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Generation of diverse neuronal subtypes in cloned populations of stem-like cells.

Varga BV, Hádinger N, Gócza E, Dulberg V, Demeter K, Madarász E, Herberth B - BMC Dev. Biol. (2008)

Bottom Line: Non-induced neural stem cells and self-renewing cells persisting in differentiated cultures, expressed "stemness genes" along with early embryonic anterior-dorsal positional genes, but did not express the investigated CNS region-specific genes.Mechanisms inherent to one-cell derived neural stem cell populations were sufficient to establish glutamatergic and GABAergic neuronal phenotypes but failed to manifest cathecolaminergic neurons.Interactions among progenies of one cell derived neural stem cells are sufficient for the activation of diverse region specific genes and initiate different routes of neuronal specification.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory of Cellular and Developmental Neurobiology, Institute of Experimental Medicine of Hungarian Academy of Sciences, Budapest, Hungary. vargab@koki.hu

ABSTRACT

Background: The central nervous tissue contains diverse subtypes of neurons with characteristic morphological and physiological features and different neurotransmitter phenotypes. The generation of neurons with defined neurotransmitter phenotypes seems to be governed by factors differently expressed along the anterior-posterior and dorsal-ventral body axes. The mechanisms of the cell-type determination, however, are poorly understood. Selected neuronal phenotypes had been generated from embryonic stem (ES) cells, but similar results were not obtained on more restricted neural stem cells, presumably due to the lack of homogeneous neural stem cell populations as a starting material.

Results: In the presented work, the establishment of different neurotransmitter phenotypes was investigated in the course of in vitro induced neural differentiation of a one-cell derived neuroectodermal cell line, in conjunction with the activation of various region-specific genes. For comparison, similar studies were carried out on the R1 embryonic stem (ES) and P19 multipotent embryonic carcinoma (EC) cells. In response to a short treatment with all-trans retinoic acid, all cell lines gave rise to neurons and astrocytes. Non-induced neural stem cells and self-renewing cells persisting in differentiated cultures, expressed "stemness genes" along with early embryonic anterior-dorsal positional genes, but did not express the investigated CNS region-specific genes. In differentiating stem-like cell populations, on the other hand, different region-specific genes, those expressed in non-overlapping regions along the body axes were activated. The potential for diverse regional specifications was induced in parallel with the initiation of neural tissue-type differentiation. In accordance with the wide regional specification potential, neurons with different neurotransmitter phenotypes developed. Mechanisms inherent to one-cell derived neural stem cell populations were sufficient to establish glutamatergic and GABAergic neuronal phenotypes but failed to manifest cathecolaminergic neurons.

Conclusion: The data indicate that genes involved in positional determination are activated along with pro-neuronal genes in conditions excluding any outside influences. Interactions among progenies of one cell derived neural stem cells are sufficient for the activation of diverse region specific genes and initiate different routes of neuronal specification.

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Neurons develop from ES, EC and NE-4C cells in response to treatment with retinoic acid. Cultures were stained for neuron-specific βIII-tubulin in a phase of differentiation corresponding to stage 4 of NE-4C development: the 15th day of induced differentiation of R1 (ES), the 4th day for P19 (EC) and the 7th day for NE-4C. Bars: 50 μm
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Figure 1: Neurons develop from ES, EC and NE-4C cells in response to treatment with retinoic acid. Cultures were stained for neuron-specific βIII-tubulin in a phase of differentiation corresponding to stage 4 of NE-4C development: the 15th day of induced differentiation of R1 (ES), the 4th day for P19 (EC) and the 7th day for NE-4C. Bars: 50 μm

Mentions: Morphological, cell biological and molecular changes were followed during retinoic acid (all-trans retinoic acid; RA) induced neural differentiation of R1(ES) [23], P19 (EC) [29,24,30]; and NE-4C embryonic neuroectodermal [22] stem cells. Large number of neurons was formed by all stem-like cell populations (Fig. 1) on a reproducible but cell line-dependent schedule. Bulk neuron formation was evident in P19 cultures by the 3rd day of RA-induction. For mass appearance of neurons, ES cells must have been first grown as aggregates and treated with RA for 4 days (EB4; see M&M), and then incubated in monolayer cultures for a further 7 days.


Generation of diverse neuronal subtypes in cloned populations of stem-like cells.

Varga BV, Hádinger N, Gócza E, Dulberg V, Demeter K, Madarász E, Herberth B - BMC Dev. Biol. (2008)

Neurons develop from ES, EC and NE-4C cells in response to treatment with retinoic acid. Cultures were stained for neuron-specific βIII-tubulin in a phase of differentiation corresponding to stage 4 of NE-4C development: the 15th day of induced differentiation of R1 (ES), the 4th day for P19 (EC) and the 7th day for NE-4C. Bars: 50 μm
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2556672&req=5

Figure 1: Neurons develop from ES, EC and NE-4C cells in response to treatment with retinoic acid. Cultures were stained for neuron-specific βIII-tubulin in a phase of differentiation corresponding to stage 4 of NE-4C development: the 15th day of induced differentiation of R1 (ES), the 4th day for P19 (EC) and the 7th day for NE-4C. Bars: 50 μm
Mentions: Morphological, cell biological and molecular changes were followed during retinoic acid (all-trans retinoic acid; RA) induced neural differentiation of R1(ES) [23], P19 (EC) [29,24,30]; and NE-4C embryonic neuroectodermal [22] stem cells. Large number of neurons was formed by all stem-like cell populations (Fig. 1) on a reproducible but cell line-dependent schedule. Bulk neuron formation was evident in P19 cultures by the 3rd day of RA-induction. For mass appearance of neurons, ES cells must have been first grown as aggregates and treated with RA for 4 days (EB4; see M&M), and then incubated in monolayer cultures for a further 7 days.

Bottom Line: Non-induced neural stem cells and self-renewing cells persisting in differentiated cultures, expressed "stemness genes" along with early embryonic anterior-dorsal positional genes, but did not express the investigated CNS region-specific genes.Mechanisms inherent to one-cell derived neural stem cell populations were sufficient to establish glutamatergic and GABAergic neuronal phenotypes but failed to manifest cathecolaminergic neurons.Interactions among progenies of one cell derived neural stem cells are sufficient for the activation of diverse region specific genes and initiate different routes of neuronal specification.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory of Cellular and Developmental Neurobiology, Institute of Experimental Medicine of Hungarian Academy of Sciences, Budapest, Hungary. vargab@koki.hu

ABSTRACT

Background: The central nervous tissue contains diverse subtypes of neurons with characteristic morphological and physiological features and different neurotransmitter phenotypes. The generation of neurons with defined neurotransmitter phenotypes seems to be governed by factors differently expressed along the anterior-posterior and dorsal-ventral body axes. The mechanisms of the cell-type determination, however, are poorly understood. Selected neuronal phenotypes had been generated from embryonic stem (ES) cells, but similar results were not obtained on more restricted neural stem cells, presumably due to the lack of homogeneous neural stem cell populations as a starting material.

Results: In the presented work, the establishment of different neurotransmitter phenotypes was investigated in the course of in vitro induced neural differentiation of a one-cell derived neuroectodermal cell line, in conjunction with the activation of various region-specific genes. For comparison, similar studies were carried out on the R1 embryonic stem (ES) and P19 multipotent embryonic carcinoma (EC) cells. In response to a short treatment with all-trans retinoic acid, all cell lines gave rise to neurons and astrocytes. Non-induced neural stem cells and self-renewing cells persisting in differentiated cultures, expressed "stemness genes" along with early embryonic anterior-dorsal positional genes, but did not express the investigated CNS region-specific genes. In differentiating stem-like cell populations, on the other hand, different region-specific genes, those expressed in non-overlapping regions along the body axes were activated. The potential for diverse regional specifications was induced in parallel with the initiation of neural tissue-type differentiation. In accordance with the wide regional specification potential, neurons with different neurotransmitter phenotypes developed. Mechanisms inherent to one-cell derived neural stem cell populations were sufficient to establish glutamatergic and GABAergic neuronal phenotypes but failed to manifest cathecolaminergic neurons.

Conclusion: The data indicate that genes involved in positional determination are activated along with pro-neuronal genes in conditions excluding any outside influences. Interactions among progenies of one cell derived neural stem cells are sufficient for the activation of diverse region specific genes and initiate different routes of neuronal specification.

Show MeSH
Related in: MedlinePlus