Quantification of mitochondrial DNA in peripheral blood mononuclear cells and subcutaneous fat using real-time polymerase chain reaction

Gahan, Michelle E.; Miller, Francis; Lewin, Sharon R.; Cherry, Catherine L.; Hoy, Jennifer; Mijch, Anne M; Rosenfeldt, Franklin; Wesselingh, Steven Lodewyk

doi:10.1016/s1386-6532(01)00195-0

Cited by 68 publications

(28 citation statements)

References 13 publications

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“…Furthermore, treatment of diabetic individuals with pioglitazone restored the reduced level of mitochondrial number to that observed in nondiabetic control subjects. It has been previously reported that the real-time PCR method used in our experiment is highly sensitive to determine the mitochondrial copy number in several tissue including adipocytes (23,27). We found that the increase in mitochondrial DNA was mirrored by an increase in the citrate synthase activity, a mitochondrial protein encoded by a nuclear gene that is widely recognized as a marker of mitochondrial activity and correlated to mitochondrial number in human muscle (28).…”

Section: Discussionsupporting

confidence: 50%

Pioglitazone Induces Mitochondrial Biogenesis in Human Subcutaneous Adipose Tissue In Vivo

et al. 2005

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Section: Discussionsupporting

confidence: 50%

Pioglitazone Induces Mitochondrial Biogenesis in Human Subcutaneous Adipose Tissue In Vivo

et al. 2005

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“…We found that the average number of mtDNA copies in adipose tissue was about 1,000 per cell. Mean mtDNA copy number per cell in adipose tissue has previously been reported to be from 500 to 2,000 [12,24]. We believe the different results are primarily due to experimental variation due to different efficiencies in amplifying plasmids used for constructing standard curves, use of different DNA extracting kits and PCR machines, as well as pipetting errors.…”

Section: Discussionmentioning

confidence: 94%

Strong association between mitochondrial DNA copy number and lipogenesis in human white adipose tissue

et al. 2007

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Aims/hypothesis Recent studies suggest a link between insulin resistance and mitochondrial function in white fat cells. The aim of this study was to evaluate adipocyte mitochondrial DNA (mtDNA) copy number in relation to adipocyte and clinical variables that are related to insulin sensitivity. Methods We studied a group of 148 healthy volunteers with a large inter-individual variation in BMI. Relative amounts of mtDNA and nuclear DNA were determined by quantitative RT-PCR. The mtDNA:nuclear DNA ratio reflects the tissue concentration of mtDNA per cell. Results The mtDNA copy number was enriched in adipocytes of adipose tissue and decreased slightly by ageing (p= 0.015) and increasing BMI (p=0.004); however, it was not influenced by sex, energy-restricted diets or marked longterm weight reduction. Adipose mtDNA copy number was not independently related to resting energy expenditure, overall insulin sensitivity or adipocyte lipolysis. However, it showed a strong positive correlation with basal (p=0.0012) and insulin-stimulated lipogenesis (p<0.0001) in fat cells, independently of age and BMI, and a weak positive correlation with levels of mRNA from several genes involved in mitochondrial oxidative capacity (r=0.2-0.3). Conclusions/interpretationThe mtDNA copy number in human white fat cells is fairly stable within healthy individuals. It is not influenced by sex or weight loss and is not important for overall insulin sensitivity or energy expenditure at rest. However, it is strongly related to adipocyte lipogenesis and weakly to mitochondrial oxidative capacity, suggesting that adipocyte mitochondria are, above all, local regulators.

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“…significant differences in mtDNA copy number between, for example, skeletal and cardiac muscle (Miller et al 2003) and peripheral blood mononuclear cells and subcutaneous fat (Gahan et al 2001). However, for the generation of homoplasmic SCNT embryos, the choice of donor cell may undoubtedly affect the level of DmtDNA transmitted.…”

Section: Tablementioning

confidence: 99%

Aberrant Nucleo-cytoplasmic Cross-Talk Results in Donor Cell mtDNA Persistence in Cloned Embryos

et al. 2006

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Mitochondrial DNA is an extranuclear genome normally maternally inherited through the oocyte. However, the use of nuclear transfer can result in both donor cell and recipient oocyte mitochondrial DNA persisting through to blastocyst and being transmitted to the offspring. The degree of donor mitochondrial DNA transmission appears to be random and currently no evidence exists to explain this phenomenon. To determine whether this is a dilution factor or directly related to the transcriptional status of the donor cell in respect of mitochondrial DNA transcription factors, we have generated sheep nuclear transfer embryos using donor cells: (1) possessing their full mitochondrial DNA complement, (2) those partially depleted, and (3) those depleted but containing residual levels. For each donor type, donor mitochondrial DNA persisted in some blastocysts. It is evident from the donor cells used that nuclearencoded mitochondrial DNA transcription and replication factors persist even after mitochondrial DNA depletion, as do transcripts for some of the mitochondrial-encoded genes. These cells are therefore still programmed to drive mitochondrial DNA replication and transcription. In nuclear transfer-derived embryos, we have observed the persistence of these nuclear-encoded mitochondrial DNA transcription and replication factors but not in those embryos generated through in vitro fertilization. Consequently, nucleo-mitochondrial interaction following nuclear transfer is out of sequence as the onset of mitochondrial replication is a postimplantation event.

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Quantification of mitochondrial DNA in peripheral blood mononuclear cells and subcutaneous fat using real-time polymerase chain reaction

Cited by 68 publications

References 13 publications

Pioglitazone Induces Mitochondrial Biogenesis in Human Subcutaneous Adipose Tissue In Vivo

Pioglitazone Induces Mitochondrial Biogenesis in Human Subcutaneous Adipose Tissue In Vivo

Strong association between mitochondrial DNA copy number and lipogenesis in human white adipose tissue

Aberrant Nucleo-cytoplasmic Cross-Talk Results in Donor Cell mtDNA Persistence in Cloned Embryos

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