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. 2019 Feb 4;9:1262. doi: 10.1038/s41598-018-37766-2

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Directionality of cumulus cell projections. (A) SEM image showing multiple cumulus cell bodies, zona pellucida, and part of the oocyte in cross-section. Five cells (colored) were chosen for reconstruction. Scale bar, 5 μm. Video 1 shows 202 serial sections of this area and highlights the green cell and all of its projections. (A’) High-magnification SEM image of the zona pellucida showing TZPs in cross-section. The colored TZPs originated from the cumulus cells chosen for reconstruction in A. Scale bar, 2 μm. (B) Reconstruction of 5 cumulus cell bodies from (A) and every cytoplasmic projection derived from them. Reconstruction is 28.4 × 24.6 × 18.2 μm (x, y, z), encompassing 405 serial sections (each, 45 nm-thick). A rotating view of this reconstruction can be seen in Video 2. (C,D) Front and side views of reconstructed cumulus cells found directly adjacent to the zona pellucida (C), or displaced by 1–2 cell diameters (D).