Vas Deferens Fluid Research Articles

Vasectomy re-reversal: effectiveness and parameters associated with its success.

ABSTRACTIntroduction:When the vasectomy reversal (VR) fails, and the patient desires natural conception with his sperm, vasectomy re-reversal (VRR) is the only alternative.Purpose:To determine the VRR effectiveness and whether specific parameters can be associated with its success.Materials and Methods:We retrospectively evaluated 18 consecutive vasectomized patients, who had failed their VR through bilateral vasovasostomy, and posteriorly were submitted to VRR. The parameters of the study were: age of the patients, elapsed time between vasectomy and VRR (V-VRRt), elapsed time between VR and VRR (VR-VRRt), presence of spermatozoa in the proximal vas deferens fluid (SptzVDF) in the VRR and results of semen analysis after VRR (SA-VRR).Results:The mean of the age of the patients was 44.11±6.55 years (32.0-57.0), the mean of V-VRRt was 11.76±6.46 years (1.5-25.0) and the mean of VR-VRRt was 2.13±2.27 years (0.5-10.0). SptzVDF in the VRR were found bilaterally in 8 patients, unilaterally in 4 and absent in 6. SA-VRR demonstrated normozoospermia in 9 patients, oligozoospermia in 3 and azoospermia in 6, with patency rate of 66.67%. SA-VRR showed statistically significant dependence only with SptzVDF in the VRR (p <0.01).Conclusions:VRR was effective in restoring the obstruction in more than half of the patients. Furthermore, the presence of spermatozoa in the vas deferens fluid was the parameter associated with the VRR success.

Localization of sperm intracellular Ca2+ keeps fertilizability in the newt vas deferens.

Sperm intracellular Ca2+ is crucial for the induction of sperm-egg interaction, but little is known about the significance of Ca2+ maintenance prior to induction. In sperm of the newt Cynops pyrrhogaster, intracellular Ca2+ is localized to the midpiece during storage in the vas deferens, while extracellular Ca2+ is influxed in modified Steinberg's salt solution to promote a spontaneous acrosome reaction related to the decline of sperm quality. In the present study, sperm from the vas deferens were loaded with the Ca2+ indicator Fluo8H, and changes in Ca2+ localization in modified Steinberg's salt solution were examined. Calcium ions expanded from the cytoplasmic area of the midpiece to the entire tail in most sperm during a 1-h incubation and localized to the principal piece in some sperm within 24 h. Similar changes in Ca2+ localization were observed in reconstructed vas deferens solution that included ions and pH at equivalent levels to those in the vas deferens fluid. Sperm with Ca2+ localization in the entire tail or the principal piece weakened or lost responsiveness to sperm motility-initiating substances, which trigger sperm motility for fertilization, but responded to a trigger for acrosome reaction. The change in Ca2+ localization was delayed and transiently reversed by ethylene glycol tetraacetic acid or a mixture of Ca2+ channel blockers including Ni2+ and diltiazem. These results suggest that C. pyrrhogaster sperm localize intracellular Ca2+ to the midpiece through Ca2+ transport in the vas deferens to allow for responses to sperm motility-initiating substances.

Hormonal control of vas deferens fluid volume and aquaporin expression in rats.

Precise regulation of vas deferens fluid volume which is important for sperm survival might be influenced by testosterone. In order to investigate changes in vas deferens fluid volume and aquoporins (AQP) isoforms expression under testosterone influence, orchidectomized Sprague-Dawley rats were given 125 and 250µg/kg/day testosterone with or without flutamide, an androgen receptor blocker or finasteride, a 5alpha-reductase inhibitor for seven consecutive days. Following treatment completion, vas deferens was perfused and changes in the fluid secretion rate and osmolality were determined in the presence of acetazolamide. Rats were then sacrificed and vas deferens was harvested for histology, tissue expression and distribution analyses of AQP-1, AQP-2, AQP-5, AQP-7 and AQP-9 proteins by Western blotting and immunohistochemistry, respectively. Our findings indicate that testosterone causes vas deferens fluid secretion rate to increase, which was antagonized by acetazolamide. Fluid osmolality increased following testosterone treatment and further increased when acetazolamide was given. Co-administration of flutamide or finasteride with testosterone causing both fluid secretion rate and osmolality to decrease. Histology revealed increased size of vas deferens lumen with increased thickness of vas deferens stroma. Expression of AQP-1, AQP-2 and AQP-9 were detected in vas deferens but not AQP-5 and AQP-7, and the levels of these proteins were increased by testosterone treatment mainly at the apical membrane of vas deferens epithelium. In conclusion, increased in vas deferens fluid secretion rate under testosterone influence mediated via the up-regulation of AQP-1, 2 and 9 might be important for vas deferens fluid homeostasis in order to ensure normal male fertility.

Testosterone enhances expression and functional activity of epithelial sodium channel (ENaC), cystic fibrosis transmembrane regulator (CFTR) and sodium hydrogen exchanger (NHE) in vas deferens of sex-steroid deficient male rats

Effects of testosterone on expression and functional activity of ENaC, CFTR and NHE in vas deferens were investigated. MethodsOrchidectomized, adult male rats were given 125 and 250 μg/kg/day testosterone subcutaneously, with or without flutamide and finasteride for seven consecutive days. At the end of the treatment, rats were anesthetized and vas deferens were perfused. Changes in vas deferens fluid secretion rate, pH, HCO3−, Cl− and Na+ concentrations were recorded in the presence of amiloride and Cftr inh-172. Rats were then sacrificed and vas deferens were harvested and subjected for molecular biological analysis. ResultsTestosterone treatment caused the fluid pH and HCO3− concentrations to decrease but secretion rate, Cl− and Na+ concentrations to increase, where upon amiloride administration, the pH and HCO3− concentration increased but Cl− and Na+ concentrations further increased. In testosterone-treated rats, administration of Cftr inh-172 caused all fluid parameters to decrease. In testosterone-treated rats co-administered with flutamide or finasteride, pH and HCO3− concentration increased but fluid secretion rate, Cl− and Na+ concentrations decreased and these parameters were not affected by amiloride or Cftr inh-172 administration. Under testosterone influence, CFTR and γ-ENaC were highly expressed at the apical membrane while NHE-1 and 4 were highly expressed at the basolateral membrane of vas deferens epithelium. Meanwhile, NHE-2 and 3 were highly expressed at the apical membrane. ConclusionsDifferential expression of ENaC, CFTR and NHE in vas deferens under testosterone influence indicated the important role of these transporters in creating optimal fluid microenvironment that is essential for preserving male fertility.

556 Does percutaneous epididymal sperm aspiration actually cause epididymal obstruction?

To evaluate the sperm in vas deferens fluid (VDF) after percutaneous sperm aspiration of epididymis (PESA) in order to establish whether there was patency of epididymal ducts and what parameter could predict this occurrence. Ten vasectomized patients after failed ICSI/IVF procedures, using PESA (8 unilateral, 2 bilateral), underwent intraoperative sperm analysis of VDF during their microsurgical vasectomy reversals. The parameters of the study were the age of the patients, the elapsed time between vasectomy and VDF analysis, between PESA and VDF analysis, and the presence, concentration and motility of sperm in VDF. The mean age of the patients was 47.50 ± 5.48 years (40 - 59). The mean elapsed time between vasectomy and VDF sperm analysis was 17.50 ± 7.58 years (10 - 25). The mean elapsed time between PESA and VDF analysis was 24.60 ± 17.00 months (7 - 54). The VDF analysis showed sperm in 5 patients, with mean concentration of (33.04 ± 24.27) million sperm/ml and motile sperm in 3 patients (progressive: 10%, 5% and <1%, respectively). The mean elapsed time between vasectomy and VDF analysis showed a statistically significant difference between the 5 patients who had sperm in the VDF and 5 who did not have sperm in the VDF (13.80 ± 6.34 and 21.20 ± 7.40 years, respectively, p <0.05).

Testosterone up-regulates vacuolar ATPase expression and functional activities in vas deferens of orchidectomized rats

Precise regulation of vas deferens fluid pH is essential for sperm. However, the mechanisms underlying effect of testosterone on vas deferens fluid pH have never been identified, which could involve changes in expression and functional activity of vacoular (V)-ATPase. MethodsOrchidectomized, adult male Sprague-Dawley rats were treated subcutaneously with 125 μg/kg/day and 250 μg/kg/day testosterone with or without flutamide (androgen receptor blocker) and finasteride (5α-reductase inhibitor) for seven (7) days. Following treatment completion, in vivo perfusion of vas deferens lumen was performed and changes in fluid secretion rate, pH and HCO3− content were measured with and without bafilomycin, a V-ATPase inhibitor. Rats were then sacrificed and vas deferens were harvested and subjected for V-ATPase A1 and B1/2 protein expression and distribution analysis by western blotting and immunohistochemistry, respectively. ResultsIn sham-operated and testosterone-treated orchidectomized rats, higher fluid secretion rate, which was not antagonized by bafilomycin but lower HCO3− content and pH which were antagonized by bafilomycin were observed when compared to orchidectomized-only and orchidectomized, testosterone-treated rats receiving flutamide or finasteride, respectively. Bafilomycin had no effect on fluid secretion rate, HCO3− content and pH in orchidectomized and testosterone-treated orchidectomized rats receiving flutamide and finasteride. V-ATPase A1 and B1/2 proteins were expressed at high levels in vas deferens and were highly distributed at the apical membrane of luminal epithelium and in muscle layer of this organ, mainly in sham and testosterone-treated orchidectomized rats. ConclusionsV-ATPase is involved in acidification of vas deferens fluid under testosterone influence.

Contribution of different Ca2+ channels to the acrosome reaction-mediated initiation of sperm motility in the newt Cynops pyrrhogaster

Initiation of sperm motility in urodeles, which is induced by a sperm motility-initiating substance (SMIS) in the sequestered granules on the surface of egg jelly, is mediated by the acrosome reaction (AR), which is triggered by an AR-inducing substance (ARIS) on a sheet-like structure. Details of the unique process of the interaction between egg jelly and sperm in these species is still unclear. The current study showed the fine structure of egg jelly in the newt Cynops pyrrhogaster, a urodele species, revealing that its outer surface was covered by a sheet-like structure of approximately 0.29 μm in thickness. Granules of approximately 2 μm in diameter with small particles of approximately 54 nm were attached to its surface and distributed inhomogeneously just beneath the sheet-like structure. Emission spectrometry revealed that the Ca2+ concentration was maintained at a high level compared with that of the blood plasma and the vas deferens fluid, suggesting that egg jelly is a reliable source of Ca2+ for the sperm-egg interaction. Blockers of the T-type voltage-dependent Ca2+ channel (VDCC), but not the L-type VDCC, inhibited both AR and initiation of sperm motility. Conversely, Ni+, which affects the α1 H subunit of T-type VDCC, only inhibited the initiation of sperm motility. These data suggest that, in response to ARIS and SMIS, sequential gating of distinct Ca2+ channels occurs in the AR, followed by the initiation of sperm motility on the surface of the egg jelly in C. pyrrhogaster at fertilization.

Sperm analysis of the vas deferens fluid after a long interval of unilateral percutaneous epididymal sperm aspiration in vasectomized patients

Evaluation of the presence of spermatozoa in vas deferens fluid after a long interval of unilateral and homolateral percutaneous epididymal sperm aspiration (PESA) in vasectomized men. When found, the spermatozoa were evaluated including concentration and motility, in order to verify the patency of the epididymal tubules. Four patients, numbered in a progressive order, from one to four, with 38, 40, 48 and 51 years old and vasectomy interval of 10, 10, 25 and 11 years, respectively, whose wives did not get pregnant using intracytoplasmic sperm injection of sperm obtained by unilateral PESA and decided to try only natural conception, were submitted to intrasurgical sperm analysis of the vas deferens fluid (ISAVDF) during microsurgery for reconstruction of the seminal tract. Time interval between PESA and ISAVDF was 13.75 ± 11.12 months (x ± s) varying from 3 to 29 months. Homolateral ISAVDF and PESA showed the presence of spermatozoa. Patients 1, 2 and 4 had a high concentration of 10 x 106, 64 x 106 and 45 x 106 spermatozoa/ mL; the first two had motile sperms and patient 3 had no sperms. Three of four patients showed spermatozoa in the vas deferens fluid after a long interval of unilateral and homolateral PESA with high concentration, including motile forms. These findings support the concept that PESA may not result in late epipidymal tubule obstruction in vasectomized patients.

Designed Self Assembly of Nano-Liposomes in the Male Reproductive Tract for Model Drug Delivery to the Prostate

Using the compound Styrene maleic anhydride (SMA) as a model of a drug, a novel approach for sustained endogenously formed liposome encapsulated drug delivery to the prostate gland has been evolved. Intra vas deferens de- pot of a combination of high molecular weight SMA (SMAh, the drug) and low molecular weight SMA (SMAl) is formed by one time injection. The SMAh breaks down sperm membrane to provide from the sperms a continuous supply of phos- pholipids. The SMAl forms cleavage centers causing slow breakaway of nano SMAh fragments. Vas peristaltic pressures provide the mechanical forces bringing reactants together leading to encapsulation of nano particles of SMAh within phospholipids cover giving a continuous supply of nano liposomes. A dimethyl sulphoxide (DMSO) constituent of the de- pot leads to sulfur attachment to the liposome on account of which by a prostate tissue sulfur affinity mechanism the nano liposomes traverse the vas deferens- prostate barrier. The concept has been tested by implantation of the SMA in the rat vas deferens. Formation of nano liposomes in vivo; actual encapsulation of SMA within the liposome; the overall drug Encapsulation Efficiency; presence of liposomes in the vas deferens fluid and transfer to the prostate have been confirmed by Transmission Electron Microscope (TEM) examination and Fluorescence microscopy following Nile Red staining of vas fluid and prostate tissue

Acquisition of arylsulfatase A onto the mouse sperm surface during epididymal transit.

Arylsulfatase A (AS-A) is localized to the sperm surface and participates in sperm-zona pellucida binding. We investigated how AS-A, usually known as an acrosomal enzyme, trafficked to the sperm surface. Immunocytochemistry of the mouse testis confirmed the existence of AS-A in the acrosomal region of round and elongating spermatids. However, immunofluorescence and flow cytometry indicated the absence of AS-A on the surface of live testicular sperm. In contrast, positive AS-A staining was observed in the heads of live caudal epididymal and vas deferens sperm. The results suggested that acquisition of AS-A on the sperm surface occurred during epididymal transit. Immunocytochemistry of the epididymis revealed AS-A in narrow and apical cells in the initial segment and in clear cells in all epididymal regions. However, these epithelial cells are in the minority and are not involved in secretory activity. In the caudal epididymis and vas deferens, AS-A was also localized to principal cells, the major epithelial cells. Because principal cells have secretory activity, they may secrete AS-A into the epididymal fluid. This hypothesis was supported by our results revealing the presence of AS-A in the epididymal and vas deferens fluid (determined by immunoblotting and ELISA) and an AS-A transcript in the epididymis (by reverse transcription polymerase chain reaction). Alexa-430 AS-A bound to epididymal sperm with high affinity (Kd = 46 nM). This binding was inhibited by treatment of sperm with an antibody against sperm surface sulfogalactosylglycerolipid. This finding suggests that AS-A in the epididymal fluid may deposit onto sperm via its affinity to sulfogalactosylglycerolipid.

Vasectomy reversal.

Vasectomy reversal.

Platelet-activating factor acetylhydrolase in the male reproductive tract: origin and properties.

The properties and origin of platelet-activating factor acetylhydrolase- (PAF-AH)-like activity in the male reproductive tract were investigated. Seminal plasma (SP) and serum were obtained from normal donors and infertile patients while prostatic fluid (PF), seminal vesicle fluid (SVF) and vas deferens fluid were collected at autopsy. PAF-AH-like activity was found in all fluids tested. The specific activity of the enzyme in SP and PF was twice that of PAF-AH in serum and 15-fold higher than that in SVF and vas deferens fluid. In SP, PAF-AH-like activity was Ca(++)-independent, acid and heat labile, stable to freezing, not inhibited by phosphatidylcholine, but was inhibited by 10 mM disopropylfluorophosphate (DFP) and 13 mM phenylmethylsulfonylfluoride (PMSF). These data indicate that the properties of the enzyme in SP are similar to those reported for PAF-AH in serum. The variation in specific activity of PAF-AH in reproductive tract fluids suggest that there are either activators of PAF-AH in SP or inhibitors in one or several of the other reproductive tract fluids.

An intravasal non-occlusive contraceptive device in rats

An implantable miniature biogalvanic cell was developed to kill the spermatozoa in the vas deferens by an electric current. Experiments in vitro and in vivo showed that a combination of aluminum/silver electrodes connected with platinum wires with the vas deferens fluid as n electrolyte is effective in killing spermatozoa. Female rats paired with males having biogalvanic devices in the vas deferens, which remained patent, did not produce any young.

Peristaltic pumping in non-uniform tubes

Peristaltic flow in non-uniform channels and tubes has been investigated with particular reference to the flow of spermatic fluid in vasdeferens. The pressure rise over the tube length at any specified flow rate and amplitude ratio is found to be much less compared to that in uniform geometry. A comparison, with the experimental data in vasdeferens in rhesus monkeys, shows that perstaltic mechanism can account for approximately one-third of the measured flow rate.