A novel reporter of notch signalling indicates regulated and random notch activation during vertebrate neurogenesis View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2011-08-31

AUTHORS

Filipe Vilas-Boas, Rita Fior, Jason R Swedlow, Kate G Storey, Domingos Henrique

ABSTRACT

BackgroundBuilding the complex vertebrate nervous system involves the regulated production of neurons and glia while maintaining a progenitor cell population. Neurogenesis starts asynchronously in different regions of the embryo and occurs over a long period of time, allowing progenitor cells to be exposed to multiple extrinsic signals that regulate the production of different cell types. Notch-mediated cell-cell signalling is one of the mechanisms that maintain the progenitor pool, however, little is known about how the timing of Notch activation is related to the cell cycle and the distinct modes of cell division that generate neurons. An essential tool with which to investigate the role of Notch signalling on cell by cell basis is the development a faithful reporter of Notch activity.ResultsHere we present a novel reporter for Notch activity based on the promoter of the well characterised Notch target chick Hes5-1, coupled with multiple elements that confer instability, including a destabilized nuclear Venus fluorescent protein and the 3' untranslated region (UTR) of Hes5-1. We demonstrate that this reporter faithfully recapitulates the endogenous expression of Hes5-1 and that it robustly responds to Notch activation in the chick neural tube. Analysis of the patterns of Notch activity revealed by this reporter indicates that although Notch is most frequently activated prior to mitosis it can be activated at any time within the cell cycle. Notch active progenitors undergoing mitosis generate two daughters that both continue to experience Notch signalling. However, cells lacking Notch activity before and during mitosis generate daughters with dissimilar Notch activity profiles.ConclusionsA novel Notch reporter with multiple destabilisation elements provides a faithful read-out of endogenous Notch activity on a cell-by-cell basis, as neural progenitors progress through the cell cycle in the chick neural tube. Notch activity patterns in this cell population provide evidence for distinct Notch signalling dynamics underlying different cell division modes and for the involvement of random initiation of Notch signalling within the neuroepithelium. These findings highlight the importance of single-cell analysis in the study of the complexity of Notch activity and provide new insights into the mechanisms underlying cell fate decisions in neural progenitors. More... »

PAGES

58

Identifiers

URI

http://scigraph.springernature.com/pub.10.1186/1741-7007-9-58

DOI

http://dx.doi.org/10.1186/1741-7007-9-58

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1025117342

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/21880129


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40 schema:description BackgroundBuilding the complex vertebrate nervous system involves the regulated production of neurons and glia while maintaining a progenitor cell population. Neurogenesis starts asynchronously in different regions of the embryo and occurs over a long period of time, allowing progenitor cells to be exposed to multiple extrinsic signals that regulate the production of different cell types. Notch-mediated cell-cell signalling is one of the mechanisms that maintain the progenitor pool, however, little is known about how the timing of Notch activation is related to the cell cycle and the distinct modes of cell division that generate neurons. An essential tool with which to investigate the role of Notch signalling on cell by cell basis is the development a faithful reporter of Notch activity.ResultsHere we present a novel reporter for Notch activity based on the promoter of the well characterised Notch target chick Hes5-1, coupled with multiple elements that confer instability, including a destabilized nuclear Venus fluorescent protein and the 3' untranslated region (UTR) of Hes5-1. We demonstrate that this reporter faithfully recapitulates the endogenous expression of Hes5-1 and that it robustly responds to Notch activation in the chick neural tube. Analysis of the patterns of Notch activity revealed by this reporter indicates that although Notch is most frequently activated prior to mitosis it can be activated at any time within the cell cycle. Notch active progenitors undergoing mitosis generate two daughters that both continue to experience Notch signalling. However, cells lacking Notch activity before and during mitosis generate daughters with dissimilar Notch activity profiles.ConclusionsA novel Notch reporter with multiple destabilisation elements provides a faithful read-out of endogenous Notch activity on a cell-by-cell basis, as neural progenitors progress through the cell cycle in the chick neural tube. Notch activity patterns in this cell population provide evidence for distinct Notch signalling dynamics underlying different cell division modes and for the involvement of random initiation of Notch signalling within the neuroepithelium. These findings highlight the importance of single-cell analysis in the study of the complexity of Notch activity and provide new insights into the mechanisms underlying cell fate decisions in neural progenitors.
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46 schema:keywords Notch
47 Notch activation
48 Notch activity
49 Notch reporter
50 ResultsHere
51 Venus fluorescent protein
52 activation
53 active progenitors
54 activity
55 activity patterns
56 activity profiles
57 analysis
58 basis
59 cell basis
60 cell cycle
61 cell division
62 cell division modes
63 cell fate decisions
64 cell populations
65 cell types
66 cell-cell signaling
67 cells
68 chick neural tube
69 complexity
70 cycle
71 daughters
72 decisions
73 development
74 different cell types
75 different regions
76 distinct modes
77 distinct notch
78 division
79 division mode
80 dynamics
81 elements
82 embryos
83 endogenous expression
84 essential tool
85 evidence
86 expression
87 extrinsic signals
88 faithful reporter
89 fate decisions
90 findings
91 fluorescent protein
92 generate neurons
93 glia
94 importance
95 initiation
96 insights
97 instability
98 involvement
99 long period
100 mechanism
101 mitosis
102 mode
103 multiple elements
104 multiple extrinsic signals
105 nervous system
106 neural progenitors
107 neural tube
108 neuroepithelium
109 neurogenesis
110 neurons
111 new insights
112 novel reporter
113 patterns
114 period
115 pool
116 population
117 production
118 profile
119 progenitor cell populations
120 progenitor cells
121 progenitor pool
122 progenitors
123 promoter
124 protein
125 random initiation
126 region
127 regulated production
128 reporter
129 role
130 role of Notch
131 signaling
132 signals
133 single-cell analysis
134 study
135 system
136 time
137 timing
138 tool
139 tube
140 types
141 untranslated region
142 vertebrate neurogenesis
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