Vitrification leads to transcriptomic modifications of mice ovaries that do not affect folliculogenesis progression |
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| Affiliation: | 1. Departamento de Ciências Naturais, Universidade Federal de São João del Rei. Praça Dom Helvécio, 74 - Dom Bosco, São João del-Rei, MG, 36301-160, Brazil;2. Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Pres. Antônio Carlos 6627, Belo Horizonte, MG, 31270-901, Brazil;3. Departamento de Zootecnia, ESALQ/Universidade de São Paulo, Av. Pádua Dias 11, Piracicaba, SP, 13418-900, Brazil;4. Embrapa Recursos Geneticos e Biotecnologia, PqEB, Av. W5 Norte, Brasília, DF, 70770-917, Brazil;1. Department of Urology, Krankenhaus Hietzing mit Neurologischem Zentrum Rosenhügel, Vienna, Austria;2. Department of Laboratory Medicine, Krankenhaus Hietzing mit Neurologischem Zentrum Rosenhügel, Vienna, Austria;3. Karl Landsteiner Institute for Clinical Risk Management, Vienna, Austria;4. Department of Urology, Royal Bournemouth and Christchurch Hospitals NHS Foundation Trust, Bournemouth, United Kingdom;5. Department of Surgery, University of Cambridge, Cambridge, United Kingdom;1. Department of Reproductive Center, First Affiliated Hospital ofGuangxi Medical University, Nanning, Guangxi, 530021, China;2. Department of Obstetrics and Gynecology, the Fourth Affiliated Hospital of Guangxi Medical University, Liuzhou, Guangxi, 545005, China;3. Reproductive Medicine Center, Liuzhou Maternal and Child Healthcare Hospital, Liuzhou, Guangxi, 545005, China;1. Faculty of Medicine, Universiti Teknologi MARA Sungai Buloh Campus, Selangor, Malaysia;2. I-PPerFORM, Universiti Teknologi MARA, Malaysia;1. Dep. Gyn. Endocrinology and Reproductive Medicine, University Hospital Giessen, Marburg, Germany;2. Institute of Medical Biometry and Epidemiology, Philipps University, Marburg, Germany;3. Clinic for Veterinary Obstetrics, Gynaecology, and Andrology, Chair for Molecular Reproductive Medicine, Justus-Liebig-University, Giessen, Germany;1. Department of Urology, St. Marianna University of Medicine, 2 -16-1 Sugao, Miyamae, Kawasaki, Kanagawa, 216-8511, Japan;2. College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-0890, Japan;3. Laboratory of Animal Reproduction, Faculty of Bioscience and Bioindustry, Tokushima University, Ishii, Myozai-gun, Tokushima, 779-3233, Japan;1. Department of Physiology. University of the Basque Country (UPV/EHU), Leioa, Spain;2. Department of Biochemistry and Molecular Biology. University of the Basque Country (UPV/EHU), Vitoria-Gasteiz, Spain;3. Department of Biochemistry and Molecular Biology. University of Southern Denmark. Odense. Denmark;4. Center for Reproductive Medicine and Infertility Quirón Bilbao, Bilbao, Spain |
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| Abstract: | Ovarian tissue cryopreservation is emerging as a promising alternative for fertility preservation of cancer survivors. To date, more than a hundred couples have successfully had babies using this procedure, although it is still considered experimental and demands further investigation. In this work, we evaluated the effects of vitrification, warming and autotransplantation procedures on the morphology and gene expression of murine ovaries. Ovaries were removed from adult female C57BL6 mice (n = 15), vitrified, warmed and autotransplanted (vitrified group), additionally, ovaries were autotransplanted without vitrification (control group, n = 15). After twenty days, grafted ovaries were harvested and used for histological and ultrastructural analysis, germinal vesicle (GV) oocyte collection, RNA sequencing, and Transmission Electron Microscopy (TEM). All classes of follicles and GV were observed in both control and vitrified/warmed transplanted ovaries, and the numbers of primordial, antral and atretic follicles were not different (p > 0.05). Using RNA-seq, we detected 16,602 vs 13,527 expressed genes in vitrified and control ovaries, respectively; and 623 significantly dysregulated genes (fold change >1.5; 332 up-regulated and 291 down-regulated). Cellular membranes, cytoskeletons, and extracellular matrices were found as the main functions of the differentially expressed genes. Moreover, vitrified samples also presented ultrastructural alterations in the cytoskeleton, cell junctions, and endoplasmic reticulum. Taken together, this work showed for the first time that ovarian cells might trigger a compensatory gene regulation mechanism to maintain cellular structure and folliculogenesis progression after vitrification and autotransplantation. |
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