scholarly journals Lysosome Dynamic Properties during Neuronal Stem Cell Differentiation Studied by Spatiotemporal Fluctuation Spectroscopy and Organelle Tracking

2020 ◽  
Vol 21 (9) ◽  
pp. 3397 ◽  
Author(s):  
William Durso ◽  
Manuella Martins ◽  
Laura Marchetti ◽  
Federico Cremisi ◽  
Stefano Luin ◽  
...  
Keyword(s):  
Stem Cell ◽  
Dynamic Properties ◽  
Mean Square Displacement ◽  
Stem Cell Differentiation ◽  
Average Diameter ◽  
Single Particle Tracking ◽  
Dynamic Responses ◽  
Embryonic Cells ◽  
Dynamic Features ◽  
Neuronal Stem Cell

We investigated lysosome dynamics during neuronal stem cell (NSC) differentiation by two quantitative and complementary biophysical methods based on fluorescence: imaging-derived mean square displacement (iMSD) and single-particle tracking (SPT). The former extracts the average dynamics and size of the whole population of moving lysosomes directly from imaging, with no need to calculate single trajectories; the latter resolves the finest heterogeneities and dynamic features at the single-lysosome level, which are lost in the iMSD analysis. In brief, iMSD analysis reveals that, from a structural point of view, lysosomes decrement in size during NSC differentiation, from 1 μm average diameter in the embryonic cells to approximately 500 nm diameter in the fully differentiated cells. Concomitantly, iMSD analysis highlights modification of key dynamic parameters, such as the average local organelle diffusivity and anomalous coefficient, which may parallel cytoskeleton remodeling during the differentiation process. From average to local, SPT allows mapping heterogeneous dynamic responses of single lysosomes in different districts of the cells. For instance, a dramatic decrease of lysosomal transport in the soma is followed by a rapid increase of transport in the projections at specific time points during neuronal differentiation, an observation compatible with the hypothesis that lysosomal active mobilization shifts from the soma to the newborn projections. Our combined results provide new insight into the lysosome size and dynamics regulation throughout NSC differentiation, supporting new functions proposed for this organelle.

Dynamics and Traffic for Transfecting Exogenous MicroRNA as Studied by Live-Cell Microscopy

2021 ◽  
Vol 17 (8) ◽  
pp. 1647-1653
Author(s):  
Ke Yang ◽  
Yuanyuan Wang ◽  
Bo Sun ◽  
Tian Tian ◽  
Zhu Dai ◽  
...  
Keyword(s):  
Dynamic Properties ◽  
Mean Square Displacement ◽  
Small Region ◽  
Single Particle Tracking ◽  
Live Cell ◽  
Calculation Results ◽  
Long Time ◽  
Live Cell Microscopy ◽  
Cell Microscopy

MicroRNA (miRNA) has emerged as an important gene-regulator that shows great potential in gene therapy because of its unique roles in gene-regulation. However, the knowledge on their function and transportation in vivo is still lacking, and there are limited obvious evidences to define intracellular transportation of miRNA. In this study, the dynamics of exogenous miR-21 transfected into HeLa cells was traced by live-cell microscopy. Their transportation at key time points was recorded and dynamic properties were analyzed by single particle tracking (SPT) and mean square displacement (MSD) calculation. Results showed that the exogenous miRNAs bounded to cells quickly and went through lysosome into cytosol, where they were subsequently recruited into p-body. They finally were degraded, otherwise went back to cytosol in some way. Long time observation and analysis of motion mode showed that the miRNAs were confined in a small region and their motion modes were flexible in different intracellular microenvironment after entering the cells.

scholarly journals APLP2 regulates neuronal stem cell differentiation during cortical development

2013 ◽  
Vol 126 (5) ◽  
pp. 1268-1277 ◽  
Author(s):  
S. A. M. Shariati ◽  
P. Lau ◽  
B. A. Hassan ◽  
U. Muller ◽  
C. G. Dotti ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cell Differentiation ◽  
Cortical Development ◽  
Stem Cell Differentiation ◽  
Neuronal Stem Cell

In Vitro Effects of Chronic Spirolide Treatment on Human Neuronal Stem Cell Differentiation and Cholinergic System Development

2018 ◽  
Vol 9 (6) ◽  
pp. 1441-1452 ◽  
Author(s):  
Andrea Boente-Juncal ◽  
Aida G. Méndez ◽  
Carmen Vale ◽  
Mercedes R. Vieytes ◽  
Luis M. Botana
Keyword(s):  
Stem Cell ◽  
Cell Differentiation ◽  
Cholinergic System ◽  
System Development ◽  
Stem Cell Differentiation ◽  
Neuronal Stem Cell ◽  
In Vitro Effects

ERK-mediated production of neurotrophic factors by astrocytes promotes neuronal stem cell differentiation by erythropoietin

2006 ◽  
Vol 339 (4) ◽  
pp. 1021-1028 ◽  
Author(s):  
Mi Hee Park ◽  
Sang Min Lee ◽  
Jae Woong Lee ◽  
Dong Ju Son ◽  
Dong Cheul Moon ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cell Differentiation ◽  
Neurotrophic Factors ◽  
Stem Cell Differentiation ◽  
Neuronal Stem Cell

scholarly journals Decoding the mechanisms underlying cell-fate decision-making during stem cell differentiation by random circuit perturbation

2020 ◽  
Vol 17 (169) ◽  
pp. 20200500
Author(s):  
Bin Huang ◽  
Mingyang Lu ◽  
Madeline Galbraith ◽  
Herbert Levine ◽  
Jose N. Onuchic ◽  
...  
Keyword(s):  
Decision Making ◽  
Stem Cells ◽  
Stem Cell ◽  
Cell Fate ◽  
Expression Patterns ◽  
Stem Cell Differentiation ◽  
Computational Method ◽  
Embryonic Cells ◽  
Fate Decision ◽  
Stem Cell Populations

Stem cells can precisely and robustly undergo cellular differentiation and lineage commitment, referred to as stemness. However, how the gene network underlying stemness regulation reliably specifies cell fates is not well understood. To address this question, we applied a recently developed computational method, ra ndom ci rcuit pe rturbation (RACIPE), to a nine-component gene regulatory network (GRN) governing stemness, from which we identified robust gene states. Among them, four out of the five most probable gene states exhibit gene expression patterns observed in single mouse embryonic cells at 32-cell and 64-cell stages. These gene states can be robustly predicted by the stemness GRN but not by randomized versions of the stemness GRN. Strikingly, we found a hierarchical structure of the GRN with the Oct4/Cdx2 motif functioning as the first decision-making module followed by Gata6/Nanog. We propose that stem cell populations, instead of being viewed as all having a specific cellular state, can be regarded as a heterogeneous mixture including cells in various states. Upon perturbations by external signals, stem cells lose the capacity to access certain cellular states, thereby becoming differentiated. The new gene states and key parameters regulating transitions among gene states proposed by RACIPE can be used to guide experimental strategies to better understand differentiation and design reprogramming. The findings demonstrate that the functions of the stemness GRN is mainly determined by its well-evolved network topology rather than by detailed kinetic parameters.

scholarly journals APLP2 regulates neuronal stem cell differentiation during cortical development

Development ◽  
2013 ◽  
Vol 140 (11) ◽  
pp. e1-e1
Author(s):  
S. A. M. Shariati ◽  
P. Lau ◽  
B. A. Hassan ◽  
U. Muller ◽  
C. G. Dotti ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cell Differentiation ◽  
Cortical Development ◽  
Stem Cell Differentiation ◽  
Neuronal Stem Cell

Isolation of novel developmental genes from human germ cell, oocyte and embryo cDNA by differential display

2001 ◽  
Vol 13 (1) ◽  
pp. 51 ◽  
Author(s):  
Marilyn Monk ◽  
Cathy Holding ◽  
Tetsuya Goto
Keyword(s):  
Stem Cell ◽  
Early Development ◽  
Germ Cells ◽  
Differential Display ◽  
Primordial Germ Cells ◽  
Stem Cell Differentiation ◽  
Human Reproduction ◽  
Human Embryos ◽  
Embryonic Cells ◽  
Developmental Genes

Due to the difficulties inherent in research on human embryos, almost nothing is known about genes active in human early development. Although the human genome project will provide resources that theoretically provide access to every human gene, those genes specific to human early development may be difficult to define. Also, by definition, genes specific to early development will not be represented in cDNA databases derived from human somatic cells. Yet these unknown human developmental genes are likely to be of key importance for several areas of human health, including assisted reproduction and contraception, embryo stem cell research and tissue trans-plantation, ageing and cancer. In order to identify and isolate these human developmental genes, we have prepared amplified cDNA from human primordial germ cells, oocytes and embryos, and used differential display to compare patterns of gene expression in these embryonic cells and in the cells of somatic tissues of a 10-week human fetus. This paper reviews the highly sensitive procedures used to create amplified cDNA representing expressed genes in a single cell and the use of differential display to identify developmental genes. Several such genes have been isolated, but their full-length sequences and function are yet to be elucidated. Genes active in human early development are expected to play key roles in the maintenance of the archetypal stem cell state, potential immortality and the invasiveness of trophectoderm and primordial germ cells. They represent candidate genes regulating these functions for targeting in clinical research in human reproduction, stem cell differentiation and cancer.

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