PFN4 is required for manchette development and acrosome biogenesis during mouse spermiogenesis
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Updated: Sep 1, 2025
A Seminiferous Tubule Squash Technique for the Cytological Analysis of Spermatogenesis Using the Mouse Model
Published on: February 6, 2018
Naila Umer1, Sharang Phadke1, Farhad Shakeri2,3
1Department of Developmental Pathology, Institute of Pathology, University Hospital Bonn, 53127 Bonn, Germany.
Abstract:
Profilin 4 (Pfn4) is expressed during spermiogenesis and localizes to the acrosome-acroplaxome-manchette complex. Here, we generated PFN4-deficient mice, with sperm displaying severe impairment in manchette formation. Interestingly, HOOK1 staining suggests that the perinuclear ring is established; however, ARL3 staining is disrupted, suggesting that lack of PFN4 does not interfere with the formation of the perinuclear ring and initial localization of HOOK1, but impedes microtubular organization of the manchette. Furthermore, amorphous head shape and flagellar defects were detected, resulting in reduced sperm motility. Disrupted cis- and trans-Golgi networks and aberrant production of proacrosomal vesicles caused impaired acrosome biogenesis. Proteomic analysis showed that the proteins ARF3, SPECC1L and FKBP1, which are involved in Golgi membrane trafficking and PI3K/AKT pathway, are more abundant in Pfn4-/- testes. Levels of PI3K, AKT and mTOR were elevated, whereas AMPK level was reduced, consistent with inhibition of autophagy. This seems to result in blockage of autophagic flux, which could explain the failure in acrosome formation. In vitro fertilization demonstrated that PFN4-deficient sperm is capable of fertilizing zona-free oocytes, suggesting a potential treatment for PFN4-related human infertility.
Insights
Profilin 4 (PFN4) deficiency severely impairs sperm manchette formation and acrosome biogenesis in mice. This leads to reduced motility and potential infertility, though zona-free oocyte fertilization remains possible.
Area of Science:
- Reproductive Biology
- Spermatogenesis
- Molecular Cell Biology
Background:
- Profilin 4 (PFN4) is crucial for sperm development, localizing to the acrosome-acroplaxome-manchette complex.
- Understanding PFN4's role is vital for addressing male infertility.
Purpose of the Study:
- To investigate the function of PFN4 in mouse spermatogenesis and sperm function.
- To elucidate the molecular mechanisms underlying PFN4 deficiency-induced infertility.
Main Methods:
- Generation of PFN4-deficient (Pfn4-/-) mice.
- Sperm morphology, motility, and acrosome biogenesis analysis.
- Immunofluorescence staining for key proteins (HOOK1, ARL3).
- Proteomic analysis of testicular tissue.
- In vitro fertilization assays.
Main Results:
- Pfn4-/- sperm exhibit defective manchette formation, disrupted microtubular organization, and abnormal head/flagellar morphology.
- Acrosome biogenesis is impaired due to disrupted Golgi networks and aberrant vesicle production.
- Elevated levels of proteins involved in Golgi trafficking and PI3K/AKT pathway, alongside inhibited autophagy, were observed.
- Pfn4-/- sperm can fertilize zona-free oocytes, indicating potential therapeutic applications.
Conclusions:
- PFN4 is essential for proper manchette formation, sperm head shaping, and acrosome biogenesis during spermiogenesis.
- PFN4 deficiency disrupts microtubular organization and Golgi function, leading to male infertility.
- Targeting PFN4 pathways may offer novel strategies for treating human infertility.
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