Abstract:
BACKGROUND:With advances in cell capture, gene expression can now be studied in neuronal subtypes and single cells; however, studying epigenetic mechanisms that underlie these changes presents challenges. Moreover, chromatin immunoprecipitation (ChIP) protocols optimized for low cell number do not adequately address technical issues and cell loss while preparing tissue for fluorescence activated cell sorting (FACS). Developing a reliable FACS-ChIP protocol without the need for pooling tissue from multiple animals would enable study of epigenetic mechanisms in neuronal subtypes. METHODS:FACS was used to isolate dopamine 1 receptor (D1R) expressing cells from the nucleus accumbens (NAc) of a commercially available BAC transgenic mouse strain. D1R+ cells were used to study gene expression as well as histone modifications at gene promoters using a novel native ChIP protocol. RESULTS:Isolated cells had enrichment of the dopamine 1 receptor (D1R) mRNA and nearly undetectable levels of GFAP and D2R mRNA. ChIP analysis demonstrated the association of activating or repressive histone modifications with highly expressed or silent gene promoters, respectively. COMPARISON WITH EXISTING METHODS:The ChIP protocol developed in this paper enables characterization of histone modifications from ∼30,000 FAC-sorted neurons. CONCLUSIONS:We describe a one day FACS-ChIP protocol that can be applied to epigenetic studies of neuronal subtypes without pooling tissue.
journal_name
J Neurosci Methodsjournal_title
Journal of neuroscience methodsauthors
Finegersh A,Homanics GEdoi
10.1016/j.jneumeth.2016.02.006subject
Has Abstractpub_date
2016-04-01 00:00:00pages
81-8eissn
0165-0270issn
1872-678Xpii
S0165-0270(16)00058-3journal_volume
263pub_type
杂志文章abstract::Bone marrow stroma cells-derived neural stem cells (BMSCs-D-NSCs) transplantation is a promising strategy for the treatment of nervous system disorders. The development of a non-invasive method to follow the fate of BMSCs-D-NSCs in vivo is very important for the future application of this treatment. In this paper, we ...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/j.jneumeth.2009.01.007
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abstract::We describe a much simplified high-performance liquid chromatography (HPLC) method for the measurement of caffeine in plasma and brain. A particularly attractive feature of this method is that a simple methanol/water (60:40) mobile phase can be used both for plasma and brain samples. In addition, the method is compati...
journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
pub_type: 杂志文章,评审
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/j.jneumeth.2008.07.015
更新日期:2008-09-30 00:00:00
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
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更新日期:2016-01-30 00:00:00
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journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/0165-0270(94)90208-9
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journal_title:Journal of neuroscience methods
pub_type: 杂志文章
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journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/j.jneumeth.2019.108400
更新日期:2019-11-01 00:00:00
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
pub_type: 杂志文章
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更新日期:1988-09-01 00:00:00
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journal_title:Journal of neuroscience methods
pub_type: 杂志文章
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