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Insulin biosynthesis in neuronal progenitors derived from adult hippocampus and the olfactory bulb.

Kuwabara T, Kagalwala MN, Onuma Y, Ito Y, Warashina M, Terashima K, Sanosaka T, Nakashima K, Gage FH, Asashima M - EMBO Mol Med (2011)

Bottom Line: Paracrine Wnt3 plays an essential role in promoting the active expression of insulin in both hippocampal and OB-derived neural stem cells.We also show that adult neural progenitors derived from DB animals retain the ability to give rise to insulin-producing cells and that grafting neuronal progenitors into the pancreas of DB animals reduces glucose levels.This study provides an example of a simple and direct use of adult stem cells from one organ to another, without introducing additional inductive genes.

View Article: PubMed Central - PubMed

Affiliation: Research Center for Stem Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Science City, Japan. t.warashina@aist.go.jp

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Adult NSCs derived from HPC and OB give rise to neurons expressing insulinThe adult NSCs isolated from HPC and OB. Culture image of the proliferating hippocampal NSCs (HPC NSC, left) and NSCs derived from OB (OB NSC, right) of 7- to 8-week-old female Fisher 344 rats.In vitro differentiated adult HPC- and OB neurons. When the culture was exposed to neuron differentiation conditions (RA + FSK + KCl), both HPC and OB cells extended neuritis (HPC neuron, left; OB neuron, right).Gene expression profiles of HPC NSC, OB NSC, HPC neuron and OB neuron that were measured using Agilent rat genome microarrays. Typical genes for neuronal cell lineages, neural crest marker, pancreatic genes and mature neuron marker were analysed and the heat map using average-linkage hierarchical clustering is shown.Induction of expression levels of insulin in HPC- and OB NSCs differentiating into neurons. The expression levels of proinsulin 1 and NeuroD1 were determined by Q-PCR analysis. Each mRNA value was normalized to GAPDH and then plotted.Down-regulation of the neuronal expression of proinsulin 1 mRNAs with NeuroD1 siRNA. The neuronal induction of proinsulin 1 mRNA using siRNAs for NeuroD1, MafA, Pdx1 or Neurogenin 3 in HPC and OB neurons was examined in qRT-PCR analysis. Contol siRNA (scrambled siRNA) was transfected similarly. For quantification, the level of the control siRNA in NSCs group was set at 1, and the experimental siRNAs in neurons were then normalized accordingly.
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fig03: Adult NSCs derived from HPC and OB give rise to neurons expressing insulinThe adult NSCs isolated from HPC and OB. Culture image of the proliferating hippocampal NSCs (HPC NSC, left) and NSCs derived from OB (OB NSC, right) of 7- to 8-week-old female Fisher 344 rats.In vitro differentiated adult HPC- and OB neurons. When the culture was exposed to neuron differentiation conditions (RA + FSK + KCl), both HPC and OB cells extended neuritis (HPC neuron, left; OB neuron, right).Gene expression profiles of HPC NSC, OB NSC, HPC neuron and OB neuron that were measured using Agilent rat genome microarrays. Typical genes for neuronal cell lineages, neural crest marker, pancreatic genes and mature neuron marker were analysed and the heat map using average-linkage hierarchical clustering is shown.Induction of expression levels of insulin in HPC- and OB NSCs differentiating into neurons. The expression levels of proinsulin 1 and NeuroD1 were determined by Q-PCR analysis. Each mRNA value was normalized to GAPDH and then plotted.Down-regulation of the neuronal expression of proinsulin 1 mRNAs with NeuroD1 siRNA. The neuronal induction of proinsulin 1 mRNA using siRNAs for NeuroD1, MafA, Pdx1 or Neurogenin 3 in HPC and OB neurons was examined in qRT-PCR analysis. Contol siRNA (scrambled siRNA) was transfected similarly. For quantification, the level of the control siRNA in NSCs group was set at 1, and the experimental siRNAs in neurons were then normalized accordingly.

Mentions: HPC NSCs were round and retained this shape when expanded as a monolayer (Fig 3A, left). OB NSCs grew as heterogeneous forms with adherent property and neurosphere morphologies (Fig 3A, right). Under neuron differentiation conditions, the cell morphologies changed markedly: both HPC and OB NSCs extended prolonged neurites (Fig 3B). Gene expression profiles were measured using Agilent rat genome microarrays (Table 1 of Supporting information). HPC NSC, OB NSC and HPC and OB neuron expression profiles were grouped by hierarchical clustering, and correlation coefficients were computed for all pair-wise comparisons (Fig S5 of Supporting information). Both HPC NSCs and OB NSCs expressed the NSC marker Sox2, the radial glial cell marker nestin and the neural progenitor markers Olig2 and Sox1 (NSC lineage; Fig 3C). NSC marker gene expression was down-regulated in HPC and OB neurons (HPC Neu and OB Neu in Fig 3C), and neuronal marker gene expression was up-regulated (mature neuron marker; Fig 3C), indicating their successful commitment into neuronal lineages.


Insulin biosynthesis in neuronal progenitors derived from adult hippocampus and the olfactory bulb.

Kuwabara T, Kagalwala MN, Onuma Y, Ito Y, Warashina M, Terashima K, Sanosaka T, Nakashima K, Gage FH, Asashima M - EMBO Mol Med (2011)

Adult NSCs derived from HPC and OB give rise to neurons expressing insulinThe adult NSCs isolated from HPC and OB. Culture image of the proliferating hippocampal NSCs (HPC NSC, left) and NSCs derived from OB (OB NSC, right) of 7- to 8-week-old female Fisher 344 rats.In vitro differentiated adult HPC- and OB neurons. When the culture was exposed to neuron differentiation conditions (RA + FSK + KCl), both HPC and OB cells extended neuritis (HPC neuron, left; OB neuron, right).Gene expression profiles of HPC NSC, OB NSC, HPC neuron and OB neuron that were measured using Agilent rat genome microarrays. Typical genes for neuronal cell lineages, neural crest marker, pancreatic genes and mature neuron marker were analysed and the heat map using average-linkage hierarchical clustering is shown.Induction of expression levels of insulin in HPC- and OB NSCs differentiating into neurons. The expression levels of proinsulin 1 and NeuroD1 were determined by Q-PCR analysis. Each mRNA value was normalized to GAPDH and then plotted.Down-regulation of the neuronal expression of proinsulin 1 mRNAs with NeuroD1 siRNA. The neuronal induction of proinsulin 1 mRNA using siRNAs for NeuroD1, MafA, Pdx1 or Neurogenin 3 in HPC and OB neurons was examined in qRT-PCR analysis. Contol siRNA (scrambled siRNA) was transfected similarly. For quantification, the level of the control siRNA in NSCs group was set at 1, and the experimental siRNAs in neurons were then normalized accordingly.
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fig03: Adult NSCs derived from HPC and OB give rise to neurons expressing insulinThe adult NSCs isolated from HPC and OB. Culture image of the proliferating hippocampal NSCs (HPC NSC, left) and NSCs derived from OB (OB NSC, right) of 7- to 8-week-old female Fisher 344 rats.In vitro differentiated adult HPC- and OB neurons. When the culture was exposed to neuron differentiation conditions (RA + FSK + KCl), both HPC and OB cells extended neuritis (HPC neuron, left; OB neuron, right).Gene expression profiles of HPC NSC, OB NSC, HPC neuron and OB neuron that were measured using Agilent rat genome microarrays. Typical genes for neuronal cell lineages, neural crest marker, pancreatic genes and mature neuron marker were analysed and the heat map using average-linkage hierarchical clustering is shown.Induction of expression levels of insulin in HPC- and OB NSCs differentiating into neurons. The expression levels of proinsulin 1 and NeuroD1 were determined by Q-PCR analysis. Each mRNA value was normalized to GAPDH and then plotted.Down-regulation of the neuronal expression of proinsulin 1 mRNAs with NeuroD1 siRNA. The neuronal induction of proinsulin 1 mRNA using siRNAs for NeuroD1, MafA, Pdx1 or Neurogenin 3 in HPC and OB neurons was examined in qRT-PCR analysis. Contol siRNA (scrambled siRNA) was transfected similarly. For quantification, the level of the control siRNA in NSCs group was set at 1, and the experimental siRNAs in neurons were then normalized accordingly.
Mentions: HPC NSCs were round and retained this shape when expanded as a monolayer (Fig 3A, left). OB NSCs grew as heterogeneous forms with adherent property and neurosphere morphologies (Fig 3A, right). Under neuron differentiation conditions, the cell morphologies changed markedly: both HPC and OB NSCs extended prolonged neurites (Fig 3B). Gene expression profiles were measured using Agilent rat genome microarrays (Table 1 of Supporting information). HPC NSC, OB NSC and HPC and OB neuron expression profiles were grouped by hierarchical clustering, and correlation coefficients were computed for all pair-wise comparisons (Fig S5 of Supporting information). Both HPC NSCs and OB NSCs expressed the NSC marker Sox2, the radial glial cell marker nestin and the neural progenitor markers Olig2 and Sox1 (NSC lineage; Fig 3C). NSC marker gene expression was down-regulated in HPC and OB neurons (HPC Neu and OB Neu in Fig 3C), and neuronal marker gene expression was up-regulated (mature neuron marker; Fig 3C), indicating their successful commitment into neuronal lineages.

Bottom Line: Paracrine Wnt3 plays an essential role in promoting the active expression of insulin in both hippocampal and OB-derived neural stem cells.We also show that adult neural progenitors derived from DB animals retain the ability to give rise to insulin-producing cells and that grafting neuronal progenitors into the pancreas of DB animals reduces glucose levels.This study provides an example of a simple and direct use of adult stem cells from one organ to another, without introducing additional inductive genes.

View Article: PubMed Central - PubMed

Affiliation: Research Center for Stem Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Science City, Japan. t.warashina@aist.go.jp

Show MeSH
Related in: MedlinePlus