Abstract：
:Mitochondrial DNA (mtDNA) replication and transcription are crucial for cell function, but these processes are poorly understood at the single-cell level. We describe a novel fluorescence in situ hybridization protocol, called mTRIP (mitochondrial transcription and replication imaging protocol), that reveals simultaneously mtDNA and RNA, and that can also be coupled to immunofluorescence for in situ protein examination. mTRIP reveals mitochondrial structures engaged in initiation of DNA replication by identification of a specific sequence in the regulatory D-loop, as well as unique transcription profiles in single human cells. We observe and quantify at least three classes of mitochondrial structures: (i) replication initiation active and transcript-positive (Ia-Tp); (ii) replication initiation silent and transcript-positive (Is-Tp); and (iii) replication initiation silent and transcript-negative (Is-Tn). Thus, individual mitochondria are dramatically heterogeneous within the same cell. Moreover, mTRIP exposes a mosaic of distinct nucleic acid patterns in the D-loop, including H-strand versus L-strand transcripts, and uncoupled rRNA transcription and mtDNA initiation of replication, which might have functional consequences in the regulation of the mtDNA. Finally, mTRIP identifies altered mtDNA processing in cells with unbalanced mtDNA content and function, including in human mitochondrial disorders. Thus, mTRIP reveals qualitative and quantitative alterations that provide additional tools for elucidating the dynamics of mtDNA processing in single cells and mitochondrial dysfunction in diseases.
journal_name
J Cell Scijournal_title
Journal of cell scienceauthors
Chatre L,Ricchetti Mdoi
10.1242/jcs.114322subject
Has Abstractpub_date
2013-02-15 00:00:00pages
914-26issue
Pt 4eissn
0021-9533issn
1477-9137pii
jcs.114322journal_volume
126pub_type
杂志文章abstract：:The morphogenesis-related NDR kinase (MOR) pathway regulates morphogenesis in fungi. In spite of the high conservation of its components, impairing their functions results in highly divergent cellular responses depending on the fungal species. The reasons for such differences are unclear. Here we propose that the spec...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：10.1242/jcs.107862
更新日期：2012-10-01 00:00:00
abstract：:The constitutive (or default) pathway for protein secretion was investigated in two epithelial cells, Madin-Darby canine kidney (MDCK) and human colonic adenocarcinoma (Caco-2), using a bacterial enzyme. The choice of a bacterial protein was based on the requirement to identify a protein devoid of sorting signals. The...
journal_title：Journal of cell science
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doi：
更新日期：1992-07-01 00:00:00
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journal_title：Journal of cell science
pub_type： 杂志文章
doi：
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journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：1983-09-01 00:00:00
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journal_title：Journal of cell science
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journal_title：Journal of cell science
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doi：
更新日期：1999-04-01 00:00:00
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journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：1990-09-01 00:00:00
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journal_title：Journal of cell science
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更新日期：2017-07-15 00:00:00
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journal_title：Journal of cell science
pub_type： 杂志文章,评审
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更新日期：2014-12-15 00:00:00
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journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：1977-04-01 00:00:00
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journal_title：Journal of cell science
pub_type： 杂志文章,评审
doi：
更新日期：2000-12-01 00:00:00
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pub_type： 杂志文章
doi：10.1242/jcs.011130
更新日期：2007-11-01 00:00:00
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journal_title：Journal of cell science
pub_type： 杂志文章
doi：10.1242/jcs.128009
更新日期：2014-10-01 00:00:00
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journal_title：Journal of cell science
pub_type： 杂志文章
doi：10.1242/jcs.01398
更新日期：2004-11-01 00:00:00
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journal_title：Journal of cell science
pub_type： 杂志文章,评审
doi：
更新日期：2000-01-01 00:00:00
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journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：2001-08-01 00:00:00
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journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：1980-06-01 00:00:00
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journal_title：Journal of cell science
pub_type： 杂志文章
doi：10.1242/jcs.116533
更新日期：2012-12-15 00:00:00
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journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：1986-07-01 00:00:00
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journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：2000-02-01 00:00:00
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journal_title：Journal of cell science
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更新日期：2020-07-16 00:00:00
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journal_title：Journal of cell science
pub_type： 杂志文章
doi：
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journal_title：Journal of cell science
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doi：
更新日期：1998-11-01 00:00:00
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journal_title：Journal of cell science
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更新日期：1997-01-01 00:00:00
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更新日期：2005-02-15 00:00:00
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journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：1999-07-01 00:00:00
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更新日期：2012-07-01 00:00:00
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pub_type： 杂志文章
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