TY - JOUR

T1 - Ouabain-induced gene expression changes in human ipsc-derived neuron culture expressing dopamine and camp-regulated phosphoprotein 32 and gaba receptors

AU - Lopachev, Alexander V.

AU - Lagarkova, Maria A.

AU - Lebedeva, Olga S.

AU - Ezhova, Margarita A.

AU - Kazanskaya, Rogneda B.

AU - Timoshina, Yulia A.

AU - Khutorova, Anastasiya V.

AU - Akkuratov, Evgeny E.

AU - Fedorova, Tatiana N.

AU - Gainetdinov, Raul R.

N1 - Funding Information: Funding: This study was supported by Russian Foundation for Basic Research (Grants N18-04-01321) and project ID:51143531 of the St. Petersburg State University, St. Petersburg, Russia. Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

PY - 2021/2/7

Y1 - 2021/2/7

N2 - Cardiotonic steroids (CTS) are specific inhibitors and endogenous ligands of a key enzyme in the CNS—the Na+, K+-ATPase, which maintains and creates an ion gradient on the plasma membrane of neurons. CTS cause the activation of various signaling cascades and changes in gene expression in neurons and other cell types. It is known that intracerebroventricular injection of cardiotonic steroid ouabain causes mania-like behavior in rodents, in part due to activation of dopamine-related signaling cascades in the dopamine and cAMP-regulated phosphoprotein 32 (DARPP-32) expressing medium spiny neurons in the striatum. Dopaminergic projections in the striatum innervate these GABAergic medium spiny neurons. The objective of this study was to assess changes in the expression of all genes in human iPSC-derived expressing DARPP-32 and GABA receptors neurons under the influence of ouabain. We noted a large number of statistically significant upregulated and downregulated genes after a 16-h incubation with non-toxic concentration (30 nM) of ouabain. These changes in the transcriptional activity were accomplished with activation of MAP-kinase ERK1/2 and transcriptional factor cAMP response element-binding protein (CREB). Thus, it can be concluded that 30 nM ouabain incubated for 16 h with human iPSC-derived expressing DARPP-32 and GABA receptors neurons activates genes associated with neuronal maturation and synapse formation, by increasing the expression of genes associated with translation, vesicular transport, and increased electron transport chain function. At the same time, the expression of genes associated with proliferation, migration, and early development of neurons decreases. These data indicate that non-toxic concentrations of ouabain may induce neuronal maturation, neurite growth, and increased synaptogenesis in dopamine-receptive GABAergic neurons, suggesting formation of plasticity and the establishment of new neuronal junctions.

AB - Cardiotonic steroids (CTS) are specific inhibitors and endogenous ligands of a key enzyme in the CNS—the Na+, K+-ATPase, which maintains and creates an ion gradient on the plasma membrane of neurons. CTS cause the activation of various signaling cascades and changes in gene expression in neurons and other cell types. It is known that intracerebroventricular injection of cardiotonic steroid ouabain causes mania-like behavior in rodents, in part due to activation of dopamine-related signaling cascades in the dopamine and cAMP-regulated phosphoprotein 32 (DARPP-32) expressing medium spiny neurons in the striatum. Dopaminergic projections in the striatum innervate these GABAergic medium spiny neurons. The objective of this study was to assess changes in the expression of all genes in human iPSC-derived expressing DARPP-32 and GABA receptors neurons under the influence of ouabain. We noted a large number of statistically significant upregulated and downregulated genes after a 16-h incubation with non-toxic concentration (30 nM) of ouabain. These changes in the transcriptional activity were accomplished with activation of MAP-kinase ERK1/2 and transcriptional factor cAMP response element-binding protein (CREB). Thus, it can be concluded that 30 nM ouabain incubated for 16 h with human iPSC-derived expressing DARPP-32 and GABA receptors neurons activates genes associated with neuronal maturation and synapse formation, by increasing the expression of genes associated with translation, vesicular transport, and increased electron transport chain function. At the same time, the expression of genes associated with proliferation, migration, and early development of neurons decreases. These data indicate that non-toxic concentrations of ouabain may induce neuronal maturation, neurite growth, and increased synaptogenesis in dopamine-receptive GABAergic neurons, suggesting formation of plasticity and the establishment of new neuronal junctions.

KW - Cardiotonic steroids

KW - Dopamine

KW - GABA

KW - Gene expression

KW - IPSC

KW - Na,K-ATPase

KW - Neurons

KW - Ouabain

KW - RNA-seq

KW - Transcriptome

KW - Na+,K+-ATPase

UR - http://www.scopus.com/inward/record.url?scp=85100984961&partnerID=8YFLogxK

UR - https://www.mendeley.com/catalogue/91c3c851-88cf-36ef-8cff-d64182a02c40/

U2 - 10.3390/brainsci11020203

DO - 10.3390/brainsci11020203

M3 - Article

AN - SCOPUS:85100984961

VL - 11

JO - Brain Sciences

JF - Brain Sciences

SN - 2076-3425

IS - 2

M1 - 203

ER -