Early Developmental PMCA2b Expression Protects From Ketamine-Induced Apoptosis and GABA Impairments in Differentiating Hippocampal Progenitor Cells

Malwina Lisek; Joanna Mackiewicz; Marta Sobolczyk; Bozena Ferenc; Feng Guo; Ludmila Zylinska; Tomasz Boczek

doi:10.3389/fncel.2022.890827

Early Developmental PMCA2b Expression Protects From Ketamine-Induced Apoptosis and GABA Impairments in Differentiating Hippocampal Progenitor Cells

Front Cell Neurosci. 2022 May 23:16:890827. doi: 10.3389/fncel.2022.890827. eCollection 2022.

Authors

Malwina Lisek¹, Joanna Mackiewicz¹, Marta Sobolczyk¹, Bozena Ferenc¹, Feng Guo², Ludmila Zylinska¹, Tomasz Boczek¹

Affiliations

¹ Department of Molecular Neurochemistry, Medical University of Lodz, Łódz, Poland.
² Department of Pharmaceutical Toxicology, China Medical University, Shenyang, China.

Abstract

PMCA2 is not expressed until the late embryonic state when the control of subtle Ca²⁺ fluxes becomes important for neuronal specialization. During this period, immature neurons are especially vulnerable to degenerative insults induced by the N-methyl-D-aspartate (NMDA) receptor blocker, ketamine. As H19-7 hippocampal progenitor cells isolated from E17 do not express the PMCA2 isoform, they constitute a valuable model for studying its role in neuronal development. In this study, we demonstrated that heterologous expression of PMCA2b enhanced the differentiation of H19-7 cells and protected from ketamine-induced death. PMCA2b did not affect resting [Ca²⁺]_c in the presence or absence of ketamine and had no effect on the rate of Ca²⁺ clearance following membrane depolarization in the presence of the drug. The upregulation of endogenous PMCA1 demonstrated in response to PMCA2b expression as well as ketamine-induced PMCA4 depletion were indifferent to the rate of Ca²⁺ clearance in the presence of ketamine. Yet, co-expression of PMCA4b and PMCA2b was able to partially restore Ca²⁺ extrusion diminished by ketamine. The profiling of NMDA receptor expression showed upregulation of the NMDAR1 subunit in PMCA2b-expressing cells and increased co-immunoprecipitation of both proteins following ketamine treatment. Further microarray screening demonstrated a significant influence of PMCA2b on GABA signaling in differentiating progenitor cells, manifested by the unique regulation of several genes key to the GABAergic transmission. The overall activity of glutamate decarboxylase remained unchanged, but Ca²⁺-induced GABA release was inhibited in the presence of ketamine. Interestingly, PMCA2b expression was able to reverse this effect. The mechanism of GABA secretion normalization in the presence of ketamine may involve PMCA2b-mediated inhibition of GABA transaminase, thus shifting GABA utilization from energetic purposes to neurosecretion. In this study, we show for the first time that developmentally controlled PMCA expression may dictate the pattern of differentiation of hippocampal progenitor cells. Moreover, the appearance of PMCA2 early in development has long-standing consequences for GABA metabolism with yet an unpredictable influence on GABAergic neurotransmission during later stages of brain maturation. In contrast, the presence of PMCA2b seems to be protective for differentiating progenitor cells from ketamine-induced apoptotic death.

Keywords: GABA metabolism; calcium; hippocampal progenitor cells; ketamine; neuronal differentiation; plasma membrane Ca2+-ATPase (PMCA).