Concurrent Measurement of Mitochondrial DNA Copy Number and ATP Concentration in Single Bovine Oocytes

Read, Casey C; Bhandari, Sadikshya; Moorey, Sarah E.

doi:10.3390/mps4040088

Cited by 6 publications

(3 citation statements)

References 31 publications

(48 reference statements)

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“…In our study, the number and proportion of individual mitochondria in the oocytes did not differ between the two age groups. In a study involving a different quantification method (mitochondrial DNA) [ 7 ], a decline in the number of mitochondria with advancing age was reported; however, there is a lack of correlation between the mitochondrial DNA copy numbers and ATP content of oocytes, presumably due to ATP generation by a subpopulation of mitochondria within an oocyte [ 31 ]. Given the dynamic nature of the mitochondrial population, capable of fusing, dividing, and moving to meet cellular energy demands, a crucial quality control mechanism like mitophagy becomes essential for rectifying damaged mitochondria.…”

Section: Discussionmentioning

confidence: 99%

Effect of maternal age on ATP content and distribution of mitochondria in bovine oocytes

Dadarwal,

Pfeifer,

Cervantes

et al. 2024

PLoS ONE

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Our objective was to understand how maternal age influences the mitochondrial population and ATP content of in vivo matured bovine oocytes. We hypothesized that in vivo matured oocytes from older cows would have altered mitochondrial number and distribution patterns and lower cytoplasmic ATP content compared to the oocytes obtained from younger cows. Follicles ≥5mm were ablated in old cows (13 to 22 yrs, Old Group, n = 7) and their younger daughters (4 to 10 years old, Young Group; n = 7) to induce the emergence of a new follicular wave. Cows were treated twice daily with eight doses of FSH starting 24 hr after ablation (Day 0, day of wave emergence). Prostaglandin F2alpha (PGF) was given on Days 3 and 3.5, LH on Day 4.5, and cumulus-oocyte-complexes were collected 18–20 hours post-LH by ultrasound-guided follicular aspiration. Oocytes were either processed for staining with MitoTracker Deep Red FM or for ATP assay. Stained oocytes were imaged with a Zeiss LSM 710 confocal microscope, and mitochondria were segmented in the oocyte volume sets using Imaris Pro 7.4. In vivo matured oocytes obtained from old cows were similar in morphological grades to those from young cows. However, the oocytes of COC from older cows had 23% less intracellular ATP (27.4±1.9 vs 35.7±2.2 pmol per oocyte, P = 0.01) than those of young cows. Furthermore, the average volume of individual mitochondria, indicated by the number of image voxels, was greater (P<0.05) in oocytes from older cows than in those from younger cows. Oocytes from older cows also tended to have a greater number of mitochondrial clusters (P = 0.06) and an increased number of clusters in the central region of the oocytes (P = 0.04) compared to those from younger cows. In conclusion, our study demonstrated that maternal age was associated with a decrease in the cytoplasmic ATP content of in vivo mature oocytes and an altered distribution of mitochondrial structures. These findings suggest that maternal age may negatively influence the developmental competence of oocytes from older cows.

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Section: Discussionmentioning

confidence: 99%

Effect of maternal age on ATP content and distribution of mitochondria in bovine oocytes

Dadarwal,

Pfeifer,

Cervantes

et al. 2024

PLoS ONE

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“…4 C). Furthermore, the number of mitochondria gradually increases, thus suggesting that the active functional state at this stage requires more ATP from the mitochondria [ 92 ]. During spermatogenesis, Mfns exhibit dynamic expression during different stages of spermatogenesis; the expression levels of Mfn1 and Mfn2 increase during the differentiation of SSCs, the differentiation of spermatogonia, and during meiosis I [ 22 ].…”

Section: Mfns In the Development Of Germ Cells And Embryosmentioning

confidence: 99%

Mitofusins: from mitochondria to fertility

Zhao

Heng

Wang

et al. 2022

Cell. Mol. Life Sci.

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Germ cell formation and embryonic development require ATP synthesized by mitochondria. The dynamic system of the mitochondria, and in particular, the fusion of mitochondria, are essential for the generation of energy. Mitofusin1 and mitofusin2, the homologues of Fuzzy onions in yeast and Drosophila, are critical regulators of mitochondrial fusion in mammalian cells. Since their discovery mitofusins (Mfns) have been the source of significant interest as key influencers of mitochondrial dynamics, including membrane fusion, mitochondrial distribution, and the interaction with other organelles. Emerging evidence has revealed significant insight into the role of Mfns in germ cell formation and embryonic development, as well as the high incidence of reproductive diseases such as asthenospermia, polycystic ovary syndrome, and gestational diabetes mellitus. Here, we describe the key mechanisms of Mfns in mitochondrial dynamics, focusing particularly on the role of Mfns in the regulation of mammalian fertility, including spermatogenesis, oocyte maturation, and embryonic development. We also highlight the role of Mfns in certain diseases associated with the reproductive system and their potential as therapeutic targets.

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“…We obtained the sequences for primer (F: 5'-CCT ACA AAC GCT CCT TCC ACT-3' , R: 5'-AGA GAA TAT AGG GCG GTG ATT ACT -3') and custom Taqman probe (FAM-TTG TTG GGG GTA GAG CTA AGT TGG T-MGBNFQ) sequences from reference [34].…”

Section: Real-time Quantitative Polymerase Chain Reactionmentioning

confidence: 99%

Tight gene co-expression in BCB positive cattle oocytes and their surrounding cumulus cells

Walker

Nix

Wilson

et al. 2022

Reprod Biol Endocrinol

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Background Cytoplasmic and nuclear maturation of oocytes, as well as interaction with the surrounding cumulus cells, are important features relevant to the acquisition of developmental competence. Methods Here, we utilized Brilliant cresyl blue (BCB) to distinguish cattle oocytes with low activity of the enzyme Glucose-6-Phosphate Dehydrogenase, and thus separated fully grown (BCB positive) oocytes from those in the growing phase (BCB negative). We then analyzed the developmental potential of these oocytes, mitochondrial DNA (mtDNA) copy number in single oocytes, and investigated the transcriptome of single oocytes and their surrounding cumulus cells of BCB positive versus BCB negative oocytes. Results The BCB positive oocytes were twice as likely to produce a blastocyst in vitro compared to BCB- oocytes (P < 0.01). We determined that BCB negative oocytes have 1.3-fold more mtDNA copies than BCB positive oocytes (P = 0.004). There was no differential transcript abundance of genes expressed in oocytes, however, 172 genes were identified in cumulus cells with differential transcript abundance (FDR < 0.05) based on the BCB staining of their oocyte. Co-expression analysis between oocytes and their surrounding cumulus cells revealed a subset of genes whose co-expression in BCB positive oocytes (n = 75) and their surrounding cumulus cells (n = 108) compose a unique profile of the cumulus-oocyte complex. Conclusions If oocytes transition from BCB negative to BCB positive, there is a greater likelihood of producing a blastocyst, and a reduction of mtDNA copies, but there is no systematic variation of transcript abundance. Cumulus cells present changes in transcript abundance, which reflects in a dynamic co-expression between the oocyte and cumulus cells.

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Concurrent Measurement of Mitochondrial DNA Copy Number and ATP Concentration in Single Bovine Oocytes

Cited by 6 publications

References 31 publications

Effect of maternal age on ATP content and distribution of mitochondria in bovine oocytes

Effect of maternal age on ATP content and distribution of mitochondria in bovine oocytes

Mitofusins: from mitochondria to fertility

Tight gene co-expression in BCB positive cattle oocytes and their surrounding cumulus cells

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