Immunohistochemical Studies of αSMA in the Epididymis of African Four-Toed Hedgehog (Atelerix albiventris)

The epididymis plays an important role in sperm maturation, storage, transport and in the secretion of enzymes and proteins into the tubular lumen. In this study, we examined the histology, microstereology and immunohistochemical localization of alpha smooth muscle (αSMA) in the three regions of the epididymis of the African four-toed hedgehog (Atelerix albiventris). Ten adult males were captured from the wild in Ibadan, Nigeria, between May and October, 2016. The animals were euthanized and the epididymis (caput, corpus and cauda regions) were retrieved and fixed in buffered neutral formalin ahead of the paraffin technique, following standard procedures. The duct of the epididymis was lined by pseudostratified columnar epithelium comprising basal, principal and apical cells as well as intraepithelial lymphocytes in proximity to basal cells. The principal cells, the major cells encountered within the epididymal epithelium of the animal, decreased in population from the caput to the cauda epididymidis while the apical cells were more abundant in the cauda epididymidis. Positive reactions to αSMA were observed in the peritubular muscular coat of the epididymal duct as well as blood vessels across the three regions of the epididymis with the caput and cauda epididymidis showing stronger positive reactions compared to the corpus epididymidis. This study demonstrated that the histology, microstereology as well as the cellular constituents of the epididymal duct of the Atelerix albiventris are similar to those of other mammals with a slight variation. It has also highlighted variation in the localization of αSMA across the regions of the epididymis of the animal.

Differential tissue-specific damage caused by bacterial epididymo-orchitis in the mouse

Ascending bacterial urinary tract infections can cause epididymo-orchitis. In the cauda epididymidis, this frequently leads to persistent tissue damage. Less coherent data is available concerning the functional consequences of epididymo-orchitis on testis and caput epididymidis. This in vivo study addresses the functional and spatial differences in responsiveness of murine epididymis and testis to infection with uropathogenic Escherichia coli (UPEC). Whole transcriptome analysis (WTA) was performed on testis, caput, corpus and cauda epididymidis of adult C57BL/6 J wildtype mice. Following UPEC-induced epididymo-orchitis in these mice, epididymal and testicular tissue damage was evaluated histologically and semi-quantitatively at 10 days and 31 days post-inoculation. Expression of inflammatory markers and candidate antimicrobial genes were analysed by RT-qPCR. WTA revealed distinct differences in gene signatures between caput and cauda epididymidis, particularly amonst immunity-related genes. Cellular and molecular signs of testicular inflammation and disruption of spermatogenesis were noticed at day 10, but recovery was observed by day 31. In contrast to the cauda, the caput epididymidis did not reveal any signs of gross morphological damage or presence of pro-inflammatory processes despite confirmed infection. In contrast to beta-defensins, known UPEC-associated antimicrobial peptides (AMP), like Lcn2, Camp and Lypd8, were inherently highly expressed or upregulated in the caput following infection, potentially allowing an early luminal protection from UPEC. At the time points investigated, the caput epididymidis was protected from any obvious infection/inflammation-derived tissue damage. Studies addressing earlier time-points will conclude whether in the caput epididymidis a pro-inflammatory response is indeed not essential for effective protection from UPEC.

Next-generation sequencing analysis reveals segmental patterns of microRNA expression in yak epididymis

MicroRNAs (miRNAs) have emerged as potent regulators of gene expression and are widely expressed in biological systems. In reproduction, they have been shown to have a significant role in the acquisition and maintenance of male fertility, whereby deletion of Dicer in mouse germ cells leads to infertility. Evidence indicates that this role of miRNAs extends from the testis into the epididymis, controlling gene expression and contributing to regional variations in gene expression. In this study, RNA sequencing technology was used to investigate miRNA expression patterns in the yak epididymis. Region-specific miRNA expression was found in the yak epididymis. In all, 683 differentially expressed known miRNAs were obtained; 190, 186 and 307 differentially expressed miRNAs were identified for caput versus corpus, corpus versus cauda and caput versus cauda region pairs respectively. Kyoto Encyclopedia of Genes and Genomes results showed endocytosis as the most enriched pathway across region pairs, followed by protein processing in the endoplasmic reticulum, phagosome, spliceosome and biosynthesis of amino acids in region pair-specific hierarchical order. Gene ontology results showed varied enrichment in terms including cell, biogenesis, localisation, binding and locomotion across region pairs. In addition, significantly higher miR-34c expression was seen in the yak caput epididymidis relative to the corpus and cauda epididymidis.

Evidence that glycosaminoglycan storage and collagen deposition in the cauda epididymidis does not impair sperm viability in the Mucopolysaccharidosis type I mouse model

Mucopolysaccharidosis type I (MPS I) is a lysosomal storage disease caused by a deficiency of the lysosomal hydrolase, α-L-iduronidase (IDUA). IDUA degrades heparan and dermatan sulfates, two types of glycosaminoglycan (GAG), important signalling and structural molecules of the extracellular matrix. Because many cell types store GAGs, MPS I has been investigated in human and animal models. Enzyme replacement therapy is available for MPS I patients and has improved their life expectancy, allowing them to achieve reproductive age. The aim of this study was to evaluate epididymal and sperm morphology and function in a murine model of MPS I. We used C57BL Idua+/+ and Idua−/− adult male mice (6 months old) to investigate epididymal morphology, sperm ultrastructure, GAG characterisation and mating competence. Epithelial GAG storage, especially in the cauda epididymidis, was seen in Idua−/− mice. Regardless of the morphologic change and GAG storage found in the cauda epididymis, sperm morphology and motility were normal, similar to wild types. In the interstitium, vacuolated cells were found in addition to deposits of GAGs. Mating was not impaired in Idua−/− males and litter sizes were similar between groups. At the time point of the disease evaluated, the deficiency in IDUA affected the morphology of the epididymis in male Idua−/− mice, whereas sperm appearance and motility and the male’s capacity to mate and impregnate females were preserved.

Seasonal expressions of luteinising hormone receptor, follicle-stimulating hormone receptor and prolactin receptor in the epididymis of the male wild ground squirrel (Spermophilus dauricus)

Luteinising hormone (LH), follicle-stimulating hormone (FSH) and prolactin (PRL) are pituitary-derived hormones and mediate their functions through LH receptor (LHR), FSH receptor (FSHR) and PRL receptor (PRLR) respectively. This study aimed to investigate the seasonal expression patterns of LHR, FSHR and PRLR in the epididymis of the male wild ground squirrel during the breeding and non-breeding seasons. Histologically, principal cells, basal cells, cilia and mature spermatozoa were found in the lumen of caput, corpus and cauda epididymidis in the breeding season, whereas in the non-breeding season, cilia and basal cells were rarely found and the epididymidal duct was devoid of spermatozoa. Immunohistochemical results showed that LHR, FSHR and PRLR were mainly present in the filamentous cytoplasm layer of epithelial cells of the caput, corpus and cauda epididymidis and FSHR and PRLR displayed stronger staining in the breeding season than in the non-breeding season. Furthermore, the mRNA and protein levels of FSHR and PRLR in all regions of epididymis as well as the levels of LHR in the caput and cauda epididymidis were higher during the breeding season. The protein levels of FSHR, LHR and PRLR were positively correlated with epididymal weight. Together, these results suggest that LHR, FSHR and PRLR may regulate epididymal functional changes in the male wild ground squirrel during its seasonal breeding cycle.

Modification of sperm fatty acid composition during epididymal maturation in bats

Biochemical properties of polyunsaturated fatty acids (PUFAs) are fundamental to sperm movements. Amongst all adjustments operated during epididymal maturation, sperm membrane lipid composition is remodelled. Specifically, the proportion of PUFAs usually increases from the caput towards the cauda epididymidis. In mammals, PUFAs are predominantly acquired through the diet, which can consequently impact male fertility. We aimed at analysing to what extent n-6 and n-3 PUFAs are incorporated into sperm in the Seba’s short-tailed bat (Carollia perspicillata), and at demonstrating the effect of the sperm fatty acid composition on sperm mobility. We therefore provided food varying in fatty acid composition to males of C. perspicillata and measured the fatty acid composition and mobility traits in spermatozoa collected from the caput and cauda epididymides. We found that n-6 and n-3 PUFAs and saturated fatty acids were significantly related to sperm velocity but not to the proportion of progressive sperm (i.e. motility). Concomitant to an increase in sperm velocity, the level of fatty acid saturation increased from the caput to the cauda epididymidis, while the proportion of PUFAs remained similar along the epididymis. A reduction in n-6 PUFAs counterbalanced an increase in n-3 PUFAs. The food treatments did not affect the sperm fatty acid composition. Our results suggest that a precise endogenous control rather than dietary effects determines sperm fatty acid composition in C. perspicillata.

Supplementation of cauda epididymal plasma improves sperm characteristics following liquid preservation of ram semen at 3–5°C

Mammalian spermatozoa remain immotile and metabolically inactive in the cauda epididymidis, thus maintaining fertility for several weeks. The aim of this study was to functionally characterise and evaluate the effect of cauda epididymal plasma (CEP) on liquid preservation of ram spermatozoa. Four experiments were conducted to investigate the effects of: (1) CEP and its fractions on sperm motility; (2) CEP (10%, 15%, 20% v/v) on liquid preservation of ram spermatozoa; (3) seminal plasma (SP; 20%, 30%, 50% v/v) on liquid-preserved spermatozoa; and (4) both CEP and post-storage SP treatment on sperm characteristics. Biochemical characterisation of ram CEP revealed high protein (30.9 mg mL−1), catalase (68.9 IU mL−1), alkaline phosphatase (17.5 IU mL−1) activities and total antioxidant capacity (1112 µM Trolox equivalent). Progressive motility of prewashed cauda spermatozoa was reduced (P < 0.05) by CEP or its protein-rich fraction compared with protein-free plasma or phosphate-buffered saline. After 48 h storage, total motility, rapid motility (average path velocity >75 µm s−1; 53.9%, 73.5% and 71.8% with 0, 15% and 20% CEP respectively) and straight line velocity (86.3, 102.1 and 102.4 µm s−1 with 0, 15% and 20% CEP respectively) were significantly (P < 0.05) higher in the CEP-treated groups than the control. Viability and acrosomal integrity were similar between groups; however, functional membrane integrity was higher (P < 0.05) in the 15% CEP-treated group. Treatment of liquid-preserved spermatozoa with either 20%, 30% or 50% SP improved (P < 0.05) rapid motility and kinematics at each time point of storage compared with control. In conclusion, liquid preservation of ram spermatozoa in the presence of 15% or 20% CEP and post-storage treatment with SP significantly improve sperm characteristics.

Rosmarinic acid reverses the effects of metronidazole-induced infertility in male albino rats

Rosmarinic acid (RA) is a natural antioxidant that has many biological activities. In the present study we investigated the potential of RA to reverse the negative effects of the widely used antibiotic and antiprotozoal agent metronidazole (MTZ), which is known to induce reversible male infertility. Two doses of RA (5 and 15 mg kg–1) were studied in sexually mature rats with and without MTZ-induced infertility. Rats were intraperitoneally injected with 5 mg kg–1 RA or 15 mg kg–1 RA (in distilled water) and, 45 min later, they were intraperitoneally injected with 40 mg kg–1 MTZ (in distilled water). Cauda epididymidal sperm suspensions were used to assess sperm count, motility and morphology. Histological and ultrastructural studies were performed on the testes and cauda epididymidis. In rats in which infertility was not induced, neither dose of RA affected the parameters assessed. However, in sexually mature rats in which infertility was induced by 40 mg kg–1 MTZ, RA at both 5 and 15 mg kg–1 ameliorated the damaging effects of MTZ on final bodyweight (30 days later), sperm motility and morphology. Only 5 mg kg–1 RA, and not 15 mg kg–1 RA, improved the harmful effects of MTZ on the sperm count and testis ultrastructure. The findings of the present study have considerable clinical implications and suggest a possible use for RA to reverse the negative effects of MTZ on male fertility, the male reproductive system and spermatogenesis.

Sleep restriction in Wistar rats impairs epididymal postnatal development and sperm motility in association with oxidative stress

Good sleep quality has a direct effect on the activity of the neuroendocrine–reproductive control axis and oxidative stress. Thus, the aim of the present study was to evaluate whether sleep restriction (SR) during the peripubertal period impaired the postnatal development of the epididymis in Wistar rats. After 21 days SR (18 h per day), epididymides were collected on Postnatal Day (PND) 62 for evaluation of oxidative stress markers, inflammatory profile, sperm count and histopathological and stereological analyses; in addition, the motility of spermatozoa from the vas deferens was examined. SR significantly increased lipid peroxidation and glutathione levels in the caput and cauda epididymidis, and increased levels of total radical-trapping antioxidant potential in the caput epididymidis only. Neutrophil migration to the caput or corpus epididymidis was decreased by SR, and the size of the luminal compartment in the 2A region and the epithelial compartment in the 5A/B region was also decreased. In these regions, there was an increase in the size of the interstitial compartment. The percentage of immotile spermatozoa was higher in the SR group. In conclusion, SR affects epididymal postnatal development, as well as sperm motility, in association with increased oxidative stress and a decrease in the size of the epithelial compartment in the cauda epididymidis.