Calcium signaling mediates five types of cell morphological changes to form neural rosettes

Hříbková, Hana; Grabiec, Marta; Klemová, Dobromila; Slaninová, Iva; Sun, Yin-Biao

doi:10.1242/jcs.206896

Cited by 50 publications

(28 citation statements)

References 68 publications

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“…Additional immunolocalization analyses reveal that recycling (RAB11) and early (RAB5) endosomes are localized adjacent to the apical cortex, and Golgi (GM130) and centrosomes (γ-TUBULIN) are localized at the apical pole of the cells ( Figure 2C ), further confirming the apical nature of the central domain. Interestingly, 3D reconstruction of NPC rosettes stained for the apical marker PODXL reveals that cells completely surround the lumen ( Figure 2D ), confirming recent studies ( Hribkova et al, 2018 ). Indeed, the apical region of each cell is studded with a single primary cilium (ARL13B, Figure 2E ); this is in accord with the localization of the centrosomes (γ-TUBULIN, Figure 2C ).…”

Section: Resultssupporting

confidence: 88%

Effect of Cell Spreading on Rosette Formation by Human Pluripotent Stem Cell-Derived Neural Progenitor Cells

Townshend

Shao

Wang

et al. 2020

Front. Cell Dev. Biol.

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Neural rosettes (NPC rosettes) are radially arranged groups of cells surrounding a central lumen that arise stochastically in monolayer cultures of human pluripotent stem cell (hPSC)-derived neural progenitor cells (NPC). Since NPC rosette formation is thought to mimic cell behavior in the early neural tube, these rosettes represent important in vitro models for the study of neural tube morphogenesis. However, using current protocols, NPC rosette formation is not synchronized and results are inconsistent among different hPSC lines, hindering quantitative mechanistic analyses and challenging live cell imaging. Here, we report a rapid and robust protocol to induce rosette formation within 6 h after evenly-sized “colonies” of NPC are generated through physical cutting of uniformly polarized NESTIN + /PAX6 + /PAX3 + /DACH1 + NPC monolayers. These NPC rosettes show apically polarized lumens studded with primary cilia. Using this assay, we demonstrate reduced lumenal size in the absence of PODXL , an important apical determinant recently identified as a candidate gene for juvenile Parkinsonism. Interestingly, time lapse imaging reveals that, in addition to radial organization and apical lumen formation, cells within cut NPC colonies initiate rapid basally-driven spreading. Further, using chemical, genetic and biomechanical tools, we show that NPC rosette morphogenesis requires this basal spreading activity and that spreading is tightly regulated by Rho/ROCK signaling. This robust and quantitative NPC rosette platform provides a sensitive system for the further investigation of cellular and molecular mechanisms underlying NPC rosette morphogenesis.

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Section: Resultssupporting

confidence: 88%

Effect of Cell Spreading on Rosette Formation by Human Pluripotent Stem Cell-Derived Neural Progenitor Cells

Townshend

Shao

Wang

et al. 2020

Front. Cell Dev. Biol.

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“…High resolution images show the formation of organized neural rosettes within the EBs at day 8 [1,11]. Alpha-tubulin (α-tubulin filaments) can be seen organized radially from the lumen (Figure 18C) [16][17][18][19]. These polarized neuroepithelium-like structures resemble neural tube formation and are the precursors to the formation of the brain lobules [10].…”

Section: Resultsmentioning

confidence: 99%

Spin∞ an improved miniaturized spinning bioreactor for the generation of human cerebral organoids from pluripotent stem cells

Romero-Morales

O’Grady

Balotin

et al. 2019

Preprint

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Three-dimensional (3D) brain organoids derived from human pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs), have become a powerful system to study early development events and to model human disease. Cerebral organoids are generally produced in static culture or in a culture vessel with active mixing, and the two most widely used systems for mixing are a large spinning flask and a miniaturized multi-well spinning bioreactor (also known as Spin Omega (SpinΩ)). The SpinΩ provides a system that is amenable to drug testing, has increased throughput and reproducibility, and utilizes less culture media. However, technical limitations of this system include poor stability of select components and an elevated risk of contamination due to the inability to sterilize the device preassembled. Here, we report a new design of the miniaturized bioreactor system, which we term Spin∞ that overcomes these concerns to permit long-term experiments.Brain organoids are three-dimensional (3D) structures formed from neural stem cells (NSCs) derived from human pluripotent stem cells (hPSCs) that can effectively model human brain development up to 12-14 weeks post-conception [1-4], a time period which includes critical patterning events in the cerebral cortex and other brain regions [5,6]. On a cellular level, brain organoids show a high level of similarity to the in vivo developing human brain in the early stages of development, including progenitor zones (ventricular zone and subventricular zone consisting of PAX6+/SOX2+ NSCs) that form around central lumens [1,4].These 3D organoid cultures therefore provide a robust system amenable to extended cultivation and manipulation, which makes them a useful tool to model development and disease in the context of the complex brain microenvironment [7][8][9].Recently, two protocols have been published on enhancing cortical plate formation within hPSC-derived cerebral organoids: one using large spinner flasks and microfilaments as a solid support [1,10] and another that uses a miniature spinning bioreactor termed Spin Omega (SpinΩ) [11,12], which consists of 3D printed gears and paddles driven by a single electric motor. The SpinΩ provide an accessible and versatile format for culturing brain-region-specific organoids due to its reduced incubator footprint, decreased media consumption, and increased throughput, but several technical caveats limit its use in long-term experiments, most prominently the choice of components used to fabricate the device and the design of the device with respect to limiting the chances of contamination and mechanical failure. Here, we redesigned the SpinΩ to overcome these problems, leading to the creation of a device that we have termed Spinfinity (Spin∞).

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“…Tight junctions are essential for establishing the selectively permeable barrier between neighboring cells, tight junctions formed between brain capillary endothelial cells help creating blood-brain barrier that act to protect the brain from harmful substances and maintain brain homeostasis, modi ed tight junction protein expression and disrupted blood-brain barrier structure are observed in cerebral pathologies [22,23]. Calcium signaling regulates nearly every aspect of neural development, including neural induction, neural rosettes formation, neural progenitor cells proliferation, neuronal migration and differentiation [24][25][26], speci c synaptic connections development, and impairments in calcium-dependent network maturation are associated with neurodevelopment disorders [27].…”

Section: Discussionmentioning

confidence: 99%

Maternal Folic Acid Impacts DNA Methylation Profile in Male Rat Offspring Associated With Neurodevelopment and Learning/Memory Abilities

Wang

Zhu

et al. 2020

Preprint

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Background: Periconceptional folic acid (FA) supplementation not only reduces the incidence of neural tube defects, but also improves cognitive performances in offspring. However, the genes or pathways that are epigenetically regulated by FA in neurodevelopment were rarely reported. Methods: To elucidate the underlying mechanism, the effect of FA on the methylation profiles in brain tissue of male rat offspring was assessed by methylated DNA immunoprecipitation chip. Differentially methylated genes (DMGs) and gene network analysis were identified using DAVID and KEGG pathway analysis. Results: Compared with the FA-N group, 1939 DMGs were identified in the FA-D group, and 1498 DMGs were identified in the FA-S group, among which 298 DMGs were overlapped. The pathways associated with neurodevelopment and learning/memory abilities were differentially methylated in response to maternal FA intake during pregnancy, and there were some identical and distinctive potential mechanisms under FA deficiency or supplemented conditions. Conclusions: In conclusion, genes and pathways associated with neurodevelopment and learning/memory abilities were differentially methylated in male rat offspring in response to maternal FA deficiency or supplementation during pregnancy.

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Calcium signaling mediates five types of cell morphological changes to form neural rosettes

Cited by 50 publications

References 68 publications

Effect of Cell Spreading on Rosette Formation by Human Pluripotent Stem Cell-Derived Neural Progenitor Cells

Effect of Cell Spreading on Rosette Formation by Human Pluripotent Stem Cell-Derived Neural Progenitor Cells

Spin∞ an improved miniaturized spinning bioreactor for the generation of human cerebral organoids from pluripotent stem cells

Maternal Folic Acid Impacts DNA Methylation Profile in Male Rat Offspring Associated With Neurodevelopment and Learning/Memory Abilities

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