Central and Peripheral Nervous System Progenitors Derived from Human Pluripotent Stem Cells Reveal a Unique Temporal and Cell-Type Specific Expression of PMCAs

Abstract

The P-type ATPases family consists of ion and lipid transporters. Their unique diversity in function and expression is critical for normal development. In this study we investigated human pluripotent stem cells (hPSC) and different neural progenitor states to characterize the expression of the plasma membrane calcium ATPases (PMCAs) during human neural development and in mature mesencephalic dopaminergic (mesDA) neurons. Our RNA sequencing data identified a dynamic change in ATPase expression correlating with the differentiation time of the neural progenitors, which was independent of the neuronal progenitor type. Expression of ATP2B1 and ATP2B4 were the most abundantly expressed, in accordance with their main role in Ca2+ regulation and we observed all of the PMCAs to have a subcellular punctate localization. Interestingly in hPSCs ATP2B1 and ATP2B3 were highly expressed in a cell cycle specific manner and ATP2B2 and ATP2B4 were highly expressed in a hPSC sub-population. In neural rosettes a strong apical PMCA expression was identified in the luminal region. Lastly, we confirmed all PMCAs to be expressed in mesDA neurons, however at varying levels. Our results reveal that PMCA expression dynamically changes during stem cell differentiation and highlights the diverging needs of cell populations to regulate and properly integrate Ca2+ changes, which can ultimately correspond to changes in specific stem cell transcription states.

A Circular RNA Expressed from the FAT3 Locus Regulates Neural Development

“…To assess potential roles of circRNAs in neural development, we first characterized the expression landscape of circRNAs in a 2D model during early neurogenesis. H9 human embryonic stem cells (hESCs) were differentiated into neuroepithelial or -mesodermal progenitor cells (NEPs/NMPS) and further into rostral and caudal neural progenitor cells (rNPCs and cNPCs, respectively) for 11 days, as previously described [ 35 ] (Fig. 1 A).…”

“…Differentiations were performed in biological triplicates. B Heatmap showing expression levels of characteristic markers in hESCs, NEPs, rNPCs, NMPs, and cNPCs [ 35 ] that were log2-transformed (normalized counts + pseudocount) and converted to a z -scale; n = 3. C Column scatter plot of 417 high-abundance circRNAs detected in hESCs, NEPs, rNPCs, NMPs, and cNPCs; n = 3.…”

“…E Profiling of days 0 and 11 differentiated H9 hESCs using characteristic markers established in Fig. 1 B and as previously described [ 35 ]. Normalized DESeq2 counts were log2-transformed (normalized counts + pseudo counts) and converted to a z-scale.…”

“…H9 hESCs were transferred to vitronectin-coated 4-well plates in N2B27 medium (1:1 mix of neurobasal medium and DMEM/F12 medium, 1% insulin/transferrin/selenium, 1 × N2 supplement, 1 × retinol-free B27 supplement, 0.3% glucose, and 1% p/s (Thermo Fisher Scientific)) for 11 days of differentiation. As previously described [ 35 ], for rostral neural progenitor cell (NPC) differentiation, SB431542 (SB, 10 µM; Tocris, Bristol, UK) and LDN193189 (LDN, 0.1 µM; STEMCELL Technologies) were added to the media for 4 days. The cells were further cultured with SB, LDN, and FGF2 (20 ng/ml; STEMCELL Technologies) until day seven.…”

Biallelic ATP2B1 variants as a likely cause of a novel neurodevelopmental malformation syndrome with primary hypoparathyroidism

“…Murine studies establish marked surges in Atp2b1 expression during embryogenesis at the early stage of differentiation into neural progenitor cells, during mitosis and later stages of neural tube formation, postulated to promote neural tube closure in neuronal migration [ 7 , 12 ]. It is therefore plausible that significant disruption of ATP2B1 function in human brain predisposes to a neurodevelopmental and malformation phenotype.…”