Spontaneous calcium influx and its roles in differentiation of spinal neurons in culture

Holliday, Janet; Spitzer, Nicholas C.

doi:10.1016/0012-1606(90)90098-4

Cited by 134 publications

(103 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although the involvement of Ca 2ϩ in neuronal differentiation has been reported from 1990s (42), it remains unclear whether intracellular Ca 2ϩ transient is required for differentiation from hMSCs into neuronal lineage. In the current study, we demonstrated that intracellular Ca 2ϩ transient peaks at day 1 post-induction ( Fig.…”

Section: Discussionmentioning

confidence: 99%

EZH2 Regulates Neuronal Differentiation of Mesenchymal Stem Cells through PIP5K1C-dependent Calcium Signaling

Chou

Chen

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

Enhancer of zeste homolog 2 (EZH2) regulates stem cells renewal, maintenance, and differentiation into different cell lineages including neuron. Changes in intracellular Ca2؉ concentration play a critical role in the differentiation of neurons. However, whether EZH2 modulates intracellular Ca 2؉ signaling in regulating neuronal differentiation from human mesenchymal stem cells (hMSCs) still remains unclear. When hMSCs were treated with a Ca 2؉ chelator or a PLC inhibitor to block IP 3 -mediated Ca 2؉ signaling, neuronal differentiation was disrupted. EZH2 bound to the promoter region of PIP5K1C to suppress its transcription in proliferating hMSCs. Interestingly, knockdown of EZH2 enhanced the expression of PIP5K1C, which in turn increased the amount of PI(4,5)P 2 , a precursor of IP 3 , and resulted in increasing the intracellular Ca 2؉ level, suggesting that EZH2 negatively regulates intracellular Ca 2؉ through suppression of PIP5K1C. Knockdown of EZH2 also enhanced hMSCs differentiation into functional neuron both in vitro and in vivo. In contrast, knockdown of PIP5K1C significantly reduced PI(4,5)P 2 contents and intracellular Ca 2؉ release in EZH2-silenced cells and resulted in the disruption of neuronal differentiation from hMSCs. Here, we provide the first evidence to demonstrate that after induction to neuronal differentiation, decreased EZH2 activates the expression of PIP5K1C to evoke intracellular Ca 2؉ signaling, which leads hMSCs to differentiate into functional neuron lineage. Activation of intracellular Ca 2؉ signaling by repressing or knocking down EZH2 might be a potential strategy to promote neuronal differentiation from hMSCs for application to neurological dysfunction diseases. Human mesenchymal stem cells (hMSCs)4 derived from bone marrow are easily obtained (1), safely expanded in vitro and are not susceptible to malignant transformation; thus, hMSCs are suitable for therapeutic applications (2). hMSCs can be induced to differentiate into multiple lineages, including bone, fat, cartilage, as well as neuron in vitro (1,(3)(4)(5). Transplanted MSCs are able to differentiate into neuronal lineage and improve the functions of central nervous system (CNS) with ischemia (6), Parkinson disease (7), Alzheimer disease (8), spinal cord injury (9, 10), and other neurodegenerative disorders (11, 12) modeled in rodents. These findings demonstrate the potential of hMSCs for cell therapy in human CNS.Spontaneous and transient elevation in intracellular Ca 2ϩ concentration during an early period is required and critical for neuronal differentiation both in vitro and in embryonic neuron development (13). However, whether intracellular Ca 2ϩ signaling is required for neuronal differentiation from hMSCs remains unclear. The expression of transient receptor potential (TRP) proteins, TRPC1 and TRPC3, is elevated to activate store-operated calcium entry (SOCE) after differentiation of H19 -7 hippocampal neuronal cells (14). It is also known that inositol 1,4,5-trisphosphate (IP 3 ) stimulates a ligand-gated chann...

show abstract

Section: Discussionmentioning

confidence: 99%

EZH2 Regulates Neuronal Differentiation of Mesenchymal Stem Cells through PIP5K1C-dependent Calcium Signaling

Chou

Chen

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Individual activity bouts are composed of multiple cycles of action potential discharge that lengthen in duration over the course of the bout (Landmesser and O'Donovan 1984a,b). Calcium entry during bouts is necessary for the proper development of neuronal phenotype and for the regulation of neurite extension in spinal motor networks (Gu and Spitzer 1995;Holliday and Spitzer 1990).…”

Section: Introductionmentioning

confidence: 99%

Episodic Bouts of Activity Accompany Recovery of Rhythmic Output By a Neuromodulator- and Activity-Deprived Adult Neural Network

Luther

Robie

Yarotsky

et al. 2003

Journal of Neurophysiology

View full text Add to dashboard Cite

. Episodic bouts of activity accompany recovery of rhythmic output by a neuromodulator-and activity-deprived adult neural network. J Neurophysiol 90: 2720 -2730, 2003. First published July 2, 2003 10.1152/jn.00370.2003. The pyloric rhythm of the stomatogastric ganglion of the crab, Cancer borealis, slows or stops when descending modulatory inputs are acutely removed. However, the rhythm spontaneously resumes after one or more days in the absence of neuromodulatory input. We recorded continuously for days to characterize quantitatively this recovery process. Activity bouts lasting 40 -900 s began several hours after removal of neuromodulatory input and were followed by stable rhythm recovery after 1-4 days. Bout duration was not related to the intervals (0.3-800 min) between bouts. During an individual bout, the frequency rapidly increased and then decreased more slowly. Photoablation of back-filled neuromodulatory terminals in the stomatogastric ganglion (STG) neuropil had no effect on activity bouts or recovery, suggesting that these processes are intrinsic to the STG neuronal network. After removal of neuromodulatory input, the phase relationships of the components of the triphasic pyloric rhythm were altered, and then over time the phase relationships moved toward their control values. Although at low pyloric rhythm frequency the phase relationships among pyloric network neurons depended on frequency, the changes in frequency during recovery did not completely account for the change in phase seen after rhythm recovery. We suggest that activity bouts represent underlying mechanisms controlling the restructuring of the pyloric network to allow resumption of an appropriate output after removal of neuromodulatory input.

show abstract

“…For washing out excess Fluo4-AM when imaging calcium activity. 5. Inside the hood, prepare the following with a p1000 micropipette for each dissection session (more than one retina can be dissected in one session).…”

Section: Dissectionmentioning

confidence: 99%

“…For example, analyzing cells from tissues removed and dissociated at different stages allows one to discern the state of specification of individual cells at different developmental stages, that is, the fate of the cells in the absence of interactions with neighboring tissues 8,[19][20][21][22][24][25][26][27][28][29][30][31][32][33] . Primary cell culture also allows the investigator to treat the culture with specific reagents and analyze the results on a single cell level 5,8,21,24,[27][28][29][30][33][34][35][36][37][38][39] . Xenopus laevis, a classic model system for the study of early neural development 19,27,29,[31][32][40][41][42] , serves as a particularly suitable system for retinal primary cell culture 10,38,[43][44][45] .…”

Section: Introductionmentioning

confidence: 99%

“…In addition to the obvious medical relevance for understanding and treating retinal disease, the development of the vertebrate retina continues to serve as an important and elegant model system for understanding neuronal cell type determination and differentiation [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16] . The neural retina consists of six discrete cell types (ganglion, amacrine, horizontal, photoreceptors, bipolar cells, and Müller glial cells) arranged in stereotypical layers, a pattern that is largely conserved among all vertebrates 12,[14][15][16][17][18] .…”

mentioning

confidence: 99%

See 1 more Smart Citation

Dissection, Culture, and Analysis of <em>Xenopus laevis</em> Embryonic Retinal Tissue

McDonough

Allen

Ng-Sui-Hing

et al. 2012

JoVE

View full text Add to dashboard Cite

The process by which the anterior region of the neural plate gives rise to the vertebrate retina continues to be a major focus of both clinical and basic research. In addition to the obvious medical relevance for understanding and treating retinal disease, the development of the vertebrate retina continues to serve as an important and elegant model system for understanding neuronal cell type determination and differentiation [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16] . The neural retina consists of six discrete cell types (ganglion, amacrine, horizontal, photoreceptors, bipolar cells, and Müller glial cells) arranged in stereotypical layers, a pattern that is largely conserved among all vertebrates 12,[14][15][16][17][18] .While studying the retina in the intact developing embryo is clearly required for understanding how this complex organ develops from a protrusion of the forebrain into a layered structure, there are many questions that benefit from employing approaches using primary cell culture of presumptive retinal cells 7,[19][20][21][22][23] . For example, analyzing cells from tissues removed and dissociated at different stages allows one to discern the state of specification of individual cells at different developmental stages, that is, the fate of the cells in the absence of interactions with neighboring tissues 8,[19][20][21][22][24][25][26][27][28][29][30][31][32][33] . Primary cell culture also allows the investigator to treat the culture with specific reagents and analyze the results on a single cell level 5,8,21,24,[27][28][29][30][33][34][35][36][37][38][39] . Xenopus laevis, a classic model system for the study of early neural development 19,27,29,[31][32][40][41][42] , serves as a particularly suitable system for retinal primary cell culture 10,38,[43][44][45] .Presumptive retinal tissue is accessible from the earliest stages of development, immediately following neural induction 25,38,43 . In addition, given that each cell in the embryo contains a supply of yolk, retinal cells can be cultured in a very simple defined media consisting of a buffered salt solution, thus removing the confounding effects of incubation or other sera-based products 10,24,[44][45] .However, the isolation of the retinal tissue from surrounding tissues and the subsequent processing is challenging. Here, we present a method for the dissection and dissociation of retinal cells in Xenopus laevis that will be used to prepare primary cell cultures that will, in turn, be analyzed for calcium activity and gene expression at the resolution of single cells. While the topic presented in this paper is the analysis of spontaneous calcium transients, the technique is broadly applicable to a wide array of research questions and approaches (Figure 1). Video LinkThe video component of this article can be found at

show abstract

Spontaneous calcium influx and its roles in differentiation of spinal neurons in culture

Cited by 134 publications

References 38 publications

EZH2 Regulates Neuronal Differentiation of Mesenchymal Stem Cells through PIP5K1C-dependent Calcium Signaling

EZH2 Regulates Neuronal Differentiation of Mesenchymal Stem Cells through PIP5K1C-dependent Calcium Signaling

Episodic Bouts of Activity Accompany Recovery of Rhythmic Output By a Neuromodulator- and Activity-Deprived Adult Neural Network

Dissection, Culture, and Analysis of <em>Xenopus laevis</em> Embryonic Retinal Tissue

Contact Info

Product

Resources

About