Stereological study of organelle distribution in human oocytes at metaphase I

Coelho, Sofia; Luís, A.; Oliveira, Elsa; Alves, Ângela; Leal, Carla Bianca de Matos; Cunha, Mariana; Barreiro, Márcia; Barros, Alberto; Sá, Rosália; Sousa, Mário

doi:10.1017/s0967199420000131

Cited by 6 publications

(2 citation statements)

References 106 publications

(214 reference statements)

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“…The quantitative analysis of the 3D image data indicated that, in our samples of maturing human oocytes, mitochondrion occupies approximately 4.26% cytoplasm volume (Figure 2J). This result derived from densely recorded 3D datasets provide a more accurate estimate than the previous TEM stereological studies using 2D image data (Pires-Luís et al, 2016;Coelho et al, 2020).…”

Section: Resultsmentioning

confidence: 62%

High-Resolution 3D Reconstruction of Human Oocytes Using Focused Ion Beam Scanning Electron Microscopy

Trebichalská

Javůrek

Tatíčková

et al. 2021

Front. Cell Dev. Biol.

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The egg plays a pivotal role in the reproduction of our species. Nevertheless, its fundamental biology remains elusive. Transmission electron microscopy is traditionally used to inspect the ultrastructure of female gametes. However, two-dimensional micrographs contain only fragmentary information about the spatial organization of the complex oocyte cytoplasm. Here, we employed the Focused Ion Beam Scanning Electron Microscopy (FIB-SEM) to explore human oocyte intracellular morphology in three dimensions (3D). Volume reconstruction of generated image stacks provided an unprecedented view of ooplasmic architecture. Organelle distribution patterns observed in nine donor oocytes, representing three maturational stages, documented structural changes underlying the process by which the egg acquires developmental competence. 3D image segmentation was performed to extract information about distinct organelle populations, and the following quantitative analysis revealed that the mitochondrion occupies ∼ 4.26% of the maturing oocyte cytoplasm. In summary, this proof-of-concept study demonstrates the potential of large volume electron microscopy to study rare samples of delicate female gametes and paves the way for applying the FIB-SEM technique in human oocyte research.

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Section: Resultsmentioning

confidence: 62%

High-Resolution 3D Reconstruction of Human Oocytes Using Focused Ion Beam Scanning Electron Microscopy

Trebichalská

Javůrek

Tatíčková

et al. 2021

Front. Cell Dev. Biol.

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“…The quantitative analysis of the 3D image data indicated that, in our samples of MI and MII oocytes, mitochondrion occupies approximately 4.26 % cytoplasm volume (Figure 2J). This result provides a more accurate estimate than previous 2D stereological studies [7,8].…”

mentioning

confidence: 48%

High-resolution 3D reconstruction of human oocytes using FIB-SEM

Trebichalská

Javůrek

Tatíčková

et al. 2021

Preprint

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The oocyte plays a pivotal role in the reproduction of our species. Nevertheless, its biology remains poorly understood. Electron microscopy is traditionally used to inspect the ultrastructure of female gametes. However, two-dimensional micrographs contain only fragmentary information about the spatial organization of the complex oocyte cytoplasm. Here, we employed the Focused Ion Beam Scanning Electron Microscopy (FIB-SEM) to explore human oocyte intracellular morphology in three dimensions (3D). Volume reconstruction from high-resolution image stacks provided an unprecedented view of ooplasmic architecture. Organelle distribution patterns observed in 9 donor oocytes, representing 3 maturational stages, documented structural changes underlying the process by which the egg acquires developmental competence. 3D image segmentation was performed to extract information about distinct organelle populations. The quantitative analysis of the organelle abundance revealed that mitochondrion occupies ~ 4.26 % of the maturing oocyte cytoplasm. This proof-of-concept study demonstrates the potential of FIB-SEM imaging to study human oocyte morphology.

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All that glitters is not gold: a stereological study of human donor oocytes

Santos

Luís

Alves

et al. 2023

Zygote

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Summary Here we report a quantitative analysis of human metaphase II (MII) oocytes from a 22-year-old oocyte donor, retrieved after ovarian-controlled hyperstimulation. Five surplus donor oocytes were processed for transmission electron microscopy (TEM), and a stereological analysis was used to quantify the distribution of organelles, using the point-counting technique with an adequate stereological grid. Comparisons between means of the relative volumes (Vv) occupied by organelles in the three oocyte regions, cortex (C), subcortex (SC) and inner cytoplasm (IC), followed the Kruskal–Wallis test and Mann–Whitney U-test with Bonferroni correction. Life cell imaging and TEM analysis confirmed donor oocyte nuclear maturity. Results showed that the most abundant organelles were smooth endoplasmic reticulum (SER) elements (26.8%) and mitochondria (5.49%). Significant differences between oocyte regions were found for lysosomes (P = 0.003), cortical vesicles (P = 0.002) and large SER vesicles (P = 0.009). These results were quantitatively compared with previous results using prophase I (GV) and metaphase I (MI) immature oocytes. In donor MII oocytes there was a normal presence of cortical vesicles, SER tubules, SER small, medium and large vesicles, lysosomes and mitochondria. However, donor MII oocytes displayed signs of cytoplasmic immaturity, namely the presence of dictyosomes, present in GV oocytes and rare in MI oocytes, of SER very large vesicles, characteristic of GV oocytes, and the rarity of SER tubular aggregates. Results therefore indicate that the criterion of nuclear maturity used for donor oocyte selection does not always correspond to cytoplasmic maturity, which can partially explain implantation failures with the use of donor oocytes.

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Stereological study of organelle distribution in human oocytes at metaphase I

Cited by 6 publications

References 106 publications

High-Resolution 3D Reconstruction of Human Oocytes Using Focused Ion Beam Scanning Electron Microscopy

High-Resolution 3D Reconstruction of Human Oocytes Using Focused Ion Beam Scanning Electron Microscopy

High-resolution 3D reconstruction of human oocytes using FIB-SEM

All that glitters is not gold: a stereological study of human donor oocytes

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