Early testicular changes after vasectomy and vasovasostomy in Lewis rats

The testes of Lewis rats were studied at intervals from 2 weeks to 3 months after bilateral vasectomy, vasectomy followed 1 month later by vasovasostomy, or sham operations. Aims were to determine the nature of early alterations after vasectomy, and to determine whether vasovasostomy after 1 month would result in reversal of vasectomy-induced changes. Approximately one-fourth of the testes in the vasectomy and vasovasostomy groups displayed histological changes, which consisted mainly of depletion of germ cells. The extent of the depletion varied greatly in different seminiferous tubules. In testes altered in this way, no abnormal infiltrations of lymphocytes, macrophages, or other cells were observed in the seminiferous epithelium or in the interstitium. The rete testis and straight tubules were normal in testes with altered seminiferous epithelium. A few testes in the vasectomy and vasovasostomy groups had necrotic centers. The results suggest that depletion of germ cells occurred as a result of shedding from the seminiferous epithelium into the lumen of the tubules. A cellular immune response, such as occurs in experimental allergic orchitis in other species, did not appear to be responsible for the observed loss of germ cells. This suggests a possible role for humoral antibody in this model, since there is an association between testicular changes and serum antisperm antibodies at longer intervals after vasectomy. Testicular alterations were not reversed by performance of a vasovasostomy 1 month after vasectomy.     

输精管结扎与吻合术后对兔睾丸细胞功能的影响

家兔31只,分为假手术对照组(C),输精管结扎组(V)和输精管吻合组,后者于结扎后12个月时行吻合术,吻合后3个月与雌兔配对交配,继续观察2个月,根据雌兔孕否再将吻合兔分为吻合育组(VaF)和吻合不育组(VaI)。结果表明,血清睾酮水平在C,VaF,VaI和V组分别是7.1±4.2,11.3±4.7,4.3±2.3和4.9±2.7nmol/L((?)±SD,下同),VaF,VaI和V组分别与C组比较差异不显差(P>0.05),而VaF和VaI组比较差异显著(P<0.01),睾丸细胞核雄激素受体浓度在C、VaF,VaH和V组分别是61.1±17.8,66.2±38.2,44.5±26.8和68.9±22.5fmol/mg.DNA,组间比较无显著差异(P>0.05);睾丸组织cAMP含量在C,VaF,VaI和V组分别是440.0±94.0,324.0±13.0,277.0±48.0和254.0±135.0pmol/g组织,VaF,VaI和V组显著低于C组(P<0.05);而睾丸胞液雄激素结合蛋白浓度分别是27.4±10.5,64.8±18.5,52.8±23.5和44.7±14.7fmol/mg·pro.,VaF,VaI和V组显著高于C组(P<0.05),睾丸细胞膜Na~ ,K~ -ATPase活性分别是78.0±28.7,62.2±10.3,43.7±9.8和32.6±10.0μmol Pi/mg·hr,VaI和V组显著低于C和VaF组(P<0.01),而Mg~(2 )-ATPase活性分别是57.2±27.0,53.6±16.2,30.9±10.9和23.1±9.1μmol Pi/mg.hr,VaI和V组显著低于VaF和C组(P<0.05,P<0.01)。     

Ultrastructure of epididymal interstitial reactions following vasectomy and vasovasostomy

The response of the male reproductive tract to vasectomy includes inflammation of the interstitial tissue of the epididymis. The pathogenesis of epididymal interstitial reactions and characteristics of the responding cells were studied by electron microscopy in Lewis rats at intervals following bilateral vasectomy, vasectomy followed 1 month later by vasovasostomy, or sham operations. In areas of interstitial reaction, numerous macrophages, monocytes, lymphocytes, neutrophils, and plasma cells occupied the connective tissue. Macrophages, containing many lysosomes and vesicles, aggregated and assumed the appearance of epithelioid cells. Processes of adjacent macrophages interdigitated with one another and closely approached the surfaces of lymphocytes. Many plasma cells with distended rough endoplasmic reticulum appeared in the interstitium. The majority of animals in the vasectomy and vasovasostomy groups exhibited epididymal interstitial changes by 2–3 months; the cauda epididymidis was the region most often affected. The ultrastructural features were indicative of chronic granulomatous inflammation and were consistent with an immune response that includes antigen presentation by macrophages to lymphocytes, lymphocyte differentiation, and local antibody production by plasma cells. The nearly complete absence of sperm or recoghizable parts thereof in the interstitial tissue in the areas of the reactions suggests that these lesions formed in response to soluble antigens leaking from the duct. Vasovasostomy was not effective in reversing or retarding epididymal inflammation at the intervals studied. © 1993 Wiley-Liss, Inc.     

输精管吻合术后不育原因的实验研究精子密度与睾丸功能

14只成年日本大耳白兔,双侧输精管结扎12月后,以显微外科技术行双侧输精管吻合术,3月后与雌兔配对交配,观察2个月,根据妊娠与否分为输精管吻合育组(VFG)和输精管吻合不育组(VIG),各7只。另设输精管结扎组(VG)和假手术组(SOG)作为对照。结果表明,(1)VFG的精子密度与SOG比较虽为低值(P<0.01),但显著地高于VIG(P<0.01)。(2)精子密度与睾丸ACE活力、Na~+,K~+-ATPasc活力、Mg~(++)-ATPasc活力、睾丸cAMP含量均呈显著的正相关。(3)精子密度、cAMP含量与ABP呈明显的负相关。(4)VFG血清睾酮含量与VIG比较有显著差异(P<0.05)。VFG、VIG中血清睾酮水平与精子密度呈明显的正相关(r=0.60、P<0.05)。     

Morphological alterations and distribution of occludin in rat testes after bilateral vasectomy

The aim of study was to investigate the fate and the morphology of the cells which constitute the spermatogenic line, and to determine the distribution of occludin in the testis in adult vasectomized Wistar rats. The rats were divided into two groups: control group (sham-operated) and vasectomized group. One, 3 and 6 months after sham and vasectomy operations, testis samples were examined. The weight of the testes was found to be reduced 3 and 6 months after vasectomy. There was vacuolization in the seminiferous tubules one month after vasectomy. The tubules showed severe atrophy 3 and 6 months after vasectomy. The occludin immunolabeling in the 3- and 6-month groups was weak and diffuse, and the density of the protein was found to be decreased. The increase in the number of apoptotic cells was accompanied by a time-dependent decrease in the number of haploid, diploid and tetraploid cells. This study demonstrated that vasectomy causes degeneration in the seminiferous tubules with alterations in occludin distribution with a decrease in the number of spermatogenic cells. Moreover, these alterations increase in a time-dependent manner.     

Effects of vasectomy on spermatogenesis and fertility outcome after testicular sperm extraction combined with ICSI

BACKGROUND: Each year 40,000 men have a vasectomy in the UK whilst another 2400 request a reversal to begin a second family. Sperm can now be obtained by testicular biopsy and subsequently used in assisted conception with ICSI. The study aims were to compare sperm yields of men post-vasectomy or with obstructive azoospermia (OA) of unknown aetiology with yields of fertile men and to assess any alteration in the clinical pregnancy rates after ICSI. METHODS: Testicular tissue was obtained by Trucut needle from men who had undergone a vasectomy >5 years previously or had OA from other causes and from fertile men during vasectomy. Seminiferous tubules were milked to measure sperm yields. Numbers of Sertoli cells and spermatids and thickness of the seminiferous tubule walls were assessed using quantitative computerized analysis. RESULTS and CONCLUSIONS: Sperm yields/g testis were significantly decreased in men post-vasectomy and in men with OA, relative to fertile men. Significant reductions were also observed in early (40%) and mature (29%) spermatid numbers and an increase of 31% was seen in the seminiferous tubule wall (basal membrane and collagen thickness) of vasectomized men compared with fertile men. Clinical pregnancy rates in couples who had had a vasectomy were also significantly reduced.     

Testicular protective effects of ellagic acid on monosodium glutamate-induced testicular structural alterations in male rats

ABSTRACT

Ellagic acid (EA) has significant protective and antioxidant effects on several tissues. Monosodium glutamate (MG) is known as a flavor promoter that reversibly influences the male reproductive system. This study aims to assess the ameliorative effect of EA on oxidative stress and testicular damage induced by MG. In total, 48 male rats were included in this study and separated into six groups: control, EA (20 mg/kg), MG (low dose) (17.5 mg/kg), MG (high dose) (60 mg/kg), MG (low dose) combined with EA, and MG (high dose) combined with EA. Testicular antioxidant biomarkers [superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GRx), catalase (CAT), myeloperoxidase (MPO), and xanthine oxidase (XO)] were examined. Testes were examined and scored for histological variation as an indicator of testicular damage following administration of MG alone or in combination with EA. Serum testosterone, inhibin B, 8-hydroxydeoxyguanosine (as a marker of DNA damage), and transmission electron microscope sections of the testis were evaluated, and a comet assay was performed. Results showed that administration of EA combined with MG significantly elevated the levels of enzymatic antioxidants and decreased lipid peroxidation compared with MG treatment alone. EA elevated testosterone hormone levels and thus enhanced male reproductive capacity. It is clear from the data that EA inhibits histological and ultrastructure testicular damage and improves the redox state in male rats.     

Pregnancy after vasovasostomy for vasectomy reversal: a study of factors affecting long-term return of fertility in 282 patients followed for 10 years

The aim of this study was to determine the eventual fertilityof those patients following vasectomy reversal who have no pressure-inducedsecondary epididymal blockage. These patients underwent simplevasovasostomy because at the time of the reversla surgery therewere sperm present in large numbers in the vas fluid. It waspossible to obtain long-term follow-up on 326 early patientswho underwent vasectomy reversal 8–10 years age. Two hundredand eighty-two of those patients had sperm in the vas fluid.These patients, were studied for pregnancy rate and post-operativesemen parameters in relation tio presence or absence of spermin the vas fluid at the time of vasectomy reversal, durationof time since vasectomy, pre-operative serum antisperm antibodytiters, the influence of varicocoele and quantitative evaluationof testicular biopsy. All of the 44 patients with no sperm inthe vas fluid remained azoospermic following vasovasostomy.Of the 282 patients with sperm in the vas fluid, 228 (81% )eventually impregnated their wives. Twenty-four patients withsperm in the vas fluid (9%) were azoospermic and did not impregnatetheir wives. Of the 258 patients who had sperm patency, thepregnancy rate was 88%. The number of mature spermatids pertubule in the testis correlated closely with the post-operativesperm count in patent cases. Quantitative evaluation of thetesticular biopsy revealed normal spermtogenesis, even in patientswith azoospermia or severe oligospermia post-operatively. Technicalfailures were due to blockage either at the vasovasostomy site,or epididymal blockage unrecognized at the time of vasovasostomy.Sperm count had a minimal impact on the likelihood of pregnancyso long as there was patency, and there was no discrepancy betweensperm count and actual testicular sperm production as determinedby testicular biopsy. Pregancy was not related to the presenceor absence of a varicocoele, pre-oprative serum sperm antibodylevels, or testicular biopsy findings     

The fate of sperm after vasectomy in the hamster

Young adult male hamsters were subjected to bilateral vasectomy. The reproductive tracts were studied by light and electron microscopy at intervals up to 1 year after the operation. Sperm continued to be produced, since testicular alterations were focal. Spermatic granulomas were associated with the excurrent ducts of all animals 5 months or 1 year after vasectomy and with those of one of four hamsters 2 weeks after the operation. Phagocytosis of sperm in the lumina of the efferent ducts and proximal parts of the epididymis, and disintegration of membranous components of intraluminal sperm occurred in approximately three-fourths of the animals studied 5 months or more after vasectomy. The results indicate that after vasectomy in the hamster sperm are disposed of by phagocytosis in spermatic granulomas, intraluminal phagocytosis, and dissolution in the lumen of the male ducts, although the latter process may be incomplete.     

Changes in seminiferous tubules after vasectomy

The histologic, immunohistochemical, and ultrastructural changes in the seminiferous tubules (ST) of 17 healthy adult males who had been vasectomized between 1 1/2 and 5 years are reported. There was minimal spermatogenesis in 4 cases. 8 of the cases underwent electron microscopy examination and 4 showed evidence of minimal histological spermatogenesis. The ST showed a thickening of the basement membrane and heavy deposits of lipofuchsin in the Sertoli cells (SC) seen as lipid infiltration. The spermatogenic cells presented variable changes characterized by the disorientation of cells, maturation arrest, and premature sloughing. No immune complex deposits were seen. Of the 9 cases followed, pregnancy occurred in 2 following vas reanastomosis; a regenerative capacity of the ST was seen in 6 cases. The follow-up studies also show that the morphological alterations initially produced by the vasectomy have little affect on the appearance of spermatogenesis after vas reanastomosis. Thus, we see that the vasectomy exerts its major effect on the SC and that the damage to these cells alters the testicular environment. However, these changes are apparently reversible following vasovasostomy.     

Apoptosis and testicular alterations in albino rats treated with etoposide during the prepubertal phase

Etoposide is a podophyllotoxin semiderivative that is used in a variety of chemotherapy treatments, including therapy for children tumors. This drug promotes the formation of a ternary DNA-topoisomerase II-etoposide complex that triggers apoptosis. The purpose of this work was to analyze the occurrence of apoptosis in the seminiferous epithelium of prepubertal, pubertal, and adult rats treated with 10, 20, and 40 mg/Kg of etoposide during the prepubertal phase, as well as the role of apoptosis in etoposide-induced testicular damage. The rat testes were fixed in Bouin's liquid, and the apoptotic cells were quantified by means of the hematoxylin and eosin (H&E) technique (all groups) and the terminal dUTP nick end labeling (TUNEL) method (prepubertal groups only). The results obtained from both the H&E and TUNEL methods showed an increased frequency of apoptosis in the seminiferous epithelium of treated animals, except for the subgroup that received the 10-mg/Kg dose and was sacrificed 12 hr after the treatment and for the etoposide-treated pubertal group, that did not show cells suggesting apoptosis during H&E analysis. The labeled cells were mainly primary spermatocytes and differentiated spermatogonia. The prepubertal rats showed an etoposide-dose-dependent diminution of differentiated spermatogonia. Etoposide treatment during the prepubertal phase increases the frequency of apoptosis in the seminiferous epithelium, and causes serious harm to male fertility. 2004.     

输精管结扎术后附睾精子及其吻合术后再现精子超微结构的观察

目的观察输精管结扎术后附睾精子和吻合术后再现精子超微结构的演变情况及其相关关系。方法15例要求作输精管吻合术的对象,手术时取出附睾精子（A组）并追踪其吻合术后精液再现精子（B组）,同时应用光镜和电子显微镜分别观察A组、B组的精子形态和结构的改变,并进行统计学分析。结果光镜分析显示：B组与A组比较,精子密度和尾畸比例下降有显著性;而精子活动率和头畸比例升高有显著性;正常形态的比例有增高的趋势。电镜分析显示：B组与A组比较,正常形态精子显著升高,而尾部畸形精子明显降低,两者差异均有统计学的显著性;头畸形和颈畸形的比例在两组间差异没有显著性;而两组间尾部畸形比例呈正相关。透射电镜显示内部结构在头、颈、尾均有多细胞器、多种形式的改变。结论本文用电镜观察进一步证实了输精管结扎术后附睾精子形态向输精管吻合术后精液再现精子变化的规律。不论电镜还是光镜分析,精子尾部异常都是由高比例（A组）向低比例（B组）转化,正常形态的精子均由低比例（A组）向高比例（B组）转化;精子头部的异常光镜分析是一个由低比例（A组）向高比例（B组）转化的过程,而电镜分析没有改变。本文在A组和B组未有发现与输精管结扎术有关的精子形态学上特征性的改变。     

Distribution of Zonula Occludens‐1 and Occludin and alterations of testicular morphology after in utero radiation and postnatal hyperthermia in rats

In utero irradiation (IR) and postnatal hyperthermia (HT) exposure cause infertility by decreasing spermatogenic colony growth and the number of sperm in rats. Four groups were used: (i) Control group, (ii) HT group (rats exposed to hyperthermia on the 10th postnatal day), (iii) IR group (rats exposed to IR on the 17th gestational day) and (iv) IR + HT group. Three and six months after the procedures testes were examined by light and electron microscopy. Some degenerated tubules in the HT group, many vacuoles in spermatogenic cells and degenerated tight junctions in the IR group, atrophic tubules and severe degeneration of tight junctions in the IR + HT group were observed. ZO‐1 and occludin immunoreactivity were decreased and disorganized in the HT and IR groups and absent in the IR + HT group. The increase in the number of apoptotic cells was accompanied by a time‐dependent decrease in haploid, diploid and tetraploid cells in all groups. Degenerative findings were severe after 6 months in all groups. The double‐hit model may represent a Sertoli cell only model of infertility due to a decrease in spermatogenic cell and alterated blood‐testis barrier proteins in rat.     

The effect of immunomodulating drugs on adjuvant-induced arthritis in Lewis rats

The purpose of this study was to investigate the effects of cyclosporine A (CSA) and methotrexate (MTX) as potential immunomodulators in a nonestablished adjuvant arthritis (AA) model. Non-injected hind paw volumes were reduced when AA rats were treated for 18 dayswith CSA (100% at 10 mg/kg) or MTX (100% at 0.1 mg/kg). Body weights of drug treated AA rats were increased above untreated AA rats and were similar to non-arthritic controls. AA rats show elevated T helper (W3/25⁺)/T suppressor (OX 8⁺) cell ratios (2.0 vs. 3.1,p<0.01). The immunomodulators tested all returned these elevated ratios to control non-arthritic levels. Similarly, these drugs returned the reduced mitogen responses and elevated blood granulocyte numbers toward normal non-arthritic control values.     

The Ghana vasectomy initiative: facilitating client-provider communication on no-scalpel vasectomy

Objective

In 2003-2004 and 2007-2008, an initiative was implemented to improve client and provider knowledge and acceptance of no-scalpel vasectomy (NSV) in Ghana.

Methods

At eight facilities, physicians were trained in NSV and staff received training in the provision of “male-friendly” services. Health promotion activities provided NSV information to prospective clients. Client-provider communication was assessed via a mystery client study (n = 6). Knowledge and acceptance of NSV among potential clients were assessed with baseline and follow-up surveys (each n = 200) in 2003-2004 and three follow-up panel surveys in 2008 (each n = 240).

Results

Trained health staff exhibited improved attitudes and knowledge regarding NSV. Mystery clients reported receiving accurate, nonjudgmental NSV counseling. Awareness of NSV among panel respondents doubled from 31% to 59% in 2003-2004 and remained high (44%) in 2008. The proportion of men who would consider NSV increased from 10% to 19% in 2007-2008. NSV procedures increased three-fold from 2003 (n = 26) to 2004 (n = 83) and 2007 (n = 18) to 2008 (n = 53).

Conclusion

Provider training in client-centered services, coupled with targeted health promotion, improved client and provider knowledge and acceptance of NSV in an African context.

Practice implications

Complementary, sustained provider training and health promotion are needed to maintain NSV service quality and acceptance.     

Hepatocellular alterations after intraportal transplantation of ovarian tissue in ovariectomized rats

The mechanisms of hepatocarcinogenesis by certain synthetic estrogens seem to involve both nongenotoxic and indirect genotoxic effects. However, the natural estrogen estradiol did not exert any carcinogenic effects in established experimental protocols. To elucidate specific long-term effects of natural estrogens on hepatocytes, small pieces of ovarian tissue were transplanted via the portal vein into the livers of ovariectomized female rats. One week, 3 weeks, and 3 months after transplantation the transplants were found to proliferate and to secrete estradiol. Three weeks after transplantation the hepatocytes of the liver acini downstream of the stimulated transplants already showed a remarkable loss of glycogen, distinct cytoplasmic amphophilia, enlargement of their nuclei, a strong increase in the number and size of peroxisomes, an increase in proliferative activity and apoptotic elimination, and changes in the activity of certain key enzymes of energy metabolism. All hepatocellular alterations could be inhibited by the estrogen receptor antagonist toremifene and are, therefore, attributed to specific effects of estradiol produced by the transplants. The observed alterations resemble in some respects amphophilic preneoplastic liver foci, which particularly occur after long-term administration of nongenotoxic hepatocarcinogens, including the adrenal steroid hormone dehydroepiandrosterone. In a preliminary experiment three of six animals exhibited a hepatocellular carcinoma, and another animal developed a hepatocellular adenoma 18 months after intrahepatic ovarian tissue transplantation.     

Ultrastructural alterations in hexafluoroacetone-induced testicular atrophy in the rat

Testicular atrophy was induced in rats by dermal application of hexafluoroacetone (HFA) at 39 or 130 mg/kg/day for 14 days, but not at a dosage of 13 mg/kg/day. Affected germ cells were mostly spermatids and to a much lesser extent spermatocytes; spermatogonia were unaffected. Late spermatids were retained in Sertoli cells and showed degenerative changes. Sertoli cells exhibited cytoplasmic vacuolation, distended endoplasmic reticulum, and a marked increase in lipid droplets. Leydig cells exhibited a slight increase in lipid droplets, fewer mitochondria, and diminution and segregation of the agranular endoplasmic reticulum from mitochondria. A correlation between ultrastructural and biochemical changes in HFA-induced testicular atrophy is presented.     

The effect of unilateral vasectomy on testosterone and testicular parameters in the adult male African giant rat (Cricetomys gambianus)

Background

The effects of vasectomy on spermatogenesis and reproductive parameters are recognized to be specie-dependent with marked differences in levels of perturbations observed.

Objectives

To assess the impact of unilateral vasectomy on testosterone level and other testicular parameters in the male African giant rat (AGR) (Cricetomys gambianus).

Methods

Sixteen adult male AGRs weighing 500–1300 g were recruited for the experiment. Animals were randomly divided into three experimental groups (1–3) and one control (sham operated) group with four rats per group. Experimental vasectomy was done by carefully ligating the vas deferens of the right testis of all the experimental groups (1, 2, and 3) and animals were allowed either 8, 6 and 2 weeks respectively before sacrifice. Sham-operated animals served as the control. Blood samples were collected and assayed for testosterone while testicular tissue was further processed for seminal fluid and histo-pathological analyses.

Results

Spermatogenic parameters indicate a pattern of decline in sperm count and motility between the experimental groups and the control and azoospermia in the eight-week group. Histological alterations were marked by atrophy of seminiferous tubules which was proportional to the duration of vasectomy. Serum testosterone levels were significantly reduced at eight weeks. There was no statistically significant difference between sperm counts of right and left testes except for group 3. Results suggest that unilateral vasectomy of the AGR may have negative impact on the contralateral testis in the male African giant rat.

Conclusion

These preliminary results reveal that unilateral vasectomy in the AGR may result in perturbations of the histo-architecture of the testes with possible decline in function.