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Expression and Functional Properties of NMDA and GABAA Receptors during Differentiation of Human Induced Pluripotent Stem Cells into Ventral Mesencephalic Neurons

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Abstract

Ionotropic glutamate and GABA receptors regulate the differentiation and determine the functional properties of mature neurons. Both insufficient and excessive activity of these neurotransmission systems are associated with various nervous system diseases. Our knowledge regarding the expression profiles of these receptors and the mechanisms of their regulation during the differentiation of specialized human neuron subtypes is limited. Here the expression profiles of the NMDA and GABAA receptor subunits were explored during in vitro differentiation of human induced pluripotent stem cells (iPSCs) into ventral mesencephalic neurons. The correlation between the neuronal maturation and the expression dynamics of these genes was investigated, and the functional activity of these receptors was assessed by calcium imaging. The role of NMDA and GABAA receptors in neurite outgrowth and the development of spontaneous activity was analyzed using the viral transduction of neural progenitors with the reporter genes TagGFP and TagRFP. The data indicate that agonists of the investigated receptors can be employed for optimization of existing protocols for neural differentiation of iPSCs, in particular for acceleration of neuronal maturation.

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Abbreviations

BDNF:

brain derived nerve growth factor

(c)NPs:

(committed) neural progenitors

eNPs:

early neural progenitors

ESCs:

embryonic stem cells

GABA:

α-aminobutyric acid

GABAA-R:

GABAA receptors

GDNF:

glial cell derived nerve growth factor

iPSCs:

induced pluripotent stem cells

NMDA:

N-methyl-D-aspartic acid

NMDA-R:

NMDA receptors

RT-PCR:

reverse transcription polymerase chain reaction

TD:

terminal differentiation

VM neurons:

ventral mesencephalic neurons

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Authors and Affiliations

  1. Institute of Molecular Genetics, Russian Academy of Sciences, 123182, Moscow, Russia

    S. A. Antonov, E. V. Novosadova, A. G. Kobylyansky, V. Z. Tarantul & I. A. Grivennikov

  2. Research Center of Neurology, 125367, Moscow, Russia

    S. N. Illarioshkin

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  1. S. A. Antonov
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  2. E. V. Novosadova
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  3. A. G. Kobylyansky
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  4. S. N. Illarioshkin
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  5. V. Z. Tarantul
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  6. I. A. Grivennikov
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Correspondence to S. A. Antonov.

Additional information

Published in Russian in Biokhimiya, 2019, Vol. 84, No. 3, pp. 436–447.

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Antonov, S.A., Novosadova, E.V., Kobylyansky, A.G. et al. Expression and Functional Properties of NMDA and GABAA Receptors during Differentiation of Human Induced Pluripotent Stem Cells into Ventral Mesencephalic Neurons. Biochemistry Moscow 84, 310–320 (2019). https://doi.org/10.1134/S0006297919030131

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