Three-Dimensional Micro-Computed Tomography of the Adult Mouse Ovary

Frontiers in Cell and Developmental Biology
Giulia FiorentinoMaurizio Zuccotti

Abstract

In the mouse ovary, folliculogenesis proceeds through eight main growth stages, from small primordial type 1 (T1) to fully grown antral T8 follicles. Most of our understanding of this process was obtained with approaches that disrupted the ovary three-dimensional (3D) integrity. Micro-Computed Tomography (microCT) allows the maintenance of the organ structure and a true in-silico 3D reconstruction, with cubic voxels and isotropic resolution, giving a precise spatial mapping of its functional units. Here, we developed a robust method that, by combining an optimized contrast procedure with microCT imaging of the tiny adult mouse ovary, allowed 3D mapping and counting of follicles, from pre-antral secondary T4 (53.2 ± 12.7 μm in diameter) to antral T8 (321.0 ± 21.3 μm) and corpora lutea, together with the major vasculature branches. Primordial and primary follicles (T1-T3) could not be observed. Our procedure highlighted, with unprecedent details, the main functional compartments of the growing follicle: granulosa, antrum, cumulus cells, zona pellucida, and oocyte with its nucleus. The results describe a homogeneous distribution of all follicle types between the ovary dorsal and ventral regions. Also, they show that each of the ei...Continue Reading

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Citations

Aug 13, 2021·Frontiers in Cell and Developmental Biology·Bikem Soygur, Diana J Laird

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Methods Mentioned

BETA
X-ray

Software Mentioned

LAS X Navigator
Avizo
Fiji ImageJ
RStudio
NRecon

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