Foot, N.J.Gonzalez, M.B.Gembus, K.Fonseka, P.Sandow, J.J.Nguyen, T.T.Tran, N.Webb, A.I.Mathivanan, S.Robker, R.L.Kumar, S.2025-08-062025-08-062021Journal of Extracellular Vesicles, 2021; 10(8):e12113-1-e12113-212001-30782001-3078https://hdl.handle.net/2440/146566Extracellular vesicles (EVs) are important players in cell to cell communication in reproductive systems. Notably, EVs have been found and characterized in the male reproductive tract, however, direct functional evidence for their importance in mediating sperm function is lacking. We have previously demonstrated that Arrdc4, a member of the α-arrestin protein family, is involved in extracellular vesicle biogenesis and release. Here we show that Arrdc4-mediated extracellular vesicle biogenesis is required for proper sperm function. Sperm from Arrdc4–/– mice develop normally through the testis but fail to acquire adequatemotility and fertilization capabilities through the epididymis, as observed by reduced motility, premature acrosome reaction, reduction in zona pellucida binding and two-cell embryo production. We found a significant reduction in extracellular vesicle production by Arrdc4–/- epididymal epithelial cells, and further, supplementation of Arrdc4–/– sperm with additional vesicles dampened the acrosome reaction defect and restored zona pellucida binding. These results indicate that Arrdc4 is important for proper spermmaturation through the control of extracellular vesicle biogenesis.en© 2021 TheAuthors. Journal of Extracellular Vesicles published byWiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.arrestin; biogenesis; extracellular vesicles; fertility; sperm maturation; ubiquitin ligase adaptorsTestisEpididymisZona PellucidaSpermatozoaAcrosomeAnimalsMice, Inbred C57BLMice, Inbred CBAMiceIntracellular Signaling Peptides and ProteinsSperm MaturationSperm-Ovum InteractionsAcrosome ReactionFemaleMaleExtracellular VesiclesJournal article10.1002/jev2.12113579087Foot, N.J. [0000-0002-6567-9283]Tran, N. [0000-0002-4023-3373]Robker, R.L. [0000-0002-1538-4604]Kumar, S. [0000-0001-7126-9814]