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Stereological study of organelle distribution in human oocytes at metaphase I

Published online by Cambridge University Press:  14 April 2020

Sofia Coelho
Affiliation:
Laboratory of Cell Biology, Department of Microscopy, Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto (UP), Rua Jorge Viterbo Ferreira, 228, 4050-313Porto, Portugal Department of Life Sciences, Faculty of Sciences and Technology, New Lisbon University (UNL), Campus da Caparica, 2829-516Lisbon, Portugal
Ana Sílvia Pires-Luís
Affiliation:
Laboratory of Histology and Embryology, Department of Microscopy, Institute of Biomedical Sciences Abel Salazar, University of Porto, Rua Jorge Viterbo Ferreira, 228, 4050-313Porto, Portugal Department of Pathology, Hospital Centre of Vila Nova de Gaia/Espinho, Unit 1, Rua Conceição Fernandes, 1079, 4434-502Vila Nova de Gaia, Portugal
Elsa Oliveira
Affiliation:
Laboratory of Cell Biology, Department of Microscopy, Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto (UP), Rua Jorge Viterbo Ferreira, 228, 4050-313Porto, Portugal Multidisciplinary Unit for Biomedical Research (UMIB), University of Porto, Portugal
Ângela Alves
Affiliation:
Laboratory of Cell Biology, Department of Microscopy, Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto (UP), Rua Jorge Viterbo Ferreira, 228, 4050-313Porto, Portugal Multidisciplinary Unit for Biomedical Research (UMIB), University of Porto, Portugal
Carla Leal
Affiliation:
Centre of Assisted Medical Procriation (CPMA), Maternal Child Centre of the North (CMIN), Hospital and University Center of Porto (CHUP), Largo da Maternidade de Júlio Dinis, 4050-651Porto, Portugal
Mariana Cunha
Affiliation:
Center for Reproductive Genetics A. Barros (CGR), Av. do Bessa, 240, 1° Dto. Frente, 4100–012Porto, Portugal
Márcia Barreiro
Affiliation:
Centre of Assisted Medical Procriation (CPMA), Maternal Child Centre of the North (CMIN), Hospital and University Center of Porto (CHUP), Largo da Maternidade de Júlio Dinis, 4050-651Porto, Portugal
Alberto Barros
Affiliation:
Center for Reproductive Genetics A. Barros (CGR), Av. do Bessa, 240, 1° Dto. Frente, 4100–012Porto, Portugal Department of Genetics, Faculty of Medicine, University of Porto (FMUP), Alameda Prof. Hernâni Monteiro, 4200–319Porto, Portugal Institute of Health Research and Innovation (IPATIMUP/i3S), University of Porto, Rua Alfredo Allen, 208, 4200–135Porto, Portugal
Rosália Sá
Affiliation:
Laboratory of Cell Biology, Department of Microscopy, Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto (UP), Rua Jorge Viterbo Ferreira, 228, 4050-313Porto, Portugal Multidisciplinary Unit for Biomedical Research (UMIB), University of Porto, Portugal
Mário Sousa*
Affiliation:
Laboratory of Cell Biology, Department of Microscopy, Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto (UP), Rua Jorge Viterbo Ferreira, 228, 4050-313Porto, Portugal Multidisciplinary Unit for Biomedical Research (UMIB), University of Porto, Portugal
*
Author for correspondence: Mário Sousa. Laboratory of Cell Biology, Department of Microscopy, Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto (UP), Rua Jorge Viterbo Ferreira, 228, 4050-313Porto, Portugal. Tel: +351 919974476. E-mail: [email protected]

Summary

We have previously presented a stereological analysis of organelle distribution in human prophase I oocytes. In the present study, using a similar stereological approach, we quantified the distribution of organelles in human metaphase I (MI) oocytes also retrieved after ovarian stimulation. Five MI oocytes were processed for transmission electron microscopy and a classical manual stereological technique based on point-counting with an adequate stereological grid was used. Kruskal–Wallis and Mann–Whitney U-tests with Bonferroni correction were used to compare the means of relative volumes (Vv) occupied by organelles. In all oocyte regions, the most abundant organelles were mitochondria and smooth endoplasmic reticulum (SER) elements. No significant differences were observed in Vv of mitochondria, dictyosomes, lysosomes, or SER small and medium vesicles, tubular aggregates and tubules. Significant differences were observed in other organelle distributions: cortical vesicles presented a higher Vv (P = 0.004) in the cortex than in the subcortex (0.96% vs 0.1%) or inner cytoplasm (0.96% vs 0.1%), vesicles with dense granular contents had a higher Vv (P = 0.005) in the cortex than in the subcortex (0.1% vs 0%), and SER large vesicles exhibited a higher Vv (P = 0.011) in the inner cytoplasm than in the subcortex (0.2% vs 0%). Future stereological analysis of metaphase II oocytes and a combined quantitative data of mature and immature oocytes, will enable a better understanding of oocyte organelle distribution during in vivo maturation. Combined with molecular approaches, this may help improve stimulation protocols and in vitro maturation methods.

Type
Research Article
Copyright
© Cambridge University Press 2020

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Footnotes

*

Both authors contributed equally to this work.

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