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. 2021 Nov 4;9:749559. doi: 10.3389/fcell.2021.749559

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Copyright © 2021 Umer, Arévalo, Phadke, Lohanadan, Kirfel, Sons, Sofia, Witke and Schorle.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Impaired acrosome biogenesis in Pfn3-deficient mice. Adult testis sections of Pfn3^+/+ (left panel), Pfn3^+/– (middle panel), and Pfn3-deficient (right panel) mice; the developing acrosome was labeled with PNA-FITC (green) and cell nuclei were stained with DAPI (blue); inset panel showed the Golgi/cap/acrosomal phase of the tubules displayed in the main panel (n = 3). In the Golgi phase, proacrosomal granule (green) labeled by PNA-FITC for (A) Pfn3^+/+, (B) Pfn3^+/–, and (C) Pfn3^–/– round spermatozoa. In the cap phase, acrosomal caps (green) stained for (D) Pfn3^+/+, (E) Pfn3^+/–, and (F) Pfn3^–/– round spermatozoa (white arrows show fragmented cap structures). In the acrosome phase, PNA-FITC-labeled acrosomal area on (G) Pfn3^+/+, (H) Pfn3^+/–, and (I) Pfn3^–/– elongated spermatids. Scale bar = 20 μm. (J) Ultrastructure analysis using TEM revealed the acrosomal structures of Pfn3^+/+, Pfn3^+/–, and Pfn3^–/– sperm cells. Scale bar = 2 μm. (K) Immunofluorescence staining using PNA-FITC (green) on epididymal sperm cells of Pfn3^+/+, Pfn3^+/–, and Pfn3^–/– mice (n = 3). Scale bar = 20 μm. (L) TEM evaluation shows the percentage of malformed acrosomes in Pfn3^+/+, Pfn3^+/–, and Pfn3^–/– sperm cells (n = 2). (M) A graph represents the PNA-stained defective acrosome percentage of Pfn3^+/+, Pfn3^+/–, and Pfn3^–/– sperm cells (n = 3). Two hundred spermatozoa were counted per genotype.