (green), KIT+ (blue) and total (black) cells in the testes of Ngn3-CreERTM; CAG-CAT-EGFP mice. Shown is the mean±s.e.m. of three, three, five and five testes on days 0, 2, 4 and 6, respectively. *P=0.041, **P<0.002 (t-test) compared with the values for day 0. Scale bar: 50 μm.
Mentions: We first pulse-labeled the NGN3+ cells and traced their fates. NGN3+ spermatogonia were irreversibly labeled with green fluorescent protein (GFP) after a single pulse of 4-hydroxytamoxifen (TM) was administered to Ngn3-CreERTM; CAG-CAT-EGFP transgenic mice maintained under conditions of VAD. Following administration of VA, gene expression by the GFP-labeled cells was analyzed using whole-mount immunofluorescence (Fig. 2A). In VAD (day 0), almost all the labeled cells were negative for KIT and GFRα1 expression, as expected. After injecting VA, almost all the GFP-labeled cells expressed KIT within 4 days (Fig. 2B,C), whereas they remained GFRα1− throughout (Fig. 2C; supplementary material Fig. S1A,B). These data indicate that NGN3+ cells transit to KIT+ cells rapidly and efficiently in response to RA.Fig. 2.

Bottom Line: Ectopic expression of RARγ was sufficient to induce GFRα1(+) cells to directly differentiate to KIT(+) cells without transiting the NGN3(+) state.Therefore, RARγ plays key roles in the differentiation competence of NGN3(+) cells.We propose a novel mechanism of stem cell fate selection in an open niche environment whereby undifferentiated cells show heterogeneous competence to differentiate in response to ubiquitously distributed differentiation-inducing signals.

View Article: PubMed Central - PubMed

Affiliation: Division of Germ Cell Biology, National Institute for Basic Biology, National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji, Okazaki 444-8787, Japan Department of Basic Biology, School of Life Science, Graduate University for Advanced Studies (Sokendai), 5-1 Higashiyama, Myodaiji, Okazaki 444-8787, Japan.

Show MeSH
Related in: MedlinePlus