Abstract
Simple SummaryStudies have revealed that the communication networks among epididymal epithelial cells play an essential role in sperm maturation and storage. Therefore, the localization and expression of V-ATPase and cytokeratin 5 in the clear cells and basal cells, respectively, of immature and mature goat epididymis was investigated. To the best of our knowledge, the present study is the first to use immunofluorescence labeling and confocal imaging to characterize the differentiation of clear cells and basal cells in the goat epididymis. The findings could help the understanding of the regulatory mechanism that creates an optimal luminal environment for sperm maturation and storage in the epididymis.The acidic luminal environment of the epididymis is regulated by the communication networks among epididymal epithelial cells; it is necessary for sperm maturation and storage. To characterize epididymal epithelial cell differentiation, the localization and expression of hydrogen-pumping vacuolar ATPase (V-ATPase) and cytokeratin 5 (KRT5) in the clear and basal cells, respectively, of immature and mature goat epididymis and vas deferens was examined. The epididymides and vas deferens were obtained from goats aged 1, 2, and 12–14 months. To assess the localization and expression patterns of V-ATPase and KRT5 in the caput, corpus, and cauda of the epididymis and proximal vas deferens, the tissue sections were subjected to immunofluorescence labeling and observed by confocal microscopy. Both clear and basal cells progressively started to differentiate in a retrograde manner. Clear cells disappeared from the cauda region after puberty, and they were maintained only in the caput and corpus regions of the adult goat epididymis. V-ATPase and KRT5 were co-expressed in the differentiated cells located at the base of the epithelium (i.e., basal cells). This cell type-specific differentiation and distribution of the epithelial cells plays a critical role in establishing a unique luminal environment for sperm maturation and storage in the goat epididymis.
Highlights
The epididymis is a highly convoluted duct that connects the testis and vas deferens (VD), and it is anatomically divided into the following three regions: the caput, corpus, and cauda
To confirm whether B1-VAPTase and KRT5 can be used as markers for clear cells (CCs) and basal cells (BCs), respectively, the epididymal goat tissues were double labeled with these markers
While B1-VATase was exclusively expressed in the cytoplasm of CCs (Figure 1A, white arrows) and at the base of the epithelium (Figure 1A, yellow arrowheads), and KRT5 was expressed in the cytoplasm of BCs located at the base of the epithelium (Figure 1B, white arrowheads)
Summary
The epididymis is a highly convoluted duct that connects the testis and vas deferens (VD), and it is anatomically divided into the following three regions: the caput, corpus, and cauda. Animals 2020, 10, 1273 and basal cells (BCs), which function cooperatively to establish an optimal luminal environment for the maturation, storage, protection, and transport of spermatozoa [1,2]. Principal cells are the most abundant cell type in the epididymis They secrete various proteins for sperm maturation and protection, and reabsorb and secrete bicarbonate and water via the cystic fibrosis transmembrane conductance regulator (CFTR) [3,4] and Aquaporin 9 (AQP9) [5,6], respectively, in the apical membrane of the epithelium. Basal cells, which are located at the base of the epithelium, are dome-shaped, but in rodents, BCs extend long cytoplasmic cell body projections into the lumen of the epididymis [12,13].
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