Testicular Capillaries Research Articles

Effect of feeding corn silage on semen quality and spermatogenesis of bulls.

Bovine reproduction, including male fertility traits like semen quality, are influenced by a variety of different factors like breed, nutrition, environment, and feeding management. Diet in a crucial determinant, and in this regard although corn silage is generally considered to be a favorable roughage for fattening meat type breeds, it tends to have a negative impact on semen quality. In the current study, alfalfa hay was substituted by corn silage as a roughage source in the diet of bulls to investigate its effects on the fertility of breeding bulls. A feeding trail spanning 140 days was conducted, with semen collection occurring twice a week commencing 60 days after the start of trial. Semen quality parameters, serum antioxidant indexes, sex hormone content in semen, rumen microflora, and sperm transcriptome were characterized. Feeding corn silage enhanced host antioxidant capacity, significantly decreased spermatozoal motility and increased sperm deformity rate in bulls. Furthermore, follicle-stimulating hormone (FSH) and luteinizing hormone (LH) content in semen were significantly decreased (P < 0.05), and the inhibin B (INHB) content was significantly increased (P < 0.01). Feeding corn silage led to significant changes in the diversity of rumen microbiota of cattle at the phylum and genus levels, some of which were significantly correlated with semen quality. Subsequent RNA sequencing indicated that DHH and PITHD1, two genes related to sperm and reproductive development, were differentially expressed, and enrichment analysis also identified several pathways and biological functions relevant to sperm development and reproduction. These results indicate that feeding corn silage modulates semen quality via different pathways. Firstly, corn silage metabolites likely affect the secretion of INHB through the testicular capillaries, which affects semen quality by regulating genes involved in spermatogenesis. Secondly, low lignin content in silage corn appears to reduce abundance of rumen flora that are positively correlated with semen quality. Overall, results indicate that feeding bulls corn silage as the primary source of forage could negatively impact semen quality and may not be appropriate as the primary roughage of forage for breeding bulls.

Low testosterone in ApoE/LDL receptor double-knockout mice is associated with rarefied testicular capillaries together with fewer and smaller Leydig cells

The testis as a site for atherosclerotic changes has so far attracted little attention. We used the apolipoprotein E (ApoE)/low density lipoprotein (LDL) receptor deficient mouse model (KO) for atherosclerosis (20, 40, 60 and 87-week-old) to investigate whether Leydig cells or the capillary network are responsible for reduced serum testosterone levels previously observed in extreme ages of this model. In KO mice, overall testosterone levels were reduced whereas the adrenal gland-specific corticosterone was increased excluding a general defect of steroid hormone production. In addition to micro-CT investigations for bigger vessels, stereology revealed a reduction of capillary length, volume and surface area suggesting capillary rarefaction as a factor for diminished testosterone. Stereological analyses of interstitial cells demonstrated significantly reduced Leydig cell numbers and size. These structural changes in the testis occurred on an inflammatory background revealed by qPCR. Reduced litter size of the KO mice suggests hypo- or infertility as a consequence of the testicular defects. Our data suggest reduced testosterone levels in this atherosclerosis model might be explained by both, rarefication of the capillary network and reduced Leydig cell number and size. Thus, this study calls for specific treatment of male infertility induced by microvascular damage through hypercholesterolemia and atherosclerosis.

Experimental study on contrast-enhanced ultrasound to evaluate testicular microvascular injury

Objective To explore the way of contrast-enhanced ultrasound(CEUS) and quantitative analysis technology to assess testicular microvascular ischemia-reperfusion injury. Methods Twenty-four healthy adult rabbits were randomly divided into the control group and the microvascular mild, moderate, serve ischemic injury group (group Ⅰ, including group Ⅰ1, Ⅰ2, Ⅰ3). The spermatic cord of the control group were threaded but not ligated. The group Ⅰ1, Ⅰ2, Ⅰ3 were reperfused 2 h after 2 h, 4 h, 6 h complete testicular ischemia respectively. The ultrasound contrast angiographic parameters of each group were analyzed by the quantitative analysis techniques, including peak intensity(PI), time to peak(TP), slope, mean transmit time(MTT), DT/2 and area. The levels of plasma vWF, NO and ET-1 were tested after the CEUS. The pathological changes of the ipsilateral testicular were observed by the HE and immunohistochemical staining. Statistical analysis was performed by SPSS13.0 software. Results Preoperativeiy, each parameter of the experimental group and the control group showed no statistical significance (P>0.05). Postoperatively, the PI, slope, area of the group Ⅰ1 increased significantly, while the TP, MTT were prolonged(P<0.05). Contrast time-intensity curve was significantly higher peak and retrusion. The group Ⅰ2′s PI, slope, area increased further, DT/2 was significantly prolonged, but TP significantly reduced(P<0.05). The contrast curve significantly increased, forward and decreased slowly. The contrast curves of the group Ⅰ3 was approximate a straight line. The vWF, NO, ET-1 levels of each experimental group were significantly higher than those in control group (P<0.05), among them, the levels of group Ⅰ2 were the highest(P<0.05). With the aggravation of ischemia, the testicular capillaries expansion, erythrocyte sedimentation, leakage, infiltration of inflammatory cells, MMP-9 expression in vascular basement membrane gradually increased. Conclusions The CEUS and quantitative analysis techniques can effectively evaluate testicular microvascular injury. Key words: Ultrasonography; Microbubbles; Testis; Ischemia; Reperfusion injury

The changes in testicular vascular permeability during progression of the experimental varicocele.

The vascular permeability of testicular capillaries which play a role in controlling the formation of testicular interstitial fluid was studied during the progressive course of experimental varicocele. The pathology was developed through partial ligation of left renal vein in four groups of rats. Controls of each group were subjected to sham surgery. After different periods of varicocele creation (1, 3, 6 and 14 weeks), animals' testes per one of the study groups were extirpated and weighed. The volume density percentages of polymorphnuclear leukocytes (PMN) per testicular blood vessels; which are markers of the increase in vascular permeability, were also estimated in both testes. To further verify the obtained findings, another group of animals received human chorionic gonadotrophin (hCG) treatment 6 weeks after varicocele creation and their histopathological sections were examined. Animal testes' of (1 and 3 weeks) groups were found to be significantly heavier (p<0.05) than their controls. PMN showed accumulation in testicular blood vessels and their volume density percentages per these blood vessels in both testes were significantly higher in each study group than in those of its controls. However, these percentages showed gradual significant decline as the duration of varicocele bearing gradually increased. The hCG-treated animals revealed more accumulation of the PMN in their histopathological sections. The present results suggest that experimental varicocele may induce an increase in testicular vascular permeability, which then decreases gradually with time. It is supposed that parallel changes in the rate of formation of testicular interstitial fluid may be accompanied. The results have also showed that the vasculature of the testis with experimental varicocele can still respond to hCG.

Morphologic and histochemical study of blood capillaries in boar testes: effects of abdominal cryptorchidism.

Few data exist about the features of testicular microvasculature under normal and pathologic conditions. The morphology and lectin affinity of testicular capillaries were examined in healthy boars and in unilateral and bilateral abdominal cryptorchid boars. The capillaries of scrotal testes contained a) the endothelial layer formed by two cells, b) the basal lamina constituted by collagen fibers and glycoconjugates with fucosyl, galactosyl, glucosyl, and neuraminic acid residues, and c) the pericyte layer formed by a single cell. These components participated in substrate exchange between blood and testicular tissue. The abdominal testes showed increased numbers of capillaries, which could exhibit a mature appearance, but also angiogenic or degenerative patterns. Angiogenesis was manifested in interstitial capillaries and was characterized by a) proliferation of endothelial cells, b) decreased thickness and decreased content of collagen fibers and glycoconjugates in the basal lamina, and c) lack of pericytes. Degenerative capillaries lay in association with seminiferous tubules and showed a) pyknotic endothelial cells; b) thickening, collagenization, and altered glycoconjugate content in the basal lamina; and c) increased development of pericytes. The angiogenesis of interstitial capillaries resulted in high vascular permeability, and the degeneration of intertubular capillaries led to defective substrate exchange between blood and seminiferous tubules. Unilateral abdominal cryptorchidism did not alter the morphology and function of capillaries in the scrotal testis. Unilateral and bilateral abdominal cryptorchidism resulted in increased numbers and abnormal morphology and function of capillaries in abdominal testes. The proliferation of interstitial capillaries correlated with the immaturity of Leydig cells, and the degeneration of intertubular capillaries correlated with the thickening of the lamina propria.

Endothelin and endothelin receptors A and B in the human testis.

Human testicular capillaries interconnect Leydig cells and seminiferous tubules. Microcirculation and blood flow are therefore essential for the maintenance of spermatogenesis. The expression and the localisation of ET (endothelin) and its receptors in testicular tissue, in seminiferous tubules and in human testicular capillaries were studied. ET-1 mRNA was detected in whole testicular tissue and in seminiferous tubules whereas isolated testicular capillaries were negative. Big ET-1 (Big endothelin 1) and ET peptides were localised in Leydig and Sertoli cells whereas interstitial and intramural capillaries (within the lamina propria) remained unstained. ET was also found in mature spermatids. ET-A (endothelin receptor A) mRNA was detected in seminiferous tubules and whole testicular tissue whereas testicular blood vessels were negative. ET-A immunostaining was displayed in Leydig and Sertoli cells and in spermatids. ET-B (endothelin receptor B) mRNA was detected in whole testicular tissue, seminiferous tubules and in testicular capillaries. ET-B peptide was prominent in Leydig cells, peritubular cells, endothelial cells and pericytes of interstitial and intramural capillaries as well as in vascular endothelial and smooth muscle cells. From these results we conclude that ET produced in Leydig and Sertoli cells can act in a paracrine manner via ET-B on the human testicular microvasculature and the peritubular cells. The presence of both ET-A and ET-B in Leydig cells and of ET-A in Sertoli cells leads to the assumption that ET could influence these cells as an autocrine factor.

Microcirculation and the vascular control of the testis.

The arterial blood supply of the human testicular parenchyma within a testicular lobule demonstrates a segmental organization. In correlation to Leydig cells and seminiferous tubules within such a testicular tissue segment the capillary pathway was subdivided into three parts, namely arterial side inter-Leydig cell capillaries, intramural capillaries and venous side inter-Leydig cell capillaries. The organization, ultrastructure and functional aspects of the human testicular microvasculature were studied in detail. The casting preparation demonstrated a tightly organized vascular network. Computer aided 3-D reconstructions revealed that the capillary pathway in the human testis interconnects Leydig cells and seminiferous tubules. Electron microscopically the endothelial cells (EC) of arterial- and venous side inter-Leydig cell capillaries as well as of intertubular capillaries free of Leydig cells were of the continuous type without fenestrations. The intramural capillaries consisted of non-fenestrated and fenestrated sections. In all cases studied, the fenestrations faced the germinal epithelium. Transcytotic vesicles were numerous in the EC of inter-Leydig cell capillaries and in the non-fenestrated part of intramural capillaries. Leydig cells and Sertoli cells demonstrated immunostaining for vascular endothelial growth factor (VEGF) and its receptors flt-1 and KDR. In agreement with data obtained by RTPCR analyses, human testicular capillaries were negative for VEGF but positive for its receptors. Immunohistochemically Leydig cells and Sertoli cells were also positive for Big-Endothelin and Endothelin-1. Endothelin receptors ЕТ-А and ET-B were localized in Leydig cells while Sertoli cells showed only ЕТ-А immunostaining. Testicular blood vessels and peritubular cells were only positive for ET-B. Androgen receptors could be localized on the arterial side of the human testicular microvasculature and on intramural capillaries, whereas the intralobular veins were negative. In contrast, estrogen receptors were found in all parts of testicular vasculature. From these results we conclude: 1) In the human testis, capillaries interconnect Leydig cell clusters and seminiferous tubules in a serial manner. A part of the capillaries runs in between the layers of the lamina propria and therefore represents the capillarization of human seminiferous tubules. It was presumed that the multilayered lamina propria requires its own capillary supply which may allow a rapid exchange of hormones and other nutritious substances between the microvasculature and the germinal epithelium. 2) VEGF produced and released from Leydig and Sertoli cells could act as a paracrine factor on the testicular microvasculature which possesses VEGF receptors and could modulate the permeability of capillaries in the adult testis, e.g. by fenestration of intramural capillaries. 3) The presence of androgen receptors (AR) and estrogen receptors (ER) on the testicular vasculature indicate that the testicular microvasculature is involved in the local endocrine and paracrine regulation of spermatogenesis. Particularly the presence of AR on the arterial side and on intramural capillaries leads to the assumption that androgens could influence the blood supply to Leydig cells and seminiferous tubules. 4) Leydig cells and Sertoli cells could also influence the blood flow via the potent vasoconstrictor ET-1.

Ultrastructural Characteristics of the Testicular Capillaries in the Dog (Canis familiaris, L.)

The main ultrastructural and cytophysiological characteristics observed on the testicular capillaries in the dog concern exocytosis and endocytosis processes, probably related to fluid and solute transport through the cells.

Microvascular pressure distribution in the hamster testis

Convective transport is a critical element in the regulation of steroidogenesis and spermatogenesis in the testis. Steroid hormones are distributed to their target cells within seminiferous tubules via interstitial fluid. The movement of interstitial fluid and lymph, which transports protein hormones and many of the substrates required for spermatogenesis and steroidogenesis, is driven by capillary filtration. Despite the importance of convective transport in testicular function, however, the mechanisms regulating transvascular exchange in the testis are unknown. As a first step in understanding this process, we measured directly the microvascular hydrostatic pressure distribution in the hamster testis (pentobarbital sodium, 70 mg/kg ip). Using a servo-null transducer, intravascular pressure was measured in all vessel types accessible beneath the surface of the testis of 19 animals. Systemic arterial pressure averaged 89 +/- 2 (SE) mmHg. The most significant observations were that mean capillary pressure was extremely low (10.1 +/- 0.8 mmHg) and remarkably constant (range 8.2-13.3 mmHg), despite a 45 mmHg range in systemic mean arterial pressure among the animals observed. The maintenance of a low hydrostatic pressure in testicular capillaries may serve to sustain fluid filtration at a rate that prevents washout of essential solutes while preserving convective transport. Unfortunately, the anatomical and functional characteristics that determine this unique microvascular environment may also expose the testis to significant pathological risks. For example, the large pre- to postcapillary resistance ratio observed suggests that testicular capillaries must be highly susceptible to increases in venous pressure.(ABSTRACT TRUNCATED AT 250 WORDS)

Ablation of the blood-testis barrier in rats and guinea pigs by 48/80, a histamine releaser, and cadmium chloride.

When adult male guinea pigs were injected unilaterally intratesticularly with compound 48/80 (1 mg in 0.05 ml saline), their testicular peritubular capillaries were engorged with blood, intratubular edema was present on the ipsilateral side, and the blood-testicular (BT) barrier (measured by the entrance of acriflavin into the seminiferous tubule) was ablated. The contralateral testis, when injected with saline, showed no pathological changes nor a breakdown of the BT barrier. Subcutaneous injection of guinea pigs with cadmium chloride resulted in a more intense intratubular fluorescence than was observed for 48/80-treated animals. The H1 and H2 receptor blockers (diphenhydramine and cimetidine, respectively) reduced the intensity of capillary engorgement and edema. Intraperitoneal injections of 48/80 (0.5 mg) was lethal to rats, and both diphenhydramine and cimetidine induced survival even with as much as 1 mg of 48/80. Intratubular fluorescence was less severe than that observed for similarly treated guinea pigs or from cadmium chloride-treated rat testes. Neither treatment (48/80 or cadmium chloride) altered the blood-epididymal barrier of either guinea pigs or rats. Species differences were observed in the lethal effects of exogenously administered histamine.

Testicular interstitial fluid as a monitor for changes in the intratesticular environment in the rat.

The use of testicular interstitial fluid (IF) collected from the rat testis has been validated as (1) an index of the total extracellular extratubular fluid volume of the testis (which reflects the permeability of testicular capillaries) and (2) as a means of measuring changes in the interstitial hormonal environment. The former was tested by comparing the albumin 'space' with the volume of recovered IF in the same testes from control and bilaterally cryptorchid rats sampled at 0-40 h after injection of hCG. Although the volume of IF recovered was on average only 50% of the albumin 'space', both measures increased in parallel after hCG injection and were always closely correlated (P less than 0.001) over a 4- to 5-fold range. The volume of recovered IF increased with age in parallel with increase in testicular weight, and the testosterone concentration in IF paralleled changes in peripheral serum, increasing from 45 to 80 days of age and then declining. After injection of 25 micrograms bovine LH, testosterone levels in IF, spermatic venous (SV) and peripheral venous (PV) blood increased up to 10-fold by 1 h and returned to control levels over the next 11 h. Testosterone levels in IF were always considerably higher than those in SV blood, but this difference was not constant. Subcutaneous injection of rats with an LH-RH agonist resulted in parallel increases in the serum levels of LH and in the IF and PV levels of testosterone. However, at 6 h there was an 'LH-independent' secondary increase in testosterone levels which was associated with an increase in IF volume, reflecting an increase in capillary wall permeability and hence increased transport of LH into the testis.

Impaired gonadotrophin uptake in vivo by the cryptorchid rat testis.

The specific testicular uptake in vivo of 125I-labelled hCG was compared in control adult rats and adult rats made bilaterally cryptorchid 5 weeks previously. Although a similar temporal pattern of uptake was observed in both groups, uptake of hCG by cryptorchid testes was reduced at all times after injection by up to 70%. The possible causes of this impairment were investigated. It could not be accounted for by differences in the rate of absorption or clearance of 125I-labelled hCG in the two groups. Therefore, because hCG-induced increase in the permeability of testicular capillaries is a crucial factor in determining hCG uptake by the testis, this change was compared in control and cryptorchid testes. Although hCG induced a characteristic increase in testicular capillary wall permeability in both groups, this change was temporally delayed in cryptorchid testes, and occurred after hCG values in the blood had fallen. Even when hCG had crossed the capillary wall into testicular interstitial fluid, its uptake into the testicular tissue was significantly lower in cryptorchid than in control testes. These changes probably account for the impairment of gonadotrophin uptake by the cryptorchid testis and have important implications with respect to the aetiology of Leydig cell changes in cryptorchidism.

Horseradish peroxidase as a tracer for capillary permeability studies.

The permeability of capillaries was investigated utilizing an in vivo injection of horseradish peroxidase (HRP) and an in situ perfusion of a balanced salt solution containing HRP and lanthanum chloride. In the continuous capillaries of heart and muscle, HRP diffuses mainly through intercellular junctions, while in testicular capillaries, the transport is via micropinocytotic vesicles. The diffusion and micropinocytotic transport of HRP was demonstrated in both directions, i.e. from the capillary lumen to the interstitium and vice versa. Lanthanum can be used as a bidirectional inhibitor of micropinocytosis. The transport of HRP is then almost completely hindered in testicular capillaries. In heart muscle, the effect on HRP transport is not significant, due to second transport pathway, i.e. intercellular cleft passage.

The leprous testis.

A clinical investigative study of 148 male leprous patients demonstrated the presence of testicular lesions in 35 cases. Semen analysis revealed marked oligo-athenozoospermia in 10 cases and azoospermia in 25 cases. Testicular biopsies from leprous testes showed different histologic patterns ranging from spermatogenic arrest to complete hyalinization of both seminiferous tubules and interstitial tissue. Histochemical staining for neurovascular supply revealed degenerative nerve change in addition to altered permeability of the testicular capillaries. There was good correlation between the results of semen analysis and histological and histochemical examination of testicular biopsies.

Reexamination of the Lesions in Rat Testis Caused by Cadmium

The early changes in the microvascular bed of the testis resulting from treatment with cadmium were studied with ti-ypan blue, electron dense tracers and by vascular injections. Leakage of fluids and electrolytes from the testicular blood vessels to the interstitium was demonstrated at the light and electron microscope levels as early as 1-2 h following parenteral injection of cadmium. After 1-2 h, discontinuities in the endothelial lining could be detected by the presence of carbon particles in the walls of the testicular capillaries and venules. By 3 h, the extravasation of carbon had increased, such that particles could be seen labeling large veins. Within the vessels, a local increase in the concentration of erythrocytes was noted and numerous platelets adhering to exposed subendothelial structures were seen plugging the endothelial gaps. At 4 h, a striking decrease in labeling of veins was noted due to the fact that delivery of the tracer towards the venous system is almost entirely obstructed as a result of the blockage and progressive deterioration of the smaller vessels; ischemia of the testis ensued from the obstruction of the microvascular circulation. These results indicate that the degeneration of the seminiferous epithelium and all biochemical and physiological changes known to occur in the testis at later time intervals following cadmium treatment are secondary to ischemia rather than due to a direct effect of the cadmium.

Dye permeability and alkaline phosphatase activity of testicular capillaries in the postnatal rat.

Staining of testicular and epididymal tissues after intravenous, intraperitoneal or subcutaneous administration of a number of dyes was investigated in rats at different stages of postnatal development. After light green injections heavy staining of both testis and epididymis was visible to the naked eye in neonatal animals up to the age of 10 days, while in rats over 15 days old no appreciable staining of the testis could be seen, although the caput epididymis was strongly coloured. From 3–8 hours after subcutaneous acriflavine administration, the nuclei in the blood vessel walls of the testis, as well as the nuclei in the rete testis, tubuli efferentes and caput epididymis, fluoresced in all age groups. The nuclei of the interstitial and tubular cells were stained intensely until the age of 5 days. Thereafter the intensity gradually diminished until the age of 20 days, when no nuclear fluorescence was visible in the seminiferous tubules and even the interstitial nuclei fluoresced weakly or not at all. The histochemical alkaline phosphatase activity of the testicular capillaries was studied by Gomori's method, using fresh and postfixed cryostat sections from postnatal rat testes. The testicular capillaries exhibited appreciable activity at the age of 10 days. On the basis of the present and previous observations on the permeability of the testicular capillaries, the existence of a blood-testis barrier in the puberal and adult rat testis is suggested. Development of the blood-testis barrier and the alkaline phosphatase activity of the testicular capillaries are suggested to reflect general vascular maturation at the beginning of puberty in the rat.

Endocrine gland blood flow in the unanesthetized, unrestrained rat

SPECIAL COMMUNICATIONSEndocrine gland blood flow in the unanesthetized, unrestrained ratHarold GoldmanHarold GoldmanPublished Online:01 Jul 1961https://doi.org/10.1152/jappl.1961.16.4.762MoreSectionsPDF (363 KB)Download PDF ToolsExport citationAdd to favoritesGet permissionsTrack citations ShareShare onFacebookTwitterLinkedInWeChat Previous Back to Top Next Download PDF FiguresReferencesRelatedInformation Cited ByBlood Flow in Portal Systems with Special Reference to the Rat Pituitary Gland28 June 2016 | Journal of Cerebral Blood Flow & Metabolism, Vol. 12, No. 1Blood flow and functional responses correlate in the ovine neural lobeBrain Research, Vol. 373, No. 1-2Effects of sphere size and injection site on regional cerebral blood flow measurements.Stroke, Vol. 14, No. 5Cholecystokinin releases β-endorphin from the anterior pituitary glandNeuropeptides, Vol. 2, No. 6Release of vasopressin from the microdissected median eminence of the ratBrain Research, Vol. 206, No. 2The Effects Trauma and DMSO on Rat Skin Nutritive Blood FlowJournal of Investigative Dermatology, Vol. 62, No. 1The nervous control of blood flow to the pineal bodyLife Sciences, Vol. 6, No. 19Dye permeability and alkaline phosphatase activity of testicular capillaries in the postnatal ratHistochemie, Vol. 9, No. 4 More from this issue > Volume 16Issue 4July 1961Pages 762-764 Copyright & PermissionsCopyright © 1961 the American Physiological Societyhttps://doi.org/10.1152/jappl.1961.16.4.762PubMed13706540History Published online 1 July 1961 Published in print 1 July 1961 Metrics

The Blood-Brain Barrier

ArticlesThe Blood-Brain BarrierJ. DobbingJ. DobbingPublished Online:01 Jan 1961https://doi.org/10.1152/physrev.1961.41.1.130MoreSectionsPDF (10 MB)Download PDF ToolsExport citationAdd to favoritesGet permissionsTrack citations ShareShare onFacebookTwitterLinkedInWeChat Previous Back to Top Next Download PDF FiguresReferencesRelatedInformation Cited ByA definitive guide to the blood-brain barrierWillis K. 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