Tracking Mitochondrial DNA In Situ

2016 ◽  
pp. 81-92
Author(s):  
Anna Ligasová ◽  
Karel Koberna
Keyword(s):  
Mitochondrial Dna

scholarly journals The Sorting of Mitochondrial DNA and Mitochondrial Proteins in Zygotes: Preferential Transmission of Mitochondrial DNA to the Medial Bud

1998 ◽  
Vol 142 (3) ◽  
pp. 613-623 ◽  
Author(s):  
Koji Okamoto ◽  
Philip S. Perlman ◽  
Ronald A. Butow
Keyword(s):  
Mitochondrial Dna ◽  
Fluorescent Protein ◽  
Yeast Cells ◽  
Mitochondrial Matrix ◽  
Outer Membranes ◽  
Marker Proteins ◽  
The Matrix ◽  
Green Fluorescent ◽  
Mitochondrial Reticulum

Green fluorescent protein (GFP) was used to tag proteins of the mitochondrial matrix, inner, and outer membranes to examine their sorting patterns relative to mtDNA in zygotes of synchronously mated yeast cells in ρ+ × ρ0 crosses. When transiently expressed in one of the haploid parents, each of the marker proteins distributes throughout the fused mitochondrial reticulum of the zygote before equilibration of mtDNA, although the membrane markers equilibrate slower than the matrix marker. A GFP-tagged form of Abf2p, a mtDNA binding protein required for faithful transmission of ρ+ mtDNA in vegetatively growing cells, colocalizes with mtDNA in situ. In zygotes of a ρ+ × ρ+ cross, in which there is little mixing of parental mtDNAs, Abf2p–GFP prelabeled in one parent rapidly equilibrates to most or all of the mtDNA, showing that the mtDNA compartment is accessible to exchange of proteins. In ρ+ × ρ0 crosses, mtDNA is preferentially transmitted to the medial diploid bud, whereas mitochondrial GFP marker proteins distribute throughout the zygote and the bud. In zygotes lacking Abf2p, mtDNA sorting is delayed and preferential sorting is reduced. These findings argue for the existence of a segregation apparatus that directs mtDNA to the emerging bud.

Visualization of DNA within mitochondria by osmium-ammine staining of mouse duodenal crypt cells

1992 ◽  
Vol 101 (4) ◽  
pp. 785-793
Author(s):  
D.F. Liu ◽  
M. el-Alfy ◽  
C.P. Leblond
Keyword(s):  
Mitochondrial Dna ◽  
Electron Microscope ◽  
Serial Sections ◽  
Specific Staining ◽  
Dnase Digestion ◽  
Dividing Cells ◽  
Crypt Cells ◽  
Rotary Shadowing

Previous investigators have examined mitochondrial DNA (mtDNA) in the electron microscope (EM) after extraction from mitochondria and rotary shadowing. We have observed mtDNA in situ by the osmiumammine procedure for specific staining of DNA in the EM. The procedure was modified to improve the regularity of the staining and then applied to the rapidly dividing cells present in mouse duodenal crypts. In the stained sections of these cells, 25% of the mitochondria exhibited discrete reactive filaments. The filaments, whether observed directly or in stereopairs, appeared either irregular or arranged into distinct patterns, some of which were similar to those previously described after rotary shadowing of duplicating mtDNA: namely, simple and double circular figures, displacement loops and supercoiled forms. The filaments could be traced in serial sections of the same mitochondria and, therefore, were not artifacts. Moreover, their disappearance after DNase digestion demonstrated that they were composed of DNA. It is concluded that mtDNA can be visualized by the modified osmium-ammine technique and may show patterns that can be interpreted as phases in its replication.

scholarly journals Non-isotopic in situ hybridization method for mitochondria in oncocytes.

1994 ◽  
Vol 42 (2) ◽  
pp. 273-276 ◽  
Author(s):  
V A Varma ◽  
C M Cerjan ◽  
K L Abbott ◽  
S B Hunter
Keyword(s):  
Mitochondrial Dna ◽  
In Situ Hybridization ◽  
Coding Region ◽  
Chain Reaction ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Polymerase Chain ◽  
Endogenous Biotin ◽  
Formalin Fixed

We used in situ hybridization to specifically identify mitochondria in a series of formalin-fixed, paraffin-embedded oncocytic lesions. Digoxigenin-labeled DNA probes were generated by the polymerase chain reaction (PCR), with primers designed to amplify a mitochondrion-specific 154 BP sequence within the ND4 coding region. Probes were hybridized with mitochondrial DNA under stringent conditions. Oncocytes were strongly and consistently stained, reflecting the high copy number of mitochondrial DNA within these cells. Because of the presence of endogenous biotin within mitochondria, digoxigenin is preferable to biotin as a label for detection of mitochondria.

Replication-competent human mitochondrial DNA lacking the heavy-strand promoter region

1991 ◽  
Vol 11 (3) ◽  
pp. 1631-1637
Author(s):  
C T Moraes ◽  
F Andreetta ◽  
E Bonilla ◽  
S Shanske ◽  
S DiMauro ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Promoter Region ◽  
Direct Repeat ◽  
Progressive External Ophthalmoplegia ◽  
Mitochondrial Transcription Factor ◽  
Mtdna Deletion ◽  
Heavy Strand ◽  
Light Strand

We identified two patients with progressive external ophthalmoplegia, a mitochondrial disease, who harbored a population of partially deleted mitochondrial DNA (mtDNA) with unusual properties. These molecules were deleted from mtDNA positions 548 to 4,442 and encompassed not only rRNA sequences but the heavy-strand promoter region as well. A 13-bp direct repeat was found flanking the breakpoint precisely, with the repeat at positions 535 to 547 located within the binding site for mitochondrial transcription factor 1 (mtTF1). This is the second mtDNA deletion involving a 13-bp direct repeat reported but is at least 10 times less frequent in the patient population than the former one. In situ hybridization studies showed that transcripts under the control of the light-strand promoter were abundant in muscle fibers with abnormal proliferation of mitochondria, while transcripts directed by the heavy-strand promoter, whether of genes residing inside or outside the deleted region, were not. The efficient transcription from the light-strand promoter implies that the major heavy-and light-strand promoters, although physically close, are functionally independent, confirming previous in vitro studies.

Live-Cell Probe for In Situ Single-Cell Monitoring of Mitochondrial DNA Methylation

ACS Sensors ◽  
2021 ◽  
Author(s):  
Han Zhao ◽  
Donghan Ma ◽  
Junkai Xie ◽  
Oscar Sanchez ◽  
Fang Huang ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Mitochondrial Dna ◽  
Single Cell ◽  
Live Cell ◽  
Cell Monitoring

In situ hybridization of mitochondrial DNA in the heart of a patient with Kearns-Sayre syndrome and dilatative cardiomyopathy

1992 ◽  
Vol 23 (12) ◽  
pp. 1431-1437 ◽  
Author(s):  
J. Müller-Höcker ◽  
P. Seibel ◽  
K. Schneiderbanger ◽  
Ch. Zietz ◽  
B. Obermaier-Kusser ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
In Situ Hybridization ◽  
Kearns Sayre Syndrome ◽  
Dilatative Cardiomyopathy
Sign in / Sign up

Export Citation Format

Share Document