| Detection of histone modifications at specific gene loci in single cells in histological sections. | |
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MedLine Citation:
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PMID: 23314172 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Chromatin immunoprecipitation assays have contributed greatly to our understanding of the role of histone modifications in gene regulation. However, they do not permit analysis with single-cell resolution, thus confounding analyses of heterogeneous cell populations. Here we present a method that permits visualization of histone modifications of single genomic loci with single-cell resolution in formaldehyde-fixed paraffin-embedded tissue sections based on combined use of in situ hybridization and proximity ligation assays. We show that dimethylation of lysine 4 of histone H3 (H3K4me2) at the MYH11 locus is restricted to the smooth muscle cell (SMC) lineage in human and mouse tissue sections and that the mark persists even in phenotypically modulated SMC in atherosclerotic lesions that show no detectable expression of SMC marker genes. This methodology has promise for broad applications in the study of epigenetic mechanisms in complex multicellular tissues in development and disease. |
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Authors:
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Delphine Gomez; Laura S Shankman; Anh T Nguyen; Gary K Owens |
Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Validation Studies Date: 2013-01-13 |
Journal Detail:
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Title: Nature methods Volume: 10 ISSN: 1548-7105 ISO Abbreviation: Nat. Methods Publication Date: 2013 Feb |
Date Detail:
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Created Date: 2013-01-30 Completed Date: 2013-03-28 Revised Date: 2013-04-16 |
Medline Journal Info:
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Nlm Unique ID: 101215604 Medline TA: Nat Methods Country: United States |
Other Details:
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Languages: eng Pagination: 171-7 Citation Subset: IM |
Affiliation:
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Robert M. Berne Cardiovascular Research Center, University of Virginia School of Medicine, Charlottesville, Virginia, USA. |
| Data Bank Information | |
Bank Name/Acc. No.:
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RefSeq/NG_009299; NM_002474; NM_013607 |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Atherosclerosis / metabolism Cell Lineage Chromatin Immunoprecipitation Epigenesis, Genetic Fixatives Formaldehyde Histones / metabolism* Humans In Situ Hybridization Lysine / metabolism* Male Methylation Mice Myocytes, Smooth Muscle / metabolism Myosin Heavy Chains / metabolism Paraffin Embedding |
| Grant Support | |
ID/Acronym/Agency:
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R01 HL038854/HL/NHLBI NIH HHS; R01 HL057353/HL/NHLBI NIH HHS; R01 HL087867/HL/NHLBI NIH HHS; R01 HL087867/HL/NHLBI NIH HHS; R01 HL098538/HL/NHLBI NIH HHS; R01 HL098538/HL/NHLBI NIH HHS; R01 HL57353/HL/NHLBI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Fixatives; 0/Histones; 0/MYH11 protein, human; 0/Myosin Heavy Chains; 0/myosin 11, mouse; 50-00-0/Formaldehyde; 56-87-1/Lysine |
| Comments/Corrections | |
Comment In:
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Nat Methods. 2013 Feb;10(2):117-8
[PMID:
23361091
]
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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