Abstract
Ankyrin proteins (ANKRD) are key mediators linking membrane and sub-membranous cytoskeletal proteins. Recent findings have highlighted a new role of ANKRD31 during spermatogenesis, elucidating its involvement in meiotic recombination and male germ cell progression. Following testicular differentiation, spermatozoa (SPZ) enter into the epididymis, where they undergo several biochemical and enzymatic changes. The epididymal epithelium is characterized by cell-to-cell junctions that are able to form the blood-epididymal barrier (BEB). This intricate epithelial structure provides the optimal microenvironment needed for epididymal sperm maturation. To date, no notions have been reported regarding a putative role of ANKRD31 in correct BEB formation. In our work, we generated an Ankrd31 knockout male mouse model (Ankrd31–/–) and characterized its reproductive phenotype. Ankrd31–/– mice were infertile and exhibited oligo-astheno-teratozoospermia (a low number of immotile SPZ with abnormal morphological features). In addition, a complete deregulation of BEB was found in Ankrd31–/–, due to cell-to-cell junction anomalies. In order to suggest that BEB deregulation may depend on Ankrd31 gene deletion, we showed the physical interaction among ANKRD31 and some epithelial junction proteins in wild-type (WT) epididymides. In conclusion, the current work shows a key role of ANKRD31 in the control of germ cell progression as well as sperm and epididymal integrity.
Highlights
Beyond the testis, spermatozoa (SPZ) travel into a long-convoluted tube, the epididymis, consisting of three main regions
In the current work, we reported the generation of an Ankrd31 knockout male mouse model (Ankrd31−/−) by using a clusters of regularly interspaced short palindromic repeats (CRISPR)/Cas9 genome editing strategy against exon 4 of the Ankrd31 gene
For lines 430 and 435, the sequences showed that the CRISPR/Cas9 cut exactly occurred as expected from the experimental design
Summary
Spermatozoa (SPZ) travel into a long-convoluted tube, the epididymis, consisting of three main regions (caput, corpus, and cauda). Luminal fluids, collected from the epididymis, show a high degree of compositional diversity, with a substantial segment-to-segment variation in inorganic ions, proteins, and small non-coding ribonucleic acid (RNA) transcripts that contribute to the regionalized functionality of this long tubule (Zhou et al, 2018). The BEB promotes epididymal lumen acidification, indispensable for: (i) sperm maturation, (ii) epididymosome transfer from epithelial cells to SPZ, and (iii) maintenance of sperm quiescence during epididymal transit (Sullivan, 2015; Zhou et al, 2018)
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