Abstract
This protocol describes detailed procedures for the fluorescent high-throughput screening of small molecules that induce neurogenesis in cultures of skeletal muscle cells. The detection of neurogenesis relies on a fluorescent dye, FM 1-43, which is used to study the neuronal property of depolarization-induced synaptic vesicle recycling. Thus, small molecules with neurogenesis-inducing activity in skeletal muscle cells can be rapidly identified by measuring the fluorescence intensity of the treated cells using a fluorescent microplate reader. This protocol uses murine myoblast C2C12 cells for screening, which are readily available and relatively easy to culture. Neurogenesis of PC12 cells induced by nerve growth factor is employed as a positive control for this screening. The screening time for this protocol is 8 d, which also includes the procedure to detect depolarization-induced synaptic vesicle recycling using FM 1-43.
| Original language | English |
|---|---|
| Pages (from-to) | 835-839 |
| Number of pages | 5 |
| Journal | Nature Protocols |
| Volume | 3 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - 2008 Apr 1 |
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All Science Journal Classification (ASJC) codes
- Biochemistry, Genetics and Molecular Biology(all)
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Fluorescent high-throughput screening of chemical inducers of neuronal differentiation in skeletal muscle cells. / Williams, Darren R.; Kim, Gun Hee; Lee, Myung Ryul; Shin, Injae.
In: Nature Protocols, Vol. 3, No. 5, 01.04.2008, p. 835-839.Research output: Contribution to journal › Article
TY - JOUR
T1 - Fluorescent high-throughput screening of chemical inducers of neuronal differentiation in skeletal muscle cells
AU - Williams, Darren R.
AU - Kim, Gun Hee
AU - Lee, Myung Ryul
AU - Shin, Injae
PY - 2008/4/1
Y1 - 2008/4/1
N2 - This protocol describes detailed procedures for the fluorescent high-throughput screening of small molecules that induce neurogenesis in cultures of skeletal muscle cells. The detection of neurogenesis relies on a fluorescent dye, FM 1-43, which is used to study the neuronal property of depolarization-induced synaptic vesicle recycling. Thus, small molecules with neurogenesis-inducing activity in skeletal muscle cells can be rapidly identified by measuring the fluorescence intensity of the treated cells using a fluorescent microplate reader. This protocol uses murine myoblast C2C12 cells for screening, which are readily available and relatively easy to culture. Neurogenesis of PC12 cells induced by nerve growth factor is employed as a positive control for this screening. The screening time for this protocol is 8 d, which also includes the procedure to detect depolarization-induced synaptic vesicle recycling using FM 1-43.
AB - This protocol describes detailed procedures for the fluorescent high-throughput screening of small molecules that induce neurogenesis in cultures of skeletal muscle cells. The detection of neurogenesis relies on a fluorescent dye, FM 1-43, which is used to study the neuronal property of depolarization-induced synaptic vesicle recycling. Thus, small molecules with neurogenesis-inducing activity in skeletal muscle cells can be rapidly identified by measuring the fluorescence intensity of the treated cells using a fluorescent microplate reader. This protocol uses murine myoblast C2C12 cells for screening, which are readily available and relatively easy to culture. Neurogenesis of PC12 cells induced by nerve growth factor is employed as a positive control for this screening. The screening time for this protocol is 8 d, which also includes the procedure to detect depolarization-induced synaptic vesicle recycling using FM 1-43.
UR - http://www.scopus.com/inward/record.url?scp=43149116558&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=43149116558&partnerID=8YFLogxK
U2 - 10.1038/nprot.2008.47
DO - 10.1038/nprot.2008.47
M3 - Article
C2 - 18451791
AN - SCOPUS:43149116558
VL - 3
SP - 835
EP - 839
JO - Nature Protocols
JF - Nature Protocols
SN - 1754-2189
IS - 5
ER -