Urinary bladder function 6 months after the onset of diabetes in the spontaneously diabetic BB rat.

Urinary bladder function was examined in the spontaneously diabetic BB rat six months after the onset of diabetes. Diabetes caused significant decreases in rat weight and increases in bladder body weights and in vivo bladder capacities compared to age-matched controls, but no changes in the weights of bladder bases. The absolute contractile responses of urinary bladder body and base strips to nerve stimulation, carbachol, 5-hydroxytryptamine, ATP, phenylephrine, and KCl were unaltered by diabetes. However, when the data were corrected for tissue mass, there were slight but not significant decreases in contractile responses of strips from diabetic rats. There were increases in total muscarinic receptor numbers and calcium channel binding sites in bladder bodies from BB rats as a result of the increases in tissue mass. The data indicate that the six month-diabetic BB rat differs from the streptozotocin-diabetic rat in the sensitivity of the urinary bladder to the complications of diabetes, probably as a result of the insulin treatment required to keep BB rats alive.     

The evaluation of sympathetic system-related contractile activity of the rat vas deferens after ligation and intra-abdominal placement of the testis.

OBJECTIVE: To evaluate the contractile response of the vas deferens in a model of stress, to determine any changes in sympathetic activity as a result of stress in the ipsilateral testis, which decreases blood flow to the contralateral testis. MATERIALS AND METHODS: The study comprised two groups of six rats each; group 1 underwent a sham operation, and in group 2 the right testis was placed into the abdominal cavity and the vas deferens ligated. After 30 days, the vasa deferentia were resected bilaterally and their isometric contractions recorded. Electrical-field stimulation (EFS) was applied through a pair of platinum electrodes and concentration-response curves constructed for noradrenaline at 37 degrees C and to a solution containing 80 mmol/L K+. RESULTS: The vasa deferentia in both groups showed similar contractile responses to EFS, which were frequency-dependent and maximal at 80 Hz. Noradrenaline-induced contractile activity was lower in amplitude in the vasa deferentia of group 2 than in the contralateral and ipsilateral vasa deferentia of group 1, which were not significantly different from each other. All groups responded similarly to high K+. CONCLUSION: Intra-abdominal placement of the testes with vas deferens ligation decreased the contractile response to noradrenaline in the ipsilateral vas deferens without altering the contractile response to EFS and high K+. This difference could be caused by a reduction in the number of postjunctional alpha-adrenergic receptors or decreased receptor sensitivity. Both possibilities suggest that the vas deferens may initiate sympathetic activity, which may be responsible for contralateral testicular deterioration.     

Effect of thyroxine on the contractile responses of the vas deferens to prostaglandin E2

The effects of prostaglandin E2 (PGE2) on the contractile activity of thyroidectomized and thyroxine-treated albino rats were studied in vitro. Thyroidectomy totally inhibited the contractile response of the vas deferens to PGE2. Thyroxine treatment, on the other hand, significantly (P < .001) potentiated the response of vasa deferentia to PGE2, when compared to controls. It is suggested that thyroid hormones play a role in the contractile response of the vas deferens to PGE2.     

Changes in bladder function in the one year spontaneously diabetic BB rat.

Micturition characteristics and in vitro urinary bladder function were investigated in insulin-treated spontaneously diabetic BB rats and age-matched non-diabetic controls one year after the onset of diabetes. BB rats weighed less than controls and were hyperglycemic. Diabetic rats consumed larger volumes of water and excreted larger volumes of urine than controls. The frequency of micturitions and the mean volumes of urine excreted per micturition were significantly increased in BB rats compared to age-matched controls. Associated with the micturition changes in the BB rats were significant increases in bladder body mass. Contractile responses of strips from bladder bodies and bases were measured in response to nerve stimulation, carbachol, phenylephrine, ATP, and KCl. No significant differences between controls and diabetics were found in the absolute contractile responses of bladder body strips to nerve stimulation, carbachol, ATP, or KCl. However, if the data were transformed to correct for the increases in tissue mass in the diabetics, there were significant decreases in the responses of bladder body strips from BB rats to carbachol, ATP, and KCl, but not to nerve stimulation. Even after transformation, there were no differences in the responses of bladder base strips to carbachol, phenylephrine, or KCl. The data indicate that significant changes in micturition characteristics are evident one year after the onset of diabetes in the spontaneously diabetic BB rat. These changes are slow in development, since they are absent six months after the onset of diabetes. The changes in micturition and bladder strip contractility are qualitatively similar to, but quantitatively modest in comparison with those caused by streptozotocin-induced diabetes mellitus. The quantitative differences are probably attributable to an ameliorative effect of the insulin received by the BB rat.     

Early microrecanalization of vas deferens following biodegradable graft implantation in bilaterally vasectomized rats

We evaluated a biodegradable graft for reconstruction of rat vasa deferentia with long obstructed or missing segments. A total of 47 Sprague-Dawley rats underwent bilateral vasectomy and were divided into groups according to length of the vas deferens affected （0.5, 1, 1.5 cm）. After 8 weeks, poly-（D,L-lactide） （PDLA） grafts were used to reconnect the vas deferens, Grafts and adjoining vasa deferentia were excised 8 and 12 weeks later and evaluated microscopically. At 8 weeks, microscopic changes included a robust inflammatory response around the grafts. All grafts were still intact but in the early stages of degradation. No microtubules, indicative of vas deferens recanalization, were identified. One specimen showed evidence of healing and neovascularization at the interface zone between the vas deferens and the graft. At 12 weeks, grafts were further degraded but still present. Microscopic evaluation showed decreased inflammation. Seven specimens showed neovascularization at the interface zone; two of these showed distinct epithelialized vas deferens microcanals at the graft edges. One specimen showed a microcanal spanning the entire 0.5-cm graft. A time period of 8 weeks is not ample enough for vas deferens regeneration in the setting of a biodegradable PDLA graft; however, early evidence of re-growth was seen at 12 weeks. A longer healing time should permit further biodegradation of the graft, as well as re-growth and possible eventual reconnection of the vas deferens, allowing passage of sperm. These findings suggest a potential role for biodegradable grafts in the reconstruction ofvas deferens with long obstructed segments.     

Differential expression and regulation of integral membrane protein 2b in rat male reproductive tissues

Aim： To examine the expression and regulation of integral membrane protein 2b （Itm2b） in rat male reproductive tissues during sexual maturation and under different treatments by in situ hybridization. Methods： Testis, epididymis, and vas deferens were collected on days 1-70 to examine Itm2b expression during sexual maturation. To further examine the regulation of Itm2b, adult rats underwent surgical castration and cryptorchidism. Ethylene dimethane sulfonate and busulfan treatments were carried out to test the regulation of Itm2b after destruction of Leydig cells and germ cells. Results： In testis, Itm2b expression was moderately detected in the adluminal area of seminiferous cords on days 1-10, and detected at a low level in the spermatogonia on days 20 and 30. The Itm2b level was markedly increased in Leydig cells from day 20 to day 70. In epididymis and vas deferens, Itm2b was detected from neonate to adults, and the signal gradually increased in accordance with sexual maturation. Itm2b expression was significantly downregulated in epididymis and vas deferens of castrated rats, and strongly stimulated when castrated rats were treated with testosterone. Cryptorchidism led to a significant decline of Itm2b expression in testis and caput epididymis. Itm2b expression in epididymis and vas deferens was significantly decreased after the Leydig ceils were destroyed by ethylene dimethane sulfonate. Busulfan treatment produced no obvious change in Itm2b expression in epididymis or vas deferens. Conelusion： Our data suggested that Itm2b expression is upregulated by testosterone and might play a role in rat male reproduction.     

Effects of mexiletine on electrical field stimulation-induced contractile responses in the ipsilateral and contralateral vasa deferentia after unilateral testicular torsion/detorsion

AIM: To investigate testicular torsion-induced changes on the electrical field stimulation (EFS)-induced contractions in rabbit vasa deferentia and to evaluate the effect of mexiletine. METHODS: 18 male New Zealand albino rabbits were used in this experiment. Rabbits were divided into three groups: (1) control group (n = 6); (2) torsion group (n = 6), and (3) mexiletine group (n = 6). In the control group, vasa deferentia on both sides were harvested. In the torsion and mexiletine groups, the left testes of the rabbits were subjected to 720 degrees of clockwise torsion for 2 h and then detorsion was performed. In the mexiletine group, 50 mg/kg i.p. mexiletine was administered 1 h before detorsion. Following 24 h of the torsion, vasa deferentia on both sides were harvested and 2-cm strips including both the prostatic and epididymal portions were prepared to record EFS-induced contractions. RESULTS: Testicular torsion caused a significant inhibition in both phases of EFS-induced biphasic contractions of the ipsi- and contralateral vasa deferentia. Mexiletine treatment did not affect these inhibitory responses. Torsion/detorsion of the spermatic cord did not alter exogenously applied noradrenaline-induced contractions in both vasa deferentia. However, KCl-induced contractions diminished significantly in ipsilateral vas deferens of the torsion group and mexiletine restored this inhibition. CONCLUSIONS: Unilateral testicular torsion/detorsion leads to inhibition in both phases of EFS-induced biphasic contractions of the ipsi- and contralateral vasa deferentia by causing a defect in presynaptic nerve transmission. However, mexiletine has no effect on this inhibition. Inhibition of the KCl-induced contractions in the ipsilateral vas deferens, which indicates postsynaptic tissue damage, is restored by administering mexiletine 1 h prior to detorsion.     

Long-term use of sertraline leads to alterations in contractility of rat isolated vas deferens

Previous experiments with rat isolated vas deferens have shown that sertraline pretreatment inhibits contractile responses to noradrenaline, KCl, serotonin and electrical field stimulation. In the present study, the aim was to investigate the effects of long-term use of sertraline on contractile responses of rat isolated vas deferens. Fifteen Sprague-Dawley rats were given long-term (21 days) sertraline treatment, while another 15 were used as control. Both vas deferens were excised. Epididymal and prostatic segments of each underwent electrical field and chemical stimulation (noradrenaline, serotonin, acetylcholine, adenosine-triphosphate). Epididymal and prostatic segments had different contraction characteristics. Long-term sertraline treatment inhibited contractile responses of vas deferens segments to electrical field stimulation. The responses to noradrenaline were amplified with a left shift on both segments. Responses to serotonin had only a left shift on epididymal segments, while no contractile responses were observed on prostatic segments of the groups. Long-term treatment with sertraline had peripheral effects on rat vas deferens contractility, and some of the effects may be through mechanisms other than the inhibition of serotonin re-uptake.     

Biochemical and functional characteristics of bladder muscarinic receptors and effects of experimental diabetes in rats]

Bladder dysfunction is a common complication of diabetes mellitus and is attributed in part to peripheral neuropathy. Voiding function is mainly controlled by muscarinic receptor function. Therefore, I investigated first the biochemical and functional characteristics of urinary bladder muscarinic receptors and then the effects of experimental diabetes on them. Experimental diabetes was induced in 2 month-old male rats by intravenous injection of 65 mg/kg of streptozotocin (STZ). Effects of diabetes mellitus were investigated 2, 4 and 8 weeks after injection of STZ. The amount of muscarinic receptors labelled with 3H-quinuclidinyl benzylate (QNB) was higher in the bladder dome of diabetic animals than control animals, while the affinity for its binding sites was similar in both groups. Muscarinic agonists and antagonists inhibited 3H-QNB binding with similar inhibitory constants (Ki) in control and diabetic domes. The rank order of inhibition of 3H-QNB binding by muscarinic agonists and antagonists: bethanechol greater than pirenzepine greater than carbamylcholine greater than acetylcholine greater than atropine, is consistent with the absence of M1 receptors in the bladder dome. In functional studies muscarinic agonists induced a larger contractile response in bladder dome muscle strips from 8 week-old diabetic animal than those from controls. The rank order of ED50s were similar in the control and treated groups, being in good agreement with the Ki values obtained from receptor binding studies. These data show a direct correlation between the diabetes-induced biochemical and functional alterations in muscarinic receptor properties of the rat bladder.     

Effect of extracellular Mg concentration on electrically induced contractions of rat vas deferens in vitro

Contraction of smooth muscles of the vas deferens plays an important role in the propulsion of sperm into the pelvic urethra. This study examined the influence of external Mg2+ concentration on reactivity of the rat vas deferens to electrical stimulation in vitro. Vasa deferentia isolated from adult male rats were set up in tissue baths containing physiological salt solution at 37 degrees C and were stimulated electrically. Thereafter, increasing concentrations of Mg2+ were added to the bath and their effects on electrically evoked contractions were recorded. The effect of external Mg2+ depletion on evoked contractions was also examined. External Mg2+ depletion enhanced the contractile response to electrical stimulation while increasing external Mg2+ concentration inhibited the contractions. The inhibitory effect of Mg2+ was partially reversed by increasing extracellular Ca2+ concentration and was not additive with nifedipine. The results indicate that reactivity of the vas deferens to electrical stimulation is modulated by extracellular Mg2+ concentration. The possible relevance of these data to sperm transport through the vas deferens is discussed.     

Treatment of vas deferens large defects

Based on the experience of 12 vas deferens reconstructions in 8 patients with iatrogenic (post-herniotomy in infancy) vasa lesions the authors conclude that in the majority of such cases the length of vas defects renders direct vasovasoanastomosis either impossible or too risky, due to tension. To solve the problem the methods of extraanatomical (sub- and suprapubic) vas rerouting were elaborated that allow to shorten the vas length necessary for anastomosing by 9–14 cm. Combined with various crossover techniques, and vasoepididymostomy if necessary, the method proved valuable in most cases of extensive mono- and bilateral vasa deferentia lesions. The experimental data on vas segment transplantation obtained in rats show poor results in the homotransplantation group while autotransplants survive in 70% and provide vas patency restoration in 40%. No experimental proof in favour of endoprosthesing of vas grafts is obtained.     

The effect of biological dyes and contrast media on the vas deferens in Long Evans rats

The conventional diagnostic procedure of vasography utilizes a contrast medium to evaluate the patency of the vas deferens. With the development of microsurgical reconstruction for obstructive azoospermia in the past two decades, intraoperative vasography with saline or biological dye injection has replaced the use of radiographic contrast media. However, there are few reports on the effect of biological dyes on the healthy vas deferens. Therefore, we used experimental vasography to evaluate histological changes and functional patency of the vas deferens after infusion with a contrast medium and biological dye.
Four groups of 10 Long Evans male rats were injected by vasopuncture with 1% methylene blue, 1% gentian violet and 38% Urografin or saline into the vas deferens. The animals were killed 30 days later, and the vasa deferentia were excised and examined for histological changes and for functional patency. Vasopuncture with saline injection induced minimal change both at the puncture site and in the distal vas deferens. In both the Urografin- and methylene blue-injected groups, inflammation at the puncture site was found in 20–22% of cases, and 10–11% of cases revealed functional obstruction of the vasal lumen. In the gentian violet-injected group, severe histological and obliterated changes were found in all cases. Leakage of the dye and contrast medium or the sperm reaction may be responsible for the inflammation; otherwise, methylene blue and urografin did not seem to be harmful to the vas deferens. Although gentian violet is a blue dye, as is methylene blue, it has marked destructive effects on the vas deferens. It is concluded that some biological dyes used for vasal injection can cause occlusion of the vasal lumen, while inflammatory responses can occur from placing a needle transmurally.     

Identification and androgen-dependence of proteins in the mouse vas deferens

Proteins from 'luminal fluid' or from a homogenate of whole vas deferens were analysed by polyacrylamide gel electrophoresis under denaturing conditions. Four major bands with apparent molecular weights of 34.5, 36, 38 and 180 Kilodaltons (K) were observed in homogenates. In 'luminal fluid' the same protein pattern was observed except that 38 K band was missing. These four major bands probably originated from the vas deferens as they were not detected in plasma and were still present after ligation between the epididymis and vas deferens. After castration, there was a specific reduction of the 34.5 K MW protein band in both homogenate and 'luminal fluid'. When the androgen-dependence of proteins was investigated using radioactive methionine, the protein spectra from normal and castrated males showed that about 10 polypeptides were differentially induced or repressed by androgens. The synthesis of some proteins (MW's 24, 36 and 180 K) was decreased by castration while the synthesis of several others (MW's between 27 and 82 K) was increased. Most of these androgen-dependent proteins were detectable at 10 days of age.     

Robotic assisted microsurgical vasal reconstruction in a model system

Our objective was to determine whether or not male reproductive microsurgery is adaptable to current robotic technology. We devised a model vas deferens system using rat vasa deferentia and completed vasal anastomosis with full-thickness and mucosal robotically placed, Sharpoint 10-0 bicurve nylon sutures. Experienced and inexperienced microsurgeons performed separate anastomoses. Both groups of microsurgeons completed anastomoses with accuracy and enhanced comfort. The robotic graspers had the dexterity to delicately handle the 10-0 sutures and needles. We conclude that through our model vas deferens system, we demonstrated the feasibility of applying robotic technology to male reproductive microsurgery. This pilot study reveals some advantages of the robot and suggests future use of this system in a new urological application domain.     

The function of autonomic receptors in canine ureteral smooth muscle]

I have investigated the effects of autonomic drugs and prostaglandins on in vitro smooth muscle spontaneous contractions and made the quantitative analysis of autonomic receptors in the canine ureter. Ureteral muscle strip cut helically usually generated spontaneous contractions whereas those cut circularly or longitudinally did not generate spontaneous contractions. These results suggest the importance of knowing which direction to cut the ureteral smooth muscle in order to generate spontaneous contractions. Norepinephrine (alpha), phenylephrine (alpha 1), carbachol (muscarinic) and PGF2 alpha caused significant increase in contractile force. Terbutaline (beta 2) and PGE2 caused significant decreases in contractile force, while dobutamine (beta 1) and clonidine (alpha) caused no effect. Autonomic receptor densities were determined using radiology and binding techniques. The number of maximum binding sites (Bmax) of 3H-prazosin (PZ), 3H-yohimbine (YOH), 3H-dihydroaloprenolol (DHA) and 3H-quinuclidinylbenzilate (QNB) were 53.8, 16.9, 11.2 and 5.28 fmol/ml protein, respectively. These data suggest that the contractile responses to adrenergic and cholinergic agonists in the canine ureter are mediated through functional adrenergic (alpha 1, beta 2) and muscarinic cholinergic receptors and that the prostaglandins have a role in the contraction of the canine ureter.     

Duplicated vas deferens: A case report and comprehensive review of the literature

Duplication of vas deferens is a very rare anomaly which two vasa deferentia are found in the spermatic cord. It can be recognised during autopsy or cadaveric dissection and also several surgical procedures which require spermatic cord dissection including inguinal hernia repair, orchiopexy, vasectomy, varicocelectomy, vasectomy reversal and radical prostatectomy. Recognition of the duplicated vas deferens is important to avoid surgical complications such as an unsuccessful vasectomy or transection of the vas. It was reported in only three cadavers and 31 patients since 1959. In this study, we describe a new case of duplicated vas deferens found incidentally during routine inguinal hernia repair in a 66-year-old patient. We also review all previously reported cases in the literature to draw attention to this rare but important anomaly.     

Histological study of vas deferens following intravasal laser irradiation

Aim: To study the histologic changes of the vas deferens following Nd: YAG laser irradiation. Methods:Intravasal laser irradiation was given to (i) 52 segments of rabbit (laser dosage: 2 seconds at 40W～50W) and 16 segments of human (3 seconds at 45W～55W) vas deferens in vitro, (ii) 25 rabbit vasa (2 seconds～2.5 seconds at 40W～45W) in vivo and (iii) 2 human vasa (3 seconds at 55W) in vivo. Segments of vasa were removed from the in vivo irradiated vasa deferentia 15 days ～ 180 days (rabbit) or 15 days (man) after the exposure. All vas segments were embedded in methacrylate resin. Serial sections (thickness 25μm～30μm) were obtained and observed under a light microscope. Results: (i) Laser-induced damage reached the muscularis layer in 27% and 94% of the rabbit and human vas segments in vitro, respectively. (ii) Fourteen of the 25 in vivo rabbit vasa were completely occluded by fibrous tissue and the longer the time interval after treatment, the more likely was the vas occluded. Those unoccluded vasa had either a normal histology or a mucosal damage. (iii) One in vivo human vas was almost completely occluded by the fibrous tissue but the other had a relatively large lumen packed with sperm granulomatous tissue and partial destruction of the smooth muscle layer. Conclusion: Laser irradiation can induce long-term vas occlusion; for rapid occlusion, laser doses just completely destroying the mucosal layer will be advisable.     

Crossed ectopic testis with common vas deferens

Crossed ectopic testis is an uncommon anomaly. A 10-month-old boy with a crossed ectopic testis and a common vas deferens is presented. At operation 2 testes were in the left inguinal canal. The seminal vesiculogram showed that both vasa deferentia fused to form a common lumen, and then formed a common stem on the left side and opened into the posterior urethra. The spermatic arteriogram revealed that each testis was supplied from the right and left sides, respectively. No testis or vas deferens was located in the right retroperitoneal space. These features suggested that the testes developed from both sides and subsequently descended to the left side.     

Spontaneous occurrence of vasculitis-like lesions in male reproductive tissues in mice: a histological study.

Vasculitis involving testes, epididymides, and vasa deferentia is considered to be a rare disease but has often been reported in man. In the present study, reproductive tissues of young and aged mice were examined to determine whether spontaneous vasculitis occurs in them. Testes, ductuli efferentes, epididymides, and vasa deferentia were obtained from young and aged BALB/c mice together with some nonreproductive organs for histological observation at the light microscopic level. The examination revealed that vasculitis-like lesions comparable to those in man were significantly present in the epididymides and vasa deferentia of aged but not young mice. However, no significant lesions were found in the testes and ductuli efferentes. Furthermore, other organs, such as salivary glands, thyroid glands, livers, pancreases, and kidneys, were also negative for the lesions. These results indicate that the epididymis and vas deferens are spontaneously apt to be affected by vasculitis-like lesions with advancing age, but that the lesions are not due to a systemic vasculitis disease.