Search

The formation of Wolffian ducts (WDs) includes three highly coordinated developmental stages, specification, elongation, and maintenance, prior to sexual differentiation. After their initial formation in both sexes, the WD epithelium and mesenchyme have sexually dimorphic fates depending on the presence of androgen action and local genetic programs. Postnatal differentiation of the epithelial lineage into principal, basal, and clear cells is controlled by androgens, lumicrine, and local factors. A lineage progression for epithelial cells is established. Aberrant cell differentiation is one underlying cause of WD abnormalities and associated pathologies in both men and women. Wolffian ducts (WDs) are the paired embryonic structures that give rise to internal male reproductive tract organs. WDs are initially formed in both sexes but have sex-specific fates during sexual differentiation. Understanding WD differentiation requires insights into the process of fate decisions of epithelial and mesenchymal cells, which are tightly coordinated by endocrine, paracrine, and autocrine signals. In this review, we discuss current advances in understanding the fate-decision process of WD epithelial and mesenchymal lineages from their initial formation at the embryonic stage to postnatal differentiation. Finally, we discuss aberrant cell differentiation in WD abnormalities and pathologies and identify opportunities for future investigations. [ABSTRACT FROM AUTHOR]

We evaluated the effects of feeding 6%, 12% or 18% crude glycerin, containing 80.5% glycerol, on testicular histomorphometry and markers of oxidative stress and on plasma testosterone concentrations in lambs. Body weight, testicular biometric measurements, gonadosomatic index and net weight of the testicles were higher for the treated groups (P <0.05) compared with a control group that did not receive dietary glycerin. The mean total length of seminiferous tubules was higher in the 6% group (P <0.05), while the mean total tubular and seminiferous epithelium volumes increased in all treated groups (P <0.05). The volume of Leydig cells increased in the 12% group, while their number per gram of testicle decreased (P <0.05). There was a decrease in mean nuclear diameter and mean volume of Leydig cells, and an increase in the mean number of these cells per gram of testicle, in the 18% group (P <0.05). Plasma testosterone concentrations were unaffected. There was desquamation of seminiferous epithelium and vacuolation of Sertoli cells in the treated groups. Variable degrees of spermatocyte necrosis and the presence of giant cells were seen in all groups and there was intense vacuolation of Sertoli cells in the 12% and 18% groups. Superoxide dismutase and catalase production increased most in the 12% and 18% groups (P <0.05), while glutathione production was higher in the 18% group (P <0.05). Mean nitric oxide concentration decreased in all treated groups (P <0.05), while malondialdehyde production was higher in the 18% group than in the control and 6% groups (P <0.05). We conclude that the inclusion of 6% glycerin in the diet of lambs results in changes in testicular morphology that have been previously associated with improved reproductive function, but without evidence of oxidative stress. [ABSTRACT FROM AUTHOR]

Polyunsaturated fatty acids (PUFAs) are essential for mammalian testis development and sperm function. However, PUFAs that are contained in linseed oil are easily oxidized in the diet and biohydrogenated in the rumen. In this study, we investigated the effect of linseed as a source of PUFAs on the antioxidant capacity and testis development in Hu lamb. Seventy-five 3-month-old lambs were randomly assigned to three groups. Within each treatment group, 25 lambs were allocated to five pens (five lambs per pen). The lambs in the control group were fed a control diet without linseed for 42 days from D22 to D63. Group I (BS28) was fed a control diet from D22 to D35 and 8% linseed diet from D36 to D63. Group II (BS42) was fed an 8% linseed diet for 42 days from D22 to D63. After 63-day feeding trial, all lambs except the heaviest and lightest in each pen were humanely slaughtered and investigated. Results revealed that feeding linseed did not affect the body weight, scrotal circumference, and testis weight, whereas feeding linseed for 42 days increased the epididymis weight (37.85 ± 1.61 g vs. 32.09 ± 1.06 g, P < 0.05) compared with the control group. Feeding lambs with linseed for 42 days also significantly upregulated the expression of antioxidative (glutathione peroxidase 4 and copper-zinc superoxide dismutase), steroidogenesis (3β-hydroxysteroid dehydrogenase and steroid acute regulatory protein), and PUFA metabolism-related genes (fatty acid desaturase 2 and elongation of very long-chain fatty acid protein 2) and proliferating cell nuclear antigen mRNA (P < 0.05). It also increased the relative expression of mitochondrial DNA (P < 0.05), total antioxidant capacity (0.230 ± 0.019 mmol/mgprot vs. 0.175 ± 0.011 mmol/mgprot, P < 0.05), and superoxide dismutase (1661.467 ± 147.117 U/mgprot vs. 1158.891 ± 98.850 U/mgprot, P < 0.05) in testicular tissue but decreased the cholesterol concentration (0.331 ± 0.073 mmol/mgprot vs. 0.671 ± 0.092 mmol/mgprot, P < 0.05) compared with the control group. Therefore, feeding lambs with linseed for 42 days stimulated seminiferous tubule development and increased the number of Sertoli cells (20.71 ± 0.89 vs. 17.6 ± 0.73, P < 0.05), epididymal cauda lumina diameter (638.26 ± 22.32 μm vs. 444.41 ± 34.80 μm, P < 0.05), and the number of sperm in the epididymal cauda (68.91 ± 7.06 × 108/g vs. 36.61 ± 7.50 × 108/g). All these results suggested that feeding linseed in the early reproductive development stage of lambs upregulated the expression of antioxidative, steroidogenesis, and PUFA metabolism-related genes; increased the antioxidant capacity in lamb's testis; and contributed to testis development and spermatogenesis. Image 1 • Feeding linseed for 42 days increasing antioxidant capacity in testis of Chinese Hu lamb. • Feeding linseed up-regulated GPX4, Cu-ZnSOD, 3βHSD, StAR, FADS2, ELOVL2 and PCNA mRNA expression. • Feeding linseed stimulated seminiferous tubule development and increased the number of sperm in epididymal cauda. [ABSTRACT FROM AUTHOR]

Abstract Hippo signaling pathway is essential for tissue development and homeostasis, while its specific role in male reproductive tract development is still unclear. The objective of this study is to elucidate the localization and expressions of key Hippo pathway components (MST1/2, LATS1/2 and YAP1) in male reproductive tract (testis, epididymis, and ductus deferens) of prepubertal (3-month-old) and postpubertal (9-month-old) Hu sheep, as well as in the cauda epididymal and ejaculated spermatozoa. Results showed that the Hippo pathway proteins were diversely localized in male reproductive tract portions, and most of their expression levels increased during sheep testicular maturation. In addition, these Hippo components were mainly localized and highly expressed in ejaculated spermatozoa compared with cauda epididymal spermatozoa. In ejaculated spermatozoa, LATS1 was localized in the acrosomal head region, and LATS2 and YAP1 were expressed in the midpiece part. Taken together, the presence of Hippo signaling cascade in the pubertal development of male reproductive tract and spermatogenesis of Hu sheep, provides new insights on the function of these components in the process of male sexual maturation, capacitation and fertilization. Highlights • Hippo pathway proteins were diversely localized in male reproductive tract. • Expressions of Hippo pathway components (except for MST1) were elevated with sexual maturity. • Expressions of Hippo pathway components were correlated to spermatogenesis and sperm function. [ABSTRACT FROM AUTHOR]

Sunflower seed husk (SH) and rape straw (RS) are widely used in mutton sheep husbandry in the Hexi Corridor of China. This study explored whether or not a pre-puberty lamb diet consisting of sunflower SH and RS affects ram testis development. Fifty 60-day-old healthy Hu ram lambs (average body weight of 22.3 ± 0.1 kg) were randomly divided into two groups. In each treatment group, 25 lambs were allocated to five pens (five lambs per pen). The SH group received a diet containing 20 % SH, and the RS group received a diet containing 20 % RS. After a 77-day feeding trial, three lambs, except for the heaviest and lightest in each pen, were humanely slaughtered and investigated. Results revealed that the two groups had no difference in body weight (39.23 ± 0.52 kg vs. 38.53 ± 0.46 kg, P = 0.321). However, the testis weight (288.74 ± 23.45 g vs. 215.18 ± 17.96 g), volume (294.29 ± 25.39 mL vs. 208.67 ± 16.63 mL), epididymis weight (38.38 ± 1.44 g vs. 33.86 ± 1.37 g), testicular index (0.74 ± 0.06 vs 0.56 ± 0.05), and epididymis sperm count (4.88 ± 0.92 ×109/g vs. 2.27 ± 0.69 ×109/g) in the SH group were significantly higher than those in the RS group (P < 0.05). SH reduced the mtDNA relative copy number (P < 0.01) in the testis and increased the total antioxidant capacity (3.41 ± 0.94 U/mg prot vs. 1.54 ± 0.38 U/mg prot, P = 0.05). RNA sequencing results showed 50 differentially expressed genes, with 27 genes upregulated and 23 genes downregulated in the SH group compared with the RS group. KEGG analysis revealed that the pathways related to polyunsaturated fatty acid metabolism, and vitamin digestion and absorption were enriched. The findings suggest that feeding SH in the early reproductive development stage of lambs is more beneficial for testis development and spermatogenesis compared with feeding RS. • Feeding sunflower seed husk during the peri-puberty was more conducive to ram reproductive than feeding rape straw. • Feeding sunflower seed husk stimulated testis development and increased the number of sperm in the epididymis. • The pathways related to polyunsaturated fatty acid metabolism, vitamin digestion and absorption were enriched. [ABSTRACT FROM AUTHOR]

The tissue-specific profile of alternatively spliced genes (ASGs) and their involvement in reproduction processes characteristic of turkey testis, epididymis, and ductus deferens were investigated for the first time in birds. Deep sequencing of male turkey reproductive tissue RNA samples (n = 6) was performed using Illumina RNA-Seq with 2 independent methods, rMATs and SUPPA2, for differential alternative splicing (DAS) event prediction. The expression of selected ASGs was validated using quantitative real-time reverse transcriptase-polymerase chain reaction. The testis was found to be the site of the highest number of posttranscriptional splicing events within the reproductive tract, and skipping exons were the most frequently occurring class of alternative splicing (AS) among the reproductive tract. Statistical analysis revealed 86, 229, and 6 DAS events in the testis/epididymis, testis/ductus deferens, and epididymis/ductus deferens comparison, respectively. Alternative splicing was found to be a mechanism of gene expression regulation within the turkey reproduction tract. In testis, modification was observed for spermatogenesis specific genes; the changes in 5' UTR could act as regulator of MEIG1 expression (a player during spermatocytes meiosis), and modification of 3' UTR led to diversification of CREM mRNA (modulator of gene expression related to the structuring of mature spermatozoa). Sperm tail formation can be regulated by changes in the 5' UTR of testicular SLC9A3R1 and gene silencing by producing dysfunctional variants of ODF2 in the testis and ATP1B3 in the epididymis. Predicted differentially ASGs in the turkey reproductive tract seem to be involved in the regulation of spermatogenesis, including acrosome formation and sperm tail formation and binding of sperm to the zona pellucida. Several ASGs were classified as cilia by actin and microtubule cytoskeleton organization. Such genes may play a role in the organization of sperm flagellum and post-testicular motility development. To our knowledge, this is the first functional investigation of alternatively spliced genes associated with tissue-specific processes in the turkey reproductive tract. [ABSTRACT FROM AUTHOR]

Osteopontin (OPN) is indispensable in mammalian reproduction, but the role of OPN in male reproductive tract and fertility remains unclear. The objective of this study is to elucidate the function of OPN by unveiling the localization and expression of OPN in the reproductive tract (testis, epididymis, and ductus deferens) of male Hu sheep in different ages (10-days, 4-months, and 8-months). To accomplish this, the localization, mRNA and protein expression patterns of OPN in all samples were investigated. Immune staining showed that OPN was present in the testicular interstitium of prepubertal Hu sheep testis (10-days and 4-months group), while it was immunostained in acrosomes of spermatids nearby adluminal compartment of seminiferous tubules in sexual maturity Hu sheep testis (8-months group). The localization of OPN in epididymis gradually changed from the loose connective tissue to the apical region of principal cells (pseudostratified columnar epithelium) with growing (10-days to 8-months). In addition, increase trend was observed in the mRNA expression levels of OPN with growing in the same reproductive tissues ( P < 0.05). Furthermore, two different OPN isoforms of 30 kDa and 34 kDa were detected in the reproductive tract of male Hu sheep by western blot. Immunofluorescence detection showed that OPN was localized in the cauda epididymal spermatozoa. These results suggested that the expression of OPN might be closely related to spermatogenesis and spermatozoa function in Hu sheep. This will be helpful for us to understand how OPN regulate the high reproductive capacity in Hu sheep. [ABSTRACT FROM AUTHOR]

External and internal penile morphologies have evolved rapidly and divergently in many mammalian orders and are extremely useful for taxonomic studies, particularly in the recovery of phylogenetic relationships. Eptesicus furinalis , a Vespertilionid bat, belongs to a taxon in which species recognition can be difficult when only traditional features are employed. Therefore, any feature that may contribute to the more accurate characterization of this taxon is relevant. In this study, we describe the histomorphology of the penis and baculum of this species after analyzing serial transverse sections and three-dimensional (3D) reconstructions. The glans penis was small with no epidermal projections and had an inverted Y-shaped baculum for most of its length. Internally, the penis contained three erectile tissues: corpus cavernosum, accessory cavernous tissue, and corpus spongiosum around the urethra. The internal anatomy of the E. furinalis penis displayed the basic vascular mammalian pattern but had certain features that were unique to this species, such as accessory cavernous tissue located dorsal and lateral to the tunica albuginea of the corpora cavernosa, a glans penis with a cone morphology, and a marked development of spongy tissue. [ABSTRACT FROM AUTHOR]

Abstract: The aim of this study was to investigate the effects of caffeine (20mg/L) intake on cadmium (15mg/L) accumulation in the rat blood, testes, epididymis and prostate as well as cadmium-induced changes to the antioxidant defense system of the epididymis. Caffeine reduced the cadmium concentration in all tissues analyzed. Meanwhile, cadmium reduced catalase activity and increased superoxide dismutase (SOD) activity in the epididymis. Caffeine increased SOD activity, catalase and glutathione tissue expression and sustains the cadmium''s effect on catalase and GSP-Px activity. No differences in the expression of metallothionein and lipid peroxidation were observed among the different treatments in the epididymis. In conclusion, low doses of cadmium alter the antioxidant enzymatic profile of the epididymis, but not induced oxidative lipid damage. Caffeine intake reduces overall cadmium accumulation in the organism and enhances the levels of antioxidant protein expression in the epididymis, thus exerting a protective effect against this metal. [Copyright &y& Elsevier]

Abstract: The objective of this study was to examine whether domestic fowl (Gallus domesticus) sperm undergo maturation in their capacity for survival and fertilization in the male reproductive tract. Sperm collected from the testis, epididymis and the proximal, middle and distal vas deferens were simultaneously stored in vitro in minimum essential medium (MEM) at 39°C for 0, 3 and 6h, and at 4°C for 24 and 48h. Sperm membrane integrity was measured using the dual fluorescent stain SYBR-14/propidium iodide (PI). Aliquots of sperm from the various sites were subjected to artificial insemination (AI) into the uteri of hens to assess the duration of sperm survival in the oviduct and to determine the fertility status of the sperm. Testicular sperm exhibited a very low capacity to survive under in vitro liquid storage conditions, irrespective of the storage temperature used, and in the oviduct, and they had a low ability to fertilize the ovum. On the contrary, sperm from the distal vas deferens had a higher survival rate during in vitro storage periods, a longer life span in the oviduct, and high fertility. Survival and fertilizing capacity of the sperm recovered from the testes increased gradually (P <0.05) from the testes to the distal vas deferens. In conclusion, we suggest that fowl sperm may undergo functional maturation through a process of gradual changes in their survival and fertilization capacities during their passage through the successive parts of the male reproductive tract. [Copyright &y& Elsevier]

Background: It is debated whether chronic urogenital inflammations and infections may trigger the formation of antisperm antibodies (ASA) in semen. Objective: To evaluate the formation of ASA in defined chronic inflammatory and infectious diseases of the male reproductive tract (MRT). Design, setting, and participants: Three hundred sixty-five patients retrospectively enrolled in a single center were categorized as having National Institutes of Health (NIH) category II chronic prostatitis (n =38), NIH category IIIa chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) (n =59), NIH category IIIb CP/CPPS (n =213), chronic epididymitis (n =34), and chronic urethritis (n =21). Forty-five age-matched men served as controls. Measurements: All subjects underwent microbiologic and cytologic analysis for common bacteria, yeasts, and mycoplasma using the four-glass test. Urine samples, ejaculates, and urethral swabs were analyzed with polymerase chain reaction (PCR) for Chlamydia trachomatis and Neisseria gonorrhea. Semen analysis followed World Health Organization (WHO) standards. ASA in seminal plasma were analyzed using the mixed agglutination reaction (MAR) test. Results and limitations: The overall positive detection rate of clinically significant levels (≥50% of spermatozoa coated by ASA) of IgG and IgA antibodies was 1.8% and 0.8%, respectively, in the patient group. No clinically significant levels of ASA were detected in the control group, and no statistically significant difference was observed between controls and patients (IgG, p =1.0; IgA, p =1.0). No difference was found between the different inflammatory and infectious diseases and the control group in the detection rate of ASA, even when the cut-point value was lowered to ≥1% (IgG, p =0.4; IgA, p =0.3). Moreover, in one selected subgroup of patients (n =26) with persistent increased inflammatory parameters (peroxidase-positive leukocytes [PPL] ≥1×106/ml and elastase ≥230ng/ml), no significant difference in the levels of ASA was observed compared with the controls (IgG, p =0.1; IgA, p =0.8). Conclusion: There is no association between chronic inflammatory or infectious diseases of the MRT and the presence of ASA in semen. [Copyright &y& Elsevier]

ABSTRACT The aim of this study was to determine whether chicken sperm express avian 13-defensin 3 (AvBD-3), an antimicrobial peptide, and retain it dur- ing the residence in the male and female reproductive tract. Immunocytochemistry to identify AvBD-3 was performed in the testis, epididymis, ductus deferens, ejaculated sperm, and utero-vaginal junction of the ovi- duct of inseminated hens. Reverse transcription-PCR analysis was also performed to confirm the synthesis of AvBD-3 by testicular cells and ejaculated sperm. The immunoreaction products for AvBD-3 were identified in the elongated spermatids attached to the seminiferous epithelium and in the sperm in the lumen of seminifer- ous tubules. Immunoreaction products were found in the sperm in the epididymal ducts and ductus defer- ens. In the ejaculated sperm, immunoreaction products were distributed in the midpiece and cephalic region of the tail of sperm, but not in the head. The AvBD- 3 immunoreaction products were also identified in the sperm stored in the sperm storage tubules in the utero- vaginal junction. Reverse transcription-PCR analysis showed the expression of AvBD-3 mRNA in the testicu- lar tissue, but not in the ejaculated sperm. These re- sults suggest that AvBD-3 is synthesized by late stage of spermatids in the testis, and this molecule is retained by the sperm during the passage of male reproductive tract, and even in the sperm storage tubules of oviduct. The sperm AvBD-3 is assumed to play roles in protect- ing the sperm from the microbial infection and enable to survive in the male and female reproductive tracts. [ABSTRACT FROM AUTHOR]

Abstract: Male reproductive tract CD52 (mrt-CD52) is known to be a pathogenic antigen for immunological infertility. Although human CD52 has been extensively investigated, the properties of mouse CD52 are not well elucidated. This study was conducted, therefore, to examine the tissue distribution, molecular composition and immunogenicity of mouse mrt-CD52. Immunohistological studies with an antibody to a synthetic peptide showed that mouse CD52 was localized mainly in the cauda epididymis and vas deferens, but not in the testis, liver, kidney or spleen. The molecule was composed of Asn (N)-linked and The/Ser (O)-linked carbohydrates as well as a glycosylphosphatidyl (GPI) anchor portion. Purified mrt-CD52 preparations produced antibodies by subcutaneous and intranasal immunization in both male and female mice. These antisera showed sperm-immobilizing activities with complement to mouse sperm. The research indicated mouse CD52 had similar biochemical and immunological properties to human CD52. This animal experiment is a good model for investigating human mrt-CD52 antibody detected in infertile patients. [Copyright &y& Elsevier]

Abstract: Increases in human male reproductive disorders (testicular cancer, cryptorchidism, hypospadias, and low sperm counts) might stem from increased exposure of the developing male to environmental estrogens. In the present study, we investigated the effects of octylphenol (OP), an estrogenic compound, exposure on the male reproductive system during the fetal period in which the development of reproductive organs and sexual differentiation occurs. Male rats were treated with OP in utero at doses of control (vehicle), 100 or 250mg/kg/day. After birth, male rats were allowed to grow until adulthood, and then testes, epididymides, and prostate glands were investigated histopathologically. Sperm counts and percentage of abnormal sperm were determined. Seminiferous and epididymal round tubules were evaluated for tubule diameter, lumen diameter, and height of tubule epithelium. Treatment with OP caused abnormalities in the histology of the testis and epididymis and induced atrophy of prostate glands tubules. Although there were no differences in sperm counts among treatment groups, abnormal sperm percentages in the high dose group increased significantly. The results of this study demonstrate that maternally injected OP causes adverse effects on male reproductive tract at adulthood, especially on sperm structure. [Copyright &y& Elsevier]

Abstract: The aim of the present study was to identify the muscarinic acetylcholine receptor (mAChR) mRNA subtypes in the rat seminal vesicle. Furthermore, the mAChR subtypes involved in the contraction of the seminal vesicle were also explored. Reverse transcriptase-polymerase chain reaction (PCR) was performed and five PCR products corresponding to M1–M5 mAChR mRNA subtypes were detected in this tissue. Functional pharmacological studies indicated that the rank order of mAChR antagonists in blocking the contractile effects of carbachol was p-fluoro-hexahydro-sila-difenidol (pF-HHSiD)≫tropicamide>methoctramine=pirenzepine. This antagonist profile indicates that M3 mAChR subtype is predominantly involved in the seminal vesicle contraction. Furthermore, immunohistochemical studies confirmed the presence of the M3 mAChR subtype in the smooth muscle layers. M2 mAChR subtype was also immunolocalized in smooth muscle cells and may be involved in the contraction of this tissue. The presence of M2 and M3 mAChR subtypes in the epithelial cells suggests that these receptors could be involved in the protein secretion. Taken together, the cholinergic neurotransmitter may be a factor controlling contractility and protein secretion in this tissue. [Copyright &y& Elsevier]

Abstract: Male fertility is a highly controlled process that allows proliferation, meiosis and differentiation of male germ cells in the testis, final maturation in the epididymis and also requires functional male accessory glands: seminal vesicles, prostate and corpus cavernosum. In addition to classical endocrine and paracrine controls, mainly by gonadotropins LH and FSH and steroids, there is now strong evidence that all these processes are dependent upon the presence of homocellular or heterocellular junctions, including gap junctions and their specific connexins (Cxs), between the different cell types that structure the male reproductive tract. The present review is focused on the identification of Cxs, their distribution in the testis and in different structures of the male genital tract (epididymis, seminal vesicle, prostate, corpus cavernosum), their crucial role in the control of spermatogenesis and their implication in the function of the male accessory glands, including functional smooth muscle tone. Their potential dysfunctions in some testis (spermatogenic arrest, seminoma) and prostate (benign hyperplasia, adenocarcinoma) diseases and in the physiopathology of the human erectile function are also discussed. [Copyright &y& Elsevier]

Abstract: The immediate and delayed effects of prepubertal exposure to di(2-ethylhexyl)phthalate (DEHP) or oestradiol benzoate on the plasma concentrations of testosterone, oestradiol and LH, as well as testicular morphology were examined in prepubertal boars. In a split litter design experiment, prepubertal boars were intramuscularly exposed to DEHP, oestradiol or vehicle during five weeks, starting at six weeks of age. The dose of DEHP was 50mg/kg of bodyweight twice weekly, which is in the same range as recently used oral doses in rodents. Oestradiol-benzoate was administered at 0.25mg/kg of bodyweight twice weekly. One set of animals was examined immediately after the exposure, and the other set was examined at an age of 7.5 months. During the exposure period concentrations of LH in plasma were lower (p =0.02) in the oestradiol-treated animals than in the control group. In the group exposed to oestradiol, the relative to the body weight of the testicles tended to be lower (p =0.07) than control immediately after five weeks of exposure, and the relative to the body weight of the seminal vesicles tended to be lower (p =0.05) than control at 7.5 months of age. In the DEHP-exposed group an elevated (p =0.005) concentration of testosterone and increased (p =0.04) area of the Leydig cells in the testicles compared to the control group were seen at 7.5 months of age. These data suggest that DEHP early in life causes delayed effects on the reproductive system in the adult. [Copyright &y& Elsevier]

Abstract: This study examined proteolytic enzymes and serine proteinase inhibitors in turkey seminal plasma with relation to their distribution within the reproductive tract and to yellow semen syndrome (YSS). Proteases of blood plasma, extracts from the reproductive tract, and seminal plasma were analyzed by gelatin zymography. We found a clear regional distribution of proteolytic enzymes in the turkey reproductive tract. Each part was characterized by a unique profile of serine proteolytic enzymes of molecular weights ranging from 29 to 88kDa. The ductus deferens was found to be a site of very intense proteolytic activity. Two metalloproteases of 58 and 66kDa were detected in all parts of the reproductive tract and seminal plasma. Using electrophoretic methods for detection of anti-trypsin activity, we found three serine proteinase inhibitors in turkey seminal plasma. Two inhibitors were found in the testis and epididymis and a third in the ductus deferens and seminal plasma. Blood plasma was characterized by the presence of two metalloproteinases and one serine proteinase inhibitor (of low migration rate) that were also detected in the reproductive tract. Amidase and anti-trypsin activities (expressed per gram of protein) differed for yellow and white seminal plasma. We concluded that turkey seminal plasma contains metalloproteases, serine proteinases, and serine proteinase inhibitors. The metalloproteases and one proteinase inhibitor are related to blood proteinases but the other two inhibitors and serine proteinases seem to be unique for the reproductive tract. [Copyright &y& Elsevier]

We previously identified two murine secretory proteins, mE-RABP(Lcn5) and mEP17(Lcn8), belonging to the lipocalin family and specifically expressed in the epididymis. The genes are contiguous and localized on mouse chromosome 2. We now show that five other related lipocalin genes, Lcn9, Lcn10, Lcn11, Lcn12, and Lcn13, that evolved by in situ tandem duplication are present on the same locus. Lcn9, Lcn10, Lcn12, and Lcn13 genes, like Lcn5 and Lcn8 genes, are specifically expressed in the mouse epididymis. However, each gene has a distinct spatial expression within the epididymis and different regulation. Analysis of the human genome sequence shows the presence of genes encoding lipocalins with genomic organization, chromosomal arrangement, and orientation similar to that of the corresponding murine genes, indicating that the epididymal cluster is evolutionary conserved. A phylogenetic analysis of the new epididymal proteins reveals their spread position in the lipocalin protein family tree. This suggests the preservation of the regulatory sequences, while protein sequences have greatly diverged, reflecting functional diversity and possibly multifunctionality. In terms of the cluster ancestry, epididymal expression possibly appeared in a PGDS-like lipocalin in amniotes, and the duplications generating the cluster occurred at least in the common ancestor of rodents and primates. The presence and conservation of a cluster of five genes encoding epididymal lipocalins, differently regulated and regionalized in the epididymis, strongly suggests that these proteins may play an important role for male fertility. [Copyright &y& Elsevier]

Adult male rats previously exposed on gestation days (GD) 12–21 to di(n-butyl) phthalate (DBP) have reproductive tract malformations, particularly agenesis of the epididymis, decreased sperm production, and Leydig cell hyperplasia and adenomas. Although similar effects are produced by the potent androgen receptor (AR) antagonist flutamide and are indicative of disruption of male sexual differentiation via an antiandrogenic mechanism, DBP is not an AR antagonist. The purpose of the study was to determine whether DBP causes pathologic changes and alterations in androgen status in the testis during the prenatal period of male reproductive tract differentiation. Pregnant CD rats were given corn oil, DBP (500 mg/kg/day), or flutamide (100 mg/kg/day) p.o. on GD 12–21. At GD 16–21, DBP caused hyperplasia of Leydig cells, many of which were 3β-hydroxysteroid dehydrogenase- and/or AR-positive. Focal areas of hyperplasia had increased numbers of Leydig cells positive for proliferating cell nuclear antigen (PCNA). At GD 21, testis atrophy was apparent, seminiferous cords in DBP-exposed fetuses were enlarged and contained multinucleated gonocytes that, unlike controls, were PCNA-positive. DBP, but not flutamide, markedly decreased testicular testosterone levels at GD 18 and 21. Fewer epididymal ducts and reduced AR staining in some ducts were evident with DBP treatment, whereas decreased overall AR staining was seen with flutamide in the presence of mild Leydig cell hyperplasia. Leydig cell proliferation is likely a compensatory mechanism to increase testicular steroidogenesis triggered by testosterone insufficiency. The overall decrease in androgen concentration is not corrected and results in reproductive tract malformations. The multinuclearity and proliferation of gonocytes suggests an underlying Sertoli cell dysfunction. [Copyright &y& Elsevier]

• Multiple molecular initiating events lead to adverse male reproductive development. • Rat dams exposed to mixture of 9 phthalates, 5 pesticides, and 1 pesticide metabolite. • Dose structure based on individual chemical NOAELs specific for male reproductive effects. • Mixture produced numerous male reproductive adverse outcomes below chemical NOAELs. • Cumulative assessment grouping should consider chemicals with common adverse outcomes. Humans carry residues of multiple synthetic chemicals at any given point in time. Research has demonstrated that compounds with varying molecular initiating events (MIE) that disrupt common key events can act in concert to produce cumulative adverse effects. Congenital defects of the male reproductive tract are some of the most frequently diagnosed malformations in humans and chemical exposures in utero can produce these effects in laboratory animals and humans. Here, we hypothesized that in utero exposure to a mixture of pesticides and phthalates, each of which produce male reproductive tract defects individually, would produce cumulative effects even when each chemical is present at a no observed adverse effect level (NOAEL) specific for male reproductive effects. Pregnant Sprague-Dawley rats were exposed via oral gavage to a fixed-ratio dilution mixture of 5 pesticides (vinclozolin, linuron, procymidone, prochloraz, pyrifluquinazon), 1 pesticide metabolite (dichlorodiphenyldichloroethylene (DDE)), and 9 phthalates (dipentyl, dicyclohexyl, di-2-ethylhexyl, dibutyl, benzyl butyl, diisobutyl, diisoheptyl, dihexyl, and diheptyl) during the critical window of rat fetal masculinization (gestation day 14–18). The top dose (100% dose) contained each compound at a concentration 2-fold greater than the individual chemical NOAEL followed by a dilution series that represented each chemical at NOAEL, NOAEL/2, NOAEL/4, NOAEL/8, NOAEL/15, NOAEL/100, NOAEL/1000. Reduced fetal testis gene expression occurred at NOAEL/15, reduced fetal testis testosterone production occurred at NOAEL/8, reduced anogenital distance, increased nipple retention, and delayed puberty occurred at NOAEL/4, and severe effects including genital malformations and weight reductions in numerous reproductive tissues occurred at NOAEL/2. This study demonstrates that these phthalates and pesticides acted cumulatively to produce adverse effects at doses below which any individual chemical had been shown to produce an effect alone and even though they have different MIEs. [ABSTRACT FROM AUTHOR]

• Selecting the right male for breeding is a prerequisite for maintaining a healthy and fertile herd. • An increase in the male reproductive disorders among animals prompts a search for new markers of male fertility. • Aquaporins are widely distributed in the male reproductive organs and sperm. • Aquaporins play a crucial role in the biology of male reproduction. • Possibility of using aquaporins for evaluation of male fertility and infertility in livestock and poultry. Improving the methods utilized to facilitate reproduction is associated with a constant need to search for new factors that not only significantly affect reproductive processes, but also create new possibilities when assessing male reproductive potential. Aquaporins (AQPs) belong to a family of small (28−30 kDa) proteins that facilitate the transport of water and other small molecules. There have been 13 AQPs (AQP0-AQP12) discovered in mammals, and these proteins are present in a wide range of cell types. Almost all AQPs, except AQP6 and AQP12 are present in the male reproductive organs and sperm of mammals and birds. Increasing evidence suggests that these proteins are involved in a number of processes responsible for the optimal functioning of the male reproductive system. This review presents the current state of knowledge regarding the abundance and distribution of AQPs in the male reproductive organs and sperm of various livestock and poultry species, including buffalo, cattle, sheep, horses, pigs, turkeys and goose. Furthermore, the possible physiological and pathophysiological significance of AQPs in male reproduction, as well as hormonal regulation of quantities are discussed. It can be concluded from the studies analyzed in this paper that abundance patterns of AQPs may be considered in the future as specific and universal biomarkers of male fertility and infertility in animal husbandry. [ABSTRACT FROM AUTHOR]

Extracellular vesicles (EVs) are small lipid bound structures released from cells containing bioactive cargoes. Both the type of cargo and amount loaded varies compared to that of the parent cell. The characterisation of EVs in cancers of the male urogenital tract has identified several cargoes with promising diagnostic and disease monitoring potential. EVs released by cancers of the male urogenital tract promote cell-to-cell communication, migration, cancer progression and manipulate the immune system promoting metastasis by evading the immune response. Their use as diagnostic biomarkers represents a new area of screening and disease detection, potentially reducing the need for invasive biopsies. Many validated EV cargoes have been found to have superior sensitivity and specificity than current diagnostic tools currently in use. The use of EVs to improve disease monitoring and develop novel therapeutics will enable clinicians to individualise patient management in the exciting era of personalised medicine. [ABSTRACT FROM AUTHOR]

Abstract: Polyarteritis nodosa (PAN) presents mostly as a systemic disease with poor prognosis, rarely in an isolated form with a usually favorable outcome. Both forms may affect the male reproductive system and both forms have been associated with malignancies. We describe for the first time the occurrence of isolated PAN in the reproductive system combined with a mixed germ cell tumor of the testis in a 21-year-old man presenting with symptoms of chronic epididymitis. Two years after surgery he is without evidence of recurrence of either the tumor or PAN. [Copyright &y& Elsevier]

Abstract: Prevailing studies emphasize on endocannabinoid activity in brain. However, sporadic evidences hint that endocannabinoid system controls male reproduction ranging from invertebrates to vertebrates. Although N-arachidonoylethanolamine is described in rat testis, its activity is still poorly known. Type-1 cannabinoid receptor and fatty acid amide hydrolase are particularly expressed in elongating spermatids and spermatozoa suggesting that endocannabinoids affect spermiogenesis and sperm physiology. Aim of this paper is to provide an analysis of the information available in vertebrates on male germ cell progression and sperm maturation mediated by the endocannabinoid system. [Copyright &y& Elsevier]