Salmon oil supplementation in dogs affects the blood flow of testicular arteries.

The administration of fish oils is known to cause changes in several reproductive parameters of domestic animals. The ingestion of polyunsaturated fatty acids (PUFAs) of the omega-3 family, such as docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), has been described and correlated with changes in the semen quality, testosterone levels and male fertility. Nevertheless, few studies monitored and registered effects after ceasing supplementation. In the present study, we monitored the Doppler velocimetric and ultrasonographic parameters of nine dogs' testis for 90 days (D90) checking the effect of salmon oil supplementation, and monitoring continued for 60 days more, after ceasing supplementation (D150). Ultrasonographic evaluations comprised determining the Doppler velocimetric parameters, testicular and epididymal volume, and testicular echotexture. Peak systolic velocity (PSV) as well as final diastolic velocity (EDV) in the supratesticular arteries (STA), and marginal artery (MA) increased during the period of treatment and kept that level up to D150. There was no difference between the fish-oil supplementation period and the unsupplemented one regarding the testicular and epididymal volume and echogenicity and heterogeneity characteristics. A negative correlation was found between heterogeneity of testis and sperm production (r = -.41, p = .008). Doppler velocimetry indices were affected by the supplementation, leading to an increase in testicular blood flow.

Seminal plasma protein networks and enriched functions in varicocele: Effect of smoking.

To verify a possible synergistic effect of smoking and varicocele on the seminal plasma proteome and biological functions, a cross-sectional study was performed in 25 smokers and 24 nonsmokers. Samples were used for conventional semen analysis, functional analysis (DNA fragmentation, acrosome integrity and mitochondrial activity) and proteomics by a shotgun approach. Functional enrichment of biological pathways was performed in differentially expressed proteins. Smokers presented lower ejaculate volume (p = .027), percentage of progressively motile spermatozoa (p = .002), total sperm count (p = .039), morphology (p = .001) and higher percentage of immotile spermatozoa (p = .03), round cell (p = .045) and neutrophil count (p = .009). Smokers also presented lower mitochondrial activity and acrosome integrity and higher DNA fragmentation. We identified and quantified 421 proteins in seminal plasma, of which one was exclusive, 21 were overexpressed and 70 were underexpressed in the seminal plasma of smokers. The proteins neprilysin, beta-defensin 106A and histone H4A were capable of predicting the smoker group. Enriched functions were related to immune function and sperm machinery in testis/epididymis. Based on our findings, we can conclude that cigarette smoking leads to the establishment of inflammatory protein pathways in the testis/epididymis in the presence of varicocele that seems to act in synergy with the toxic components of the cigarette.

Pre-cultivation of adipose tissue-derived microvascular fragments in porous scaffolds does not improve their in vivo vascularisation potential.

Adipose tissue-derived microvascular fragments represent promising vascularisation units for implanted tissue constructs. However, their reassembly into functional microvascular networks takes several days, during which the cells inside the implants are exposed to hypoxia. In the present study, we analysed whether this critical phase may be overcome by pre-cultivation of fragment-seeded scaffolds prior to their implantation. Green fluorescent protein (GFP)-positive microvascular fragments were isolated from epididymal fat pads of male C57BL/6-TgN (ACTB-EGFP) 1Osb/J mice. Nano-size hydroxyapatite particles/poly (ester-urethane) scaffolds were seeded with these fragments and cultivated for 28 days. Subsequently, these scaffolds or control scaffolds, which were freshly seeded with GFP-positive microvascular fragments, were implanted into the dorsal skinfold chamber of C57BL/6 wild-type mice to study their vascularisation and incorporation by means of intravital fluorescence microscopy, histology and immunohistochemistry over 2 weeks. Pre-cultivation of microvascular fragments resulted in the loss of their native vessel morphology. Accordingly, pre-cultivated scaffolds contained a network of individual CD31/GFP-positive endothelial cells with filigrane cell protuberances. After implantation into the dorsal skinfold chamber, these scaffolds exhibited an impaired vascularisation, as indicated by a significantly reduced functional microvessel density and lower fraction of GFP-positive microvessels in their centre when compared to freshly seeded control implants. This was associated with a deteriorated incorporation into the surrounding host tissue. These findings indicate that freshly isolated, non-cultivated microvascular fragments should be preferred as vascularisation units. This would also facilitate their use in clinical practice during intra-operative one-step procedures.

Cyclohexane-1,2-dicarboxylic acid diisononyl ester and metabolite effects on rat epididymal stromal vascular fraction differentiation of adipose tissue.

Plastics are generally mixed with additives like plasticizers to enhance their flexibility, pliability, and elasticity proprieties. Plasticizers are easily released into the environment and are absorbed mainly through ingestion, dermal contact, and inhalation. One of the main classes of plasticizers, phthalates, has been associated with endocrine and reproductive diseases. In 2002, 1,2-cyclohexane dicarboxylic acid diisononyl ester (DINCH) was introduced in the market for use in plastic materials and articles intended to come into contact with food, and it received final approval from the European Food Safety Authority in 2006. At present, there is limited knowledge about the safety and potential metabolic and endocrine-disrupting properties of DINCH and its metabolites. The purpose of this study was to evaluate the biological effects of DINCH and its active metabolites, cyclohexane-1,2-dicarboxylic acid (CHDA) and cyclohexane-1,2-dicarboxylic acid mono isononyl ester (MINCH), on rat primary stromal vascular fraction (SVF) of adipose tissue. DINCH and its metabolite, CHDA, were not able to directly affect SVF differentiation. However, exposure of SVF to 50 µM and 100 µM concentrations of MINCH affected the expression of Cebpa and Fabp4, thus inducing SVF preadipocytes to accumulate lipids and fully differentiate into mature adipocytes. The effect of MINCH was blocked by the specific peroxisome proliferator-activated receptor (PPAR)-α antagonist, GW6471. Taken together, these results suggest that MINCH is a potent PPAR-α agonist and a metabolic disruptor, capable of inducing SVF preadipocyte differentiation, that may interfere with the endocrine system in mammals.

15(S)-HETE-induced angiogenesis in adipose tissue is mediated through activation of PI3K/Akt/mTOR signaling pathway.

Chronic low-grade inflammation underlies obesity and associated metabolic dysfunctions. Lipoxygenase pathways are activated in adipose tissue during obese conditions. Since adipogenesis is associated with angiogenesis, the present study was designed to examine the role of 15-lipoxygenase metabolite, 15(S)-hydroxyeicosatetraenoic acid [15(S)-HETE] on angiogenesis in adipose tissue. Results showed that 15(S)-HETE induced sprouting in fat pad stromovascular tissues, induced morphological changes relevant to angiogenesis in endothelial cells derived from adipose tissue, upregulated the production of CD31, upregulated the gene level expression and production of vascular endothelial growth factor (VEGF), indicating the pro-angiogenic effect of 15(S)-HETE. LY294002, an inhibitor of PI3K-Akt pathway, and rapamycin, inhibitor of mammalian target of rapamycin (mTOR), significantly reversed the effect of 15(S)-HETE. 15(S)-HETE also induced activation of Akt and mTOR. These observations suggest that 15(S)-HETE stimulates angiogenesis in adipose tissue through activation of PI3K/Akt/mTOR signaling.

The blood-epididymis barrier and human male fertility.

Spermatozoa undergo a posttesticular maturation in the epididymis to acquire motility and the capacity to fertilize. Sperm maturation depends in part upon the creation of a specific microenvironment within the epididymal lumen. This environment is conditioned by proteins secreted by the epithelium and by exchange of molecules between the lumen and the blood circulation. These exchanges are selectively regulated by the blood-epididymis barrier. The blood-epididymis barrier is comprised of apical tight junctions between adjacent principal cells. Adherens junctions, which are necessary for cell adhesion, can also be found at the junctional complex present between adjacent principal cells. Progress has been made on the understanding of cellular interactions in the epididymis as well as the regulation of the luminal microenvironment and its importance for sperm maturation in rodents and humans. Clearly, changes in the function of cellular junctions in the human epididymis are associated with male infertility.

Ageing changes testes and epididymis blood flow without altering biometry and echodensity in dogs.

Senescence leads to deleterious effects in testicular function, sperm quality and fertility in dogs. There, however, are no consistent results of vascular changes in the testes and epididymis during natural ageing in dogs. The aim of this study, therefore, was to compare testes and epididymis blood flow, biometry and echodensity between young and senile dogs. Ten young dogs (1-4 years) and eight senile dogs (over 7 years) were selected and assigned to two experimental groups: Young Group and Senile Group. Dogs were evaluated using testicular and epididymis B-mode (dimensions and echodensity) and Doppler ultrasonography (tissue perfusion parameters and blood flow velocity of the testicular artery). There were no differences between experimental groups for the echographic evaluation of testicular and epididymis parenchyma and biometric variables. The dogs in the Young Group had greater (P = 0.02) testes vascularization score and greater (P = 0.06) testicular artery blood flow velocity than those in the Senile Group. Furthermore, the older dogs had a greater (P = 0.06) pulsatility index of the testicular artery than those in the Young Group. Ageing, therefore, seems to cause natural hemodynamical changes to the testicular artery, resulting in reduced blood flow (ischemia) and tissue damage. Testes and epididymis vascular characteristics, therefore, may represent the causal factors for changes in spermatogenesis and, as a consequence, negatively affect the sperm quality of older dogs. In conclusion, senescence alters testicular artery blood flow and vascularization of the testes, without changing testicular and epididymis ultrasonographic dimensions and echodensity in dogs.

Angiogenic potential of microvessel fragments is independent of the tissue of origin and can be influenced by the cellular composition of the implants.

UNLABELLED: We have demonstrated that MFs isolated from adipose retain angiogenic potential in vitro and form a mature, perfused network when implanted. However, adipose-derived microvessels are rich in provascularizing cells that could uniquely drive neovascularization in adipose-derived MFs implants. OBJECTIVE: Investigate the ability of MFs from a different vascular bed to recapitulate adipose-derived microvessel angiogenesis and network formation and analyze adipose-derived vessel plasticity by assessing whether vessel function could be modulated by astrocyte-like cells. METHODS: MFs were isolated by limited collagenase digestion from rodent brain or adipose and assembled into 3D collagen gels in the presence or absence of GRPs. The resulting neovasculatures that formed following implantation were assessed by measuring 3D vascularity and vessel permeability to small and large molecular tracers. RESULTS: Similar to adipose-derived MFs, brain-derived MFs can sprout and form a perfused neovascular network when implanted. Furthermore, when co-implanted in the constructs, GRPs caused adipose-derived vessels to express the brain endothelial marker glucose transporter-1 and to significantly reduce microvessel permeability. CONCLUSION: Neovascularization involving isolated microvessel elements is independent of the tissue origin and degree of vessel specialization. In addition, adipose-derived vessels have the ability to respond to environmental signals and change vessel characteristics.

Difference in abundance of blood and lymphatic capillaries in the murine epididymis.

It is well known that the epididymis is responsible for the transportation, storage, and maturation of spermatozoa, and that it has four segments: i.e., initial segment, caput, corpus, and cauda. Previous studies revealed the presence of a very dense network of blood capillaries only in the initial segment; however, the relative distribution of blood and lymphatic capillaries has remained unknown. In the present study, we investigated the distribution of intraepididymal blood and lymphatic capillaries in mice by use of monoclonal antibodies against CD31 and lymph vessel endothelium HA-receptor 1, respectively. The results showed that lymphatic networks were quite scarce in the initial segment and strikingly abundant in the cauda compared with the other two regions. In sharp contrast, blood capillaries were abundant in both the initial segment and cauda but sparse in the other two regions. This observation suggests that the characteristic distribution of blood and lymphatic capillaries in the epididymis contributes to the different roles of each epididymal region.

[Case of isolated polyarteritis nodosa appearing in bilateral epididymis].

A 50-year-old man visited our hospital with a complaint of painful masses in bilateral scrotums for 2 months. Ultra sonography shows isoechoic mass in both scrotum. Antibiotic was treated for 2 weeks but had no effects on scrotal swelling and testicular neoplasm cannot ruled out, so we performed bilateral high orchiectomy. Pathological examination shows necrotizing vasculaitis surrounded by glanulomas and pathologically diagnosed as Polyarteritis Nodosa in bilateral epididymis. After orchiectomy, blood examination of immunity was revealed no specific findings, that made us diagnosed as isolated PN. 2 years after operation, he had no reccurence.

Arterial supply to the rabbit male genital organs.

The improvement of veterinary care has prolonged the lifespan of rabbits, and the number of rabbits suffering from age-related, male genital disorders may increase in the near future. This could result in increased opportunities for male genital surgery, requiring knowledge of their arterial anatomy, which, however, has not been sufficiently studied. Therefore, the arteries supplying the genitals were observed in 20 male New Zealand White rabbits. The testis was supplied by the testicular artery originating from the abdominal aorta. The right testicular artery usually emerged at a more cranial level than the left artery (65%). The testicular artery encircled the testis in the sagittal plane and bifurcated (95%) or trifurcated (5%) at the caudal extremity of the testis before entering the parenchyma. The epididymis was supplied by the epididymal branches, either from only the testicular artery (75% of the right and 80% of the left halves) or from both the testicular artery and aorta. The deferent duct was supplied in all halves by the dorsal and ventral branches of the deferential artery, which usually arose from the umbilical artery. The accessory genital glands were supplied by the dorsal branch of the deferential artery and the prostatic artery. The latter, which emerged from the internal iliac artery, exhibited 3 branching types. The most frequent type (55% of the right and 45% of the left halves) had 3 branches supplying the accessory genital glands. These findings will help improve rabbit genital surgery.

Ultrasonographic characteristics of the testes, epididymis and accessory sex glands and arterial spectral indices in peri- and post-pubertal Nelore and Caracu bulls.

Ultrasonography can provide information about the integrity of organs; however, rarely is applied to the reproductive organ evaluation of bulls. The objective of the present study was to characterize and compare values for variables and ultrasonographic characteristics of the testes, epididymis and accessory sex glands, as well as spectral Doppler indices of the testicular and internal iliac arteries, between peri- and post-pubertal Nelore and Caracu bulls. Nelore (n = 203) and Caracu (n = 79) bulls were assigned by age class: peri-pubertal (12-15 months) and post-pubertal (> 22 months). Data were analyzed using SAS's PROC MIXED procedure (P < 0.05). The biometric variables of the testes and cauda epididymis differed between peri- and post-pubertal Nelore and Caracu bulls. There was a difference between breeds for the vesicular glands, ampulla of vas deferens, disseminate portion of the prostate, and craniocaudal dimension of the bulbourethral glands. Echogenicity of the testicular parenchyma differed between breeds and age classes. The pulsatility and resistive indices of the testicular arteries differed between Nelore and Caracu bulls. The biometric and ultrasonographic characteristics of the testes, epididymis and accessory sex glands, as well as of the arterial indices in bulls are affected by genetic group and age class, and when assessed there is useful information regarding the progression of sexual maturation.

Overexpression of VEGF in testis and epididymis causes infertility in transgenic mice: evidence for nonendothelial targets for VEGF.

Vascular endothelial growth factor (VEGF) is a key regulator of endothelial growth and permeability. However, VEGF may also target nonendothelial cells, as VEGF receptors and responsiveness have been detected for example in monocytes, and high concentrations of VEGF have been reported in human semen. In this work we present evidence that overexpression of VEGF in the testis and epididymis of transgenic mice under the mouse mammary tumor virus (MMTV) LTR promoter causes infertility. The testes of the transgenic mice exhibited spermatogenic arrest and increased capillary density. The ductus epididymidis was dilated, containing areas of epithelial hyperplasia. The number of subepithelial capillaries in the epididymis was also increased and these vessels were highly permeable as judged by the detection of extravasated fibrinogen products. Intriguingly, the expression of VEGF receptor-1 (VEGFR-1) was detected in certain spermatogenic cells in addition to vascular endothelium, and both VEGFR-1 and VEGFR-2 were also found in the Leydig cells of the testis. The infertility of the MMTV-VEGF male mice could thus result from VEGF acting on both endothelial and nonendothelial cells of the male genital tract. Taken together, these findings suggest that the VEGF transgene has nonendothelial target cells in the testis and that VEGF may regulate male fertility.

Angiogenic role of LYVE-1-positive macrophages in adipose tissue.

Here we report the discovery of a characteristic dense vascular network (DVN) in the tip portion of epididymal adipose tissue in adult mice. The DVN is formed by angiogenesis rather than by vasculogenesis, and has functional blood circulation. This DVN and its subsequent branching may provide a new functional route for adipogenesis. The recruitment, infiltration, and accumulation of bone marrow-derived LYVE-1(+) macrophages in the tip region are crucial for the formation of the DVN. Matrix metalloproteinases (MMPs) and the VEGF-VEGFR2 system are responsible not only for the formation of the DVN, but also for the recruitment and infiltration of LYVE-1(+) macrophages into the epididymal adipose tissue tip region. SDF-1, but not the MCP-1-CCR2 system, is a critical factor in recruitment and ongoing retention of macrophages in this area. We also demonstrate that the tip region of epididymal adipose tissue is highly hypoxic, and thus provides a microenvironment conducive to the high expression and enhanced activities of VEGF, VEGFR2, MMPs, and SDF-1 in autocrine and paracrine manners, to create an ideal niche for the recruitment, retention, and angiogenic action of macrophages. These findings shed light on the complex interplay between macrophage infiltration, angiogenesis, and adipogenesis in the tip region of adult epididymal adipose tissue, and provide novel insight into the regulation of alternative outgrowth of adipose tissue.

Of vessels and cells: the spatial organization of the epididymal immune system.

BACKGROUND: One third of infertility cases in couples worldwide has an exclusive male origin and immune disorders, essentially due to repetitive infections, are emerging an cause of male infertility. As the place of sperm maturation, epididymis must be preserved from excessive immune responses that may arise following infections of the male genital tract. At the same time, epididymis must set and maintain a tolerogenic environment in order not to destroy sperm cells that enter the tissue at puberty, long after the immune system has been taught to recognize self pathogens. The immune cells that populate the epididymis have raised growing interest over the last thirty years but they may be not sufficient to understand the immune balance existing in this organ, between immune response to pathogens and tolerance to spermatozoa. Indeed, immune cells are the most motile cells in the organism and need blood and lymphatic vessels to traffic between lymphoid organs and sites of infection to induce efficient responses. OBJECTIVES: To review the literature on the blood and lymphatic vessels, and on the immune cells present at steady state in the rodent epididymis (rat and mouse). MATERIALS AND METHODS: PubMed database was searched for studies reporting on the spatial organization of the rodent epididymal vasculature and immune cell types at steady state. This search was combined with recent findings from our team. RESULTS: At steady state, the rodent epididymis presents with dense blood and lymphatic networks, and a large panel of immune cells distributed across the interstitum and epithelium along the organ. CONCLUSIONS: The immune system of the rodent epididymis is highly organized. Exploring its functions, especially in an infectious context, is the essential coming step before any transposition to human.

Macroscopic features of the venous drainage of the reproductive system of the male ostrich (Struthio camelus).

The macroscopic features of the venous drainage of the reproductive system of the male ostrich were studied in six pre-pubertal and three sexually mature and active birds. Each testis was drained by one to four testicular veins. The right testicular veins drained the right testis and epididymis and its appendix to the caudal vena cava and to the right common iliac vein, whereas the left testicular veins drained the left testis and epididymis and its appendix exclusively to the left common iliac vein. A number of variations in the drainage pattern based on the point of entry and number of testicular veins were observed. The cranial aspect of the testis was also linked to the caudal vena cava or common iliac vein via the adrenal veins. The cranial, middle and caudal segments of the ductus deferens (and ureter) were drained by the cranial, middle and caudal ureterodeferential veins respectively, to the caudal testicular veins, the caudal renal veins and pudendal/caudal part of the internal iliac veins. In some specimens, the caudal ureterodeferential veins also drained into the caudal mesenteric vein. The surface of the phallus was drained by tributaries of the pudendal vein. The basic pattern of venous drainage of the reproductive organs of the male ostrich was generally similar to that described for the domestic fowl. However, important differences, including the partial fusion of the caudal renal veins, drainage of the cranial aspect of the testes via the adrenal veins, drainage of the caudal ureterodeferential veins into the caudal mesenteric vein and the presence of veins draining the surface of the phallus, were observed. Although significant, these differences may simply reflect variations in the normal pattern of venous drainage of the reproductive tract of birds which could be verified by studying more specimens and more species.

The role of cell adhesion molecules in ischemic epididymal injury.

OBJECTIVE: The aim of this study was to investigate the role of adhesion molecules in epididymal injury induced by ischemia-reperfusion (I-R) in the rats. STUDY DESIGN: About 20 male Sprague-Dawley rats were separated into two groups. A sham operation was performed in group 1 (control). In group 2 (I-R), following 6 h of unilateral spermatic cord torsion, 1-h detorsion of the testis was performed. Then, epididymides were removed to measure the tissue levels of malondialdehyde (MDA) and to make histological examination. RESULTS: Malondialdehyde values increased in group 2. In group 2, the rats demonstrated significant disorganization of the epithelium and loss of microvilli in the epididymal tissue. No abnormal microscopic findings of the epididymis of the rats in the control group. The tenascin expression in the interstitial area of the epididymis was intense in group 2. Intercellular adhesion molecule-1 (ICAM-1) expression by intense brown staining was seen along the basement membrane in epididymal tissue from I-R group rats. The microvillus sites of the epithelia in I-R group were stained mildly by lectin. CONCLUSION: The increased expression of adhesion molecules found in epididymal injury induced during of postischemic reperfusion may implicate importance of inflammatory infiltration.

Nestin in the epididymis is expressed in vascular wall cells and is regulated during postnatal development and in case of testosterone deficiency.

Vascular smooth muscle cells (SMCs), distinguished by the expression of the neuronal stem cell marker nestin, may represent stem cell-like progenitor cells in various organs including the testis. We investigated epididymal tissues of adult nestin-GFP mice, rats after Leydig cell depletion via ethane dimethane sulfonate (EDS), rats and mice during postnatal development and human tissues. By use of Clarity, a histochemical method to illustrate a three-dimensional picture, we could demonstrate nestin-GFP positive cells within the vascular network. We localized nestin in the epididymis in proliferating vascular SMCs by colocalization with both smooth muscle actin and PCNA, and it was distinct from CD31-positive endothelial cells. The same nestin localization was found in the human epididymis. However, nestin was not found in SMCs of the epididymal duct. Nestin expression is high during postnatal development of mouse and rat and down-regulated towards adulthood when testosterone levels increase. Nestin increases dramatically in rats after Leydig cell ablation with EDS and subsequently low testosterone levels. Interestingly, during this period, the expression of androgen receptor in the epididymis is low and increases until nestin reaches normal levels of adulthood. Here we show that nestin, a common marker for neuronal stem cells, is also expressed in the vasculature of the epididymis. Our results give new insights into the yet underestimated role of proliferating nestin-expressing vascular SMCs during postnatal development and repair of the epididymis.