Distribution Of Mitochondrial DNA Research Articles

High-Throughput Image-Based Quantification of Mitochondrial DNA Synthesis and Distribution.

The vast majority of cellular processes require a continuous supply of energy, the most common carrier of which is the ATP molecule. Eukaryotic cells produce most of their ATP in the mitochondria by oxidative phosphorylation. Mitochondria are unique organelles because they have their own genome that is replicated and passed on to the next generation of cells. In contrast to the nuclear genome, there are multiple copies of the mitochondrial genome in the cell. The detailed study of the mechanisms responsible for the replication, repair, and maintenance of the mitochondrial genome is essential for understanding the proper functioning of mitochondria and whole cells under both normal and disease conditions. Here, a method that allows the high-throughput quantification of the synthesis and distribution of mitochondrial DNA (mtDNA) in human cells cultured in vitro is presented. This approach is based on the immunofluorescence detection of actively synthesized DNA molecules labeled by 5-bromo-2'-deoxyuridine (BrdU) incorporation and the concurrent detection of all the mtDNA molecules with anti-DNA antibodies. Additionally, the mitochondria are visualized with specific dyes or antibodies. The culturing of cells in a multi-well format and the utilization of an automated fluorescence microscope make it easier to study the dynamics of mtDNA and the morphology of mitochondria under a variety of experimental conditions in a relatively short time.

Diversity and distribution of mitochondrial DNA in non-Austronesian-speaking Taiwanese individuals

Many studies have described the diversity of Austronesian-speaking Taiwanese people to shed more light on their origin and their connection with the “Out of Taiwan” migrations. However, the genetic relationship between the non-Austronesian-speaking groups of Taiwan and the populations of continental Asia is still unclear. Here, we studied the diversity of mtDNA in 767 non-Austronesian speakers from 16 locations in Taiwan using partial sequencing obtained from the hypervariable segment I (HVS-I) and coding regions 8,001-9,000 and 9.801–10,900 and 85 complete mtDNA genome sequences. Bayesian analysis of population structure was used to examine their relationship with over 3662 individuals representing indigenous groups of Taiwan, continental East Asia, Japan, and Island Southeast Asia. The whole analysis identified 278 haplotypes. Complete genomes revealed 62 novel subhaplogroups, of which 31 were exclusive to Taiwan. Estimates of coalescence times of all subhaplogroups showed peaks of diversification greater than 5.0 kya, likely characterizing gene flow from continental East Asian groups but not excluding in situ Taiwanese ancestry. Furthermore, a significant number of clades exclusive to non-Austronesian speakers of Taiwan (NAN_Tw) showed coalescence peaks between 1.0 and 2.6 kya, suggesting possible late Neolithic to early metal age settlements of NAN_Tw and local expansion in Taiwan.

Mitochondrial dynamics in health and disease.

In animals, mitochondria are mainly organised into an interconnected tubular network extending across the cell along a cytoskeletal scaffold. Mitochondrial fission and fusion, as well as distribution along cytoskeletal tracks, are counterbalancing mechanisms acting in concert to maintain a mitochondrial network tuned to cellular function. Balanced mitochondrial dynamics permits quality control of the network including biogenesis and turnover, and distribution of mitochondrial DNA, and is linked to metabolic status. Cellular and organismal health relies on a delicate balance between fission and fusion, and large rearrangements in the mitochondrial network can be seen in response to cellular insults and disease. Indeed, dysfunction in the major components of the fission and fusion machineries including dynamin-related protein 1 (DRP1), mitofusins 1 and 2 (MFN1, MFN2) and optic atrophy protein 1 (OPA1) and ensuing imbalance of mitochondrial dynamics can lead to neurodegenerative disease. Altered mitochondrial dynamics is also seen in more common diseases. In this review, the machinery involved in mitochondrial dynamics and their dysfunction in disease will be discussed.

OPA1-Exon4b Binds to mtDNA D-Loop for Transcriptional and Metabolic Modulation, Independent of Mitochondrial Fusion

Optic Atrophy 1 (OPA1) has well-established roles in both mitochondrial fusion and apoptotic crista remodeling and is required for the maintenance and distribution of mitochondrial DNA (mtDNA), which are essential for energy metabolism. However, the relationship between OPA1 and mitochondrial metabolism and the underlying mechanisms remain unclear. Here, we show that OPA1-Exon4b modulates mitochondrial respiration and rescues inner mitochondrial membrane potential (Δψm), independent of mitochondrial fusion. OPA1-Exon4b is required for the maintenance of normal TFAM distribution and enhances mtDNA transcription by binding the D-loop of mtDNA. Finally, we show that mRNA levels of OPA1 isoforms containing Exon4b are specifically downregulated in hepatocellular carcinoma (HCC), leading to a reduction in Δψm. Thus, our study demonstrates a novel mitochondrial functional self-recovery pathway involving enhanced mtDNA transcription-mediated recovery of mitochondrial respiratory chain proteins. This mitochondrial fusion-independent pathway may contribute to mitochondrial multi-functional switches in tumorigenesis.

MtDNA structure: the women who formed the Brazilian Northeast

BackgroundThe distribution of mitochondrial DNA (mtDNA) lineages in Brazil is heterogeneous due to different regional colonization dynamics. Northeastern Brazil, although being an important region in terms of human imigration and ethnic admixture, has little information regarding its population mtDNA composition. Here, we determine which mitochondrial lineages contributed to the formation of the Northeastern Brazilian population. Our sample consisted of 767 individuals distributed as follows i) 550 individuals from eight Northeastern states (Piauí, Ceará, Rio Grande do Norte, Paraíba, Pernambuco, Alagoas, Sergipe, and Bahia) which were sequenced for mtDNA hypervariable segments I, II, and III; ii) 217 individuals from Alagoas and Pernambuco (previously published data). Data analysis was performed through sequence alignment and Haplogrep 2.0 haplogroup assignment tools. Furthermore, maternal ancestry distribution was contextualized and, when possible, related to historical events to better understand the biological interactions and population dynamics that occurred in this region since the beginning of colonization.ResultsUnexpectedly, Amerindian mitochondrial ancestry was the highest in the Northeastern region overall, followed by African, European and non-Amerindian Asian, unlike previous results for this region. Alagoas and Pernambuco states, however, showed a larger African mtDNA frequency. The Northeastern region showed an intraregional heterogeneous distribution regarding ancestral groups, in which states/mesoregions located to the north had a prevalent Amerindian ancestral frequency and those to the south had predominance of African ancestry. Moreover, results showed great diversity of European haplogroups and the presence of non-Amerindian Asian haplogroups.ConclusionsOur findings are in disagreement with previous investigations that suggest African mitochondrial ancestry is the most prevalent in the Brazilian Northeast. The predominance of Amerindian lineages exemplifies the importance of indigenous women in the formation of the population, despite intense African slave entry and conflicts with European settlers. The variable distribution of ancestral groups observed in the Northeast is in accordance with historical records showing the similarities with colonization dynamics occurred in the Amazon region and the Brazilian Southeast. Moreover, the variety of European haplogroups suggests multiple origins of founding groups, specially those found in Western European populations.

A Difference in the Genetic Distribution Pattern between The Sexes in the Asian Black Bear

Abstract The genetic structures of animal populations are influenced by the difference in the dispersal patterns of the sexes. To reveal effects caused by different dispersal systems between sexes on genetic structure, we examined the differences in the distribution of mitochondrial DNA in Asian black bears and compared the relationship between genetic correlation coefficients and pairwise distances of the bears between sexes using microsatellite DNA. From sequences of 758 black bears from continuous populations, we examined sequence data for 326 samples carrying ten haplotypes that were distributed locally. The estimated pairwise rectilinear distances indicated that males tended to distribute around the inner edge of the observed area of the locally distributed haplotypes. Distribution range analysis showed that both the maximum and average ranges of males in eight locally distributed haplotypes were larger than those of females. We also performed spatial autocorrelation analysis using genotype data for ...

Lineage distribution and barriers to gene flow among populations of the globally invasive marine mussel Musculista senhousia

Introduced species seem to rapidly adapt to non-native conditions although founder events may decrease genetic variance and impede adaptation. Cryptic introductions of an invader from different parts of its native range may allow colonization of habitat patches with compatible conditions, or admixture may restore genetic diversity lost during invasion bottlenecks. We present the first genetic study of global invasion pathways for the Asian date mussel, Musculista senhousia, comparing three putative source populations from Asia with six US estuaries and one invaded site in New Zealand and the Mediterranean Sea, totaling 117 individuals. Distribution of mitochondrial DNA and alleles of a nuclear intron revealed two genetically divergent populations in the native range. A warm-water lineage from Tokyo Bay and southwestern South Korea predominated in southern California and New Zealand, while a cold-water haplotype group sampled in northeastern Korea was more abundant at invaded sites in the northwestern US and Mediterranean Sea. The duration of high summer water temperatures accounted for substantial variance in the worldwide distribution of common haplotype groups, suggesting thermal pre-adaptation may influence invasion success for differentiated source populations. There was no gene flow across central California, but on either side of this phylogeographic barrier there was high exchange among estuaries within the northern and the southern US regional populations. Gradients in water temperature may therefore constrain post-introduction spread by limiting dispersion along a coast.

OPA1 links human mitochondrial genome maintenance to mtDNA replication and distribution

Eukaryotic cells harbor a small multiploid mitochondrial genome, organized in nucleoids spread within the mitochondrial network. Maintenance and distribution of mitochondrial DNA (mtDNA) are essential for energy metabolism, mitochondrial lineage in primordial germ cells, and to prevent mtDNA instability, which leads to many debilitating human diseases. Mounting evidence suggests that the actors of the mitochondrial network dynamics, among which is the intramitochondrial dynamin OPA1, might be involved in these processes. Here, using siRNAs specific to OPA1 alternate spliced exons, we evidenced that silencing of the OPA1 variants including exon 4b leads to mtDNA depletion, secondary to inhibition of mtDNA replication, and to marked alteration of mtDNA distribution in nucleoid and nucleoid distribution throughout the mitochondrial network. We demonstrate that a small hydrophobic 10-kDa peptide generated by cleavage of the OPA1-exon4b isoform is responsible for this process and show that this peptide is embedded in the inner membrane and colocalizes and coimmunoprecipitates with nucleoid components. We propose a novel synthetic model in which a peptide, including two trans-membrane domains derived from the N terminus of the OPA1-exon4b isoform in vertebrates or from its ortholog in lower eukaryotes, might contribute to nucleoid attachment to the inner mitochondrial membrane and promotes mtDNA replication and distribution. Thus, this study places OPA1 as a direct actor in the maintenance of mitochondrial genome integrity.

Cell cycle dependent morphology changes and associated mitochondrial DNA redistribution in mitochondria of human cell lines

Mitochondria of osteosarcoma cells (143B) in culture have variable morphologies, classified according to the shape and size of the organelle as reticular, fragmented or intermediate. Synchronization and release from G0 has shown that the morphology of mitochondria oscillates between the reticular and fragmented state in a cell cycle dependent manner. Cells in G1 have reticular mitochondria while those in S phase have fragmented mitochondria. By using a novel method of fluorescence in situ hybridization, the morphology of mitochondria was correlated with mitochondrial DNA distribution. MtDNA molecules were seen in clusters of two to four along mitochondrial filaments. In the fully fragmented state, each mitochondrion contained at least one cluster. We discuss the importance of fission and fusion events in regulating the morphology of mitochondria, segregation of mtDNA and maintenance of the organelle's functional unity.

Extreme population structuring in the threatened ghost bat, Macroderma gigas: evidence from mitochondrial DNA.

The ghost bat, Macroderma gigas, has undergone a major range contraction and is currently restricted around a few, highly disjunct maternity sites. The amount and distribution of mitochondrial DNA (mtDNA) variation within extant populations has been used to assess levels of current and historical maternal gene flow between these populations. An approximately 330 base pair fragment spanning a hypervariable area of the mtDNA control region was amplified and sequenced by using 22 individuals from four current ghost bat populations. The mean sequence diversity of 4.5% between populations was six times higher than that within populations (0.68%), and alleles within populations were monophyletic. Restriction enzyme analysis of amplified products from an additional 100 individuals revealed fixed allelic differences in the distribution of control region genotypes between the four populations. It is suggested that this extreme genetic subdivision is a consequence of long-term female philopatry. For the purposes of management each population should be treated as an independent entity. The depth of the genetic structuring suggests that the isolation of extant populations preceded the historical range contraction.

Studies on Mitochondrial Structure and Function in Physarum polycephalum. VI. Synthesis and Distribution of Mitochondrial DNA as Revealed by Light Microscope Autoradiography

The synthesis and distribution of mitochondrial DNA (mt-DNA) were studied by light microscope autoradiography in the plasmodium of Physarum polycephalum. Mitochondria pre-labeled with 3H-thymidine (3H-TdR) for 3 h during early mitochondrial S phase (mS) and incubated in non-radioactive medium showed elongated shapes that developed to dumbbell-shaped, and the latter form of mitochondria divided. The incorporation of 3H-TdR into mitochondria occurred during mitochondrial growth but terminated before development to dumbbell-shapes, suggesting that they synthesized mtDNA during a limited period of mitochondrial division cycle. In addition, mitochondrial grains counts were reduced remarkably to about half during the late nuclear S phase, suggesting that Physarum mitochondria divided semi-synchronously during late nuclear S phase. The distribution of mtDNA pre-labeled during early or late mS was determined by the grain distribution of (a) labeling of both poles of dumbbell-shaped mitochondria, and (b) labeling of daughter mitochondria. The results indicated that mtDNA was distributed equally to daughter mitochondria, regardless of time of mtDNA replication.

Segregation of mitochondrially inherited antibiotic resistance genes in zygote cell lineages of Saccharomyces cerevisiae

An analysis of cell lineages from individual zygotes derived from crosses involving two mitochondrial antibiotic resistance genes suggests that The segregation and recombination of mitochondrial genes is discussed in relation to the possile distribution of mitochondrial DNA during budding of zygotes and zygote daughters.