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1.

Background

During epididymal transit, functional and structural modifications leading to full maturation enable male gametes to reach, recognize and fertilize the oocytes. In dogs, little is known on the modifications of spermatozoa during the passage in the epididymis. The aim of this study was to describe the motility, morphology and acrosomal patterns of canine spermatozoa retrieved from the epididymis caput, corpus and cauda.

Results

After the dilution required for the collection of epididymal content, sperm motility was significantly higher (P <0.0001) in the cauda compared to corpus and caput.Proportions of spermatozoa with normal morphology were significantly higher in corpus (P =0.02) and cauda (P <0.0001) compared to caput. Overall morphological abnormalities of the head and neck/midpiece were similar in the three different epididymal regions. A significantly increased prevalence of tail defects, mainly represented by single bent tails, was observed in the corpus compared to caput (P <0.0001) and cauda (P =0.006).Numbers of immature sperm with cytoplasmic droplets decreased from the proximal to the distal region of the epididymis. Particularly, proximal cytoplasmic droplets were more frequently found in spermatozoa collected from the caput epididymis than in the corpus (P <0.0001) and in the cauda (P <0.0001), whereas the occurrence of distal cytoplasmic droplets was higher in the corpus than in the caput (P =0.0003) and in the cauda (P <0.05).Significantly higher proportions of spermatozoa with intact acrosomes were retrieved from the cauda epididymis than from the caput (P =0.03) and the corpus (P =0.008). This difference was mainly due to a lower proportion of spermatozoa with abnormal acrosomes (mainly swollen acrosomes) rather than with absent acrosomes.

Conclusions

Canine spermatozoa undergo several modifications in the epididymis. The acquisition of progressive motility, migration of the cytoplasmic droplet and acrosomal reshaping lead to mature spermatozoa which are then stored in the cauda epididymis. From this site, spermatozoa can be retrieved and used in assisted reproductive techniques as a valuable tool for propagating genetic traits of high value individuals that dies accidentally or undergoes orchiectomy for medical purposes. Further investigations should be also focused on the potential use of spermatozoa recovered from other epididymal regions.  相似文献   

2.
哺乳动物精子经过附睾成熟后才能获得运动及受精的能力,为解释水牛精子在附睾中的成熟过程,本研究选用性成熟期的沼泽型水牛附睾,利用乙烯吡咯烷酮包裹的硅胶微小颗粒(Percoll)梯度离心纯化分别提取附睾头、体和尾部精子,应用计算机辅助精子分析系统(CASA)检测精子活力,透射电镜观察附睾不同部位精子的超微结构,对精子进行荧光标记后,利用流式细胞仪和荧光显微镜观察检测不同部位精子质膜完整率、线粒体鞘膜电位和顶体差异。结果表明,Percoll分离得到附睾头、体和尾3部位精子的纯度达95%,不同部位精子活力分别为8.35%、20.21%和65.60%;附睾不同部位精子都存在着结构完整的精子以及相同的畸形类型,附睾尾部精子线粒体鞘高膜电位比率最高,精子质膜完整率从附睾头部到尾部逐渐升高,精子顶体完整率从附睾头部到尾部逐渐升高。本研究直观地展示了水牛附睾不同部位精子特征以及差异,为研究水牛精子成熟机理提供理论依据。  相似文献   

3.
Semen samples collected postmortem from 142 yearling beef bulls (11-13 months old) of three different breeds (Charolais, Hereford and Simmental) were examined to evaluate the proportion of bulls with mature spermiograms. Before slaughter, testes and epididymides were clinically examined and scrotal circumferences were measured. Aliquots of the cauda epididymal contents taken postmortem were used for sperm morphology examination. Sperm head morphology was studied in dry smears stained with carbol-fuchsine. For each preparation, 500 spermatozoa were counted in each smear under light microscope (x 1000). The presence of proximal cytoplasmic droplets, abnormal acrosomes, detached heads and abnormalities of the midpiece and tail were recorded in wet preparations of formol-saline-fixed spermatozoa. For each preparation, 200 spermatozoa were counted in each preparation under a phase-contrast microscope (x 1000). The abnormalities were classified according to a classification system developed by Bane (1961). Morphological abnormalities were recorded as a percentage of the total number of counted spermatozoa. Criteria for a spermiogram to be considered mature included <15% abnormal heads and <15% proximal droplets. According to this definition approximately 48% (68 of 142) of the examined bulls were considered mature. The bulls in this study represent approximately one-fifth of the total amount of performance-tested beef bulls in Sweden during 5 years. Our results indicate that only less than half of the Swedish yearling beef bulls at the testing station appear to have a mature spermiogram at the time they are offered for breeding purposes.  相似文献   

4.
The appearance and incidence of sperm abnormalities was studied in 115 ejaculates, collected periodically over 1 year covering all seasons from five mature, healthy swamp buffalo (Bubalus bubalis) bulls reared under tropical conditions and serving as the current source of semen for artificial insemination (AI) in Thailand. Light microscopy of stained smears was used to investigate sperm head shape morphology, while unstained wet smears were used to examine other sperm abnormalities. The most commonly found morphological aberrations were pear-shaped spermatozoa, knobbed acrosomes, proximal cytoplasmic droplets, simple bent tails and coiled tails under the head, whose ultrastructure (scanning electron microscopy) corresponded to what has been found in other species of bovidae, including varieties of buffalo. The mean prevalence (as least squares mean +/- SEM) of sperm abnormalities was low (below 15%), corresponding to healthy spermiograms. The younger bulls (<10 years old, n = 3) had less abnormalities than the older ones (10.1 +/- 0.6% versus 14.1 +/- 0.8%, P < 0.001, n = 2), including abnormalities of sperm head shape (1.1 +/- 0.3% versus 3.6 +/- 0.3, P < 0.001), acrosome defects with knobbed acrosomes (1.1 +/- 0.2% versus 1.2 +/- 0.3%, P < 0.001), spermatozoa with proximal cytoplasmic droplets (2.7 +/- 0.1% versus 1.4 +/- 0.2%, P < 0.001), defective mid-pieces (0.2 +/- 0.1% versus 0.3 +/- 0.1%) and abnormal sperm tails (3.1 +/- 0.3% versus 5.7 +/- 0.4%, P < 0.001). The within-bull effect of the year solely affected the incidence of pear-shaped spermatozoa while the incidences of abnormal contour, variable size of sperm head shapes, abnormal mid-piece and simple bent tail among bulls were affected by ejaculate (week of collection). Interaction between age and ejaculate affected only the prevalence of spermatozoa with proximal cytoplasmic droplets. In conclusion, the types of defects encountered were similar to those found in other bovidae, with a very low prevalence over the year the AI sires were followed through.  相似文献   

5.
Seventeen gonad pairs of boars and ten gonad pairs of bulls were examined to evaluate the migration of protoplasmic droplets and the phagocytosis of defective spermatozoa. The material for a microscopic investigation of secretions was collected from two sites in the testis and from seven sites in the epididymis. The greatest motion of protoplasmic droplets was recorded in the caput epididymidis, although the migration of droplets from the proximal section of the connective part of the flagellum towards the distal parts could also be observed as far as in the cauda epididymidis in both animals. A proximally located droplet still occurred in the cauda epididymidis in 4.5% of the spermatozoa of boars and in 1.9% of those of bulls. Absent mitochondrial spirals or swollen connective parts were observed in the imprints of testicular tissue in almost 50% of the spermatozoa whereas in the secretion of efferent ducts they were observed only in 0.3% of bull spermatozoa and about 3% of boar spermatozoa. No such defects were recorded in the epididymis head and tail in either of the two species. The marked reduction in the number of defective spermatozoa without mitochondrial spirals in the secretion of efferent ducts and after passage through the caput epididymidis testifies to the phagocytic ability of the epithelium of this part of efferent ducts.  相似文献   

6.
Epididymal spermatozoa from moose were studied in phase contrast, light interference and electron microscope. Some samples taken from cauda were diluted and frozen in liquid N2. The motility of the sperms after thawing was good.The concentration of spermatozoa in cauda was calculated to 10 × 106 cells per µl.Morphologically the spermatozoa of moose were found to be quite similar to those collected from bulls. The length of the sperm head was found to be approx. 8.8 µ and the average maximal width 5.2 µ. The average length of the tail was 54.7 µ and the entire length of the spermatozoon varied from 60 to 64 µ. Compared with sperm cells from bulls the moose spermatozoa appeared to have a somewhat shorter and broader head and a slightly shorter tail.The migration of the cytoplasmatic droplets, which was found to be completed in caput, seemed to follow the same pattern as in bulls and boars. As found in these species there was also in the moose a higher frequency of secondary abnormalities in the spermatozoa from cauda than in those from the other parts of epididymis.Studies of the fine structure of the moose spermatozoa seemed to indicate that these are of the same type as the spermatozoa of bulls, rams and boars. In sagittal sections the sperm head was thin, but in contrast to the sperm cells of the species mentioned above no typical waist-like narrowing in the equatorial region was found. The equatorial segment also seemed to be less arched than in the spermatozoa from bulls, rams and boars. Otherwise, no principal difference was found between ultrastructure of the moose spermatozoa and that of the spermatozoa collected from domestic species.  相似文献   

7.
To clarify the physiological changes of sperm morphology in active Thoroughbred stallions during the breeding season, we examined the dismount semen collected from the penile urethra immediately after service. The spermatozoa were analyzed for relationships between the morphology and the stallion’s age or the number of services. Seasonal variation was apparent in the rate of the sperm tail abnormalities, spermatozoa with cytoplasmic droplets, appearance of medusa cells, and sperm head length. Area and width of the sperm head correlated negatively with age (P<0.05). The rate of appearance of medusa cells and the length of the sperm head were positively related to the number of services (P<0.05), and the aspect ratio was negatively related (P<0.01).  相似文献   

8.
Three experiments evaluated the effects of dietary Se and vitamin E on the ultrastructure of spermatozoa, ATP concentration of spermatozoa, and the effects of adding sodium selenite to semen extenders on subsequent sperm motility. The experiment was a 2 x 2 arrangement of treatments in a randomized complete block design. A total of 10 mature boars were fed from weaning to 18 mo of age diets fortified with two levels of supplemental Se (0 or .5 ppm) or vitamin E (0 or 220 IU/kg diet). The nonfortified diets contained .06 ppm Se and 4.4 IU vitamin E/kg. In Exp. 1, the spermatozoa from all boars were examined by electron microscopy. Vitamin E had no effect on structural abnormalities in the spermatozoa. When the low-Se diet was fed the acrosome or nuclei of the spermatozoa was unaffected, but the mitochondria in the tail midpiece were more oval with wider gaps between organelles. The plasma membrane connection to the tail midpiece was not tightly bound as when boars were fed Se. Immature spermatozoa with cytoplasmic droplets were more numerous when boars were fed the low-Se diet, but the occurrence of midpiece abnormalities occurred in boars fed diets with or without Se or vitamin E. Our results suggest that Se may enhance spermatozoa maturation in the epididymis and may reduce the number of sperm with cytoplasmic droplets. In Exp. 2, the concentration of ATP in the spermatozoa was evaluated in the semen of all treatment boars. When the low-Se diet was fed, ATP concentration was lower (P < .01), whereas vitamin E had no effect on ATP concentration. Experiment 3 investigated the effect of diluting boar semen with a semen extender with sodium selenite added at 0, .3, .6, or .9 ppm Se. Three ejaculates from each boar were used to evaluate these effects on sperm motility to 48 h after dilution. Sperm motility declined (P < .01) when Se was added to the extender, and this decline was exacerbated as the concentration of added Se increased (P < .01). The added Se was demonstrated to be tightly adhered to the spermatozoa. Overall, these results suggest that low Se-diets fed to boars resulted in abnormal spermatozoal mitochondria, a lower ATP concentration in the spermatozoa, and a loose apposition of the plasma membrane to the helical coil of the tail midpiece, but no effect from inadequate vitamin E was demonstrated. Adding sodium selenite to the semen extender reduced sperm cell motility.  相似文献   

9.
On assessment for use in an AI stud, a 12‐month‐old bull was found to produce low volume ejaculates with 41% of the sperm having morphological abnormalities. No left epididymal tail was palpable and the head of the epididymis on the left was twice the size compared with the right. Ultrasound examination showed the left testis to contain a large central area of decreased echogenicity, which could be followed proximally to a 15‐mm echolucent lesion at the site of the epididymal head. Postmortem examination revealed a 15‐mm diameter cyst in the region of the left epididymal head, and absence of the body and tail of the epididymis. The mediastinum testis of the left testis was dilated, corresponding to the area of decreased echogenicity observed on ultrasonography. No left seminal vesicle was present and the ampulla was significantly smaller than the same structure on the right. Histological examination revealed incomplete or absent spermatogenesis involving the majority of seminiferous tubules in the left testis, and a small proportion of those of the right testis. The cystic structure at the site of the left epididymal head was lined by irregular, sometimes attenuated, epithelium and contained sparse spermatozoa. This case demonstrates the adverse impact, which segmental aplasia of the mesonephric duct had on the testicular and epididymal function of a bull, and highlights the importance of careful clinical assessment in its diagnosis.  相似文献   

10.
The effectiveness of the water test, short hypoosmotic swelling test with ultrapure water was examined in canine epididymal spermatozoa to evaluate tail membrane integrity. Spermatozoa during epididymal transit were also characterized. Sperm suspension obtained from cauda epididymis was diluted 1:4 with ultrapure water, and incubated for 5 min. The percentage of swollen spermatozoa in the water test was significantly correlated with both the sperm motility and the swelling value obtained by the conventional hypoosmotic swelling test. Canine spermatozoa collected from the caput epididymis were not motile, but revealed membrane integrity in a water test. The water test can be used as a simple and short hypoosmotic swelling test to evaluate the tail membrane integrity of canine epididymal spermatozoa.  相似文献   

11.
The morphology of spermatozoa of modern Thoroughbred stallions in Japan was investigated during the breeding season. A total of 299 semen samples were collected from the penises of 16 stallions immediately after service. The rate of abnormalities in sperm heads and tails, spermatozoa with cytoplasmic droplets and slides with medusa cells to total observed slides in each stallion were 3.9 +/- 2.1%, 11.5 +/- 5.9%, 2.4 +/- 2.6% and 20.1%, respectively. The values for the area, length, width and aspect ratio of the stallion sperm head were 12.54 +/- 1.34 microm(2), 5.93 +/- 0.40 microm, 2.69 +/- 0.21 microm and 0.46 +/- 0.05, respectively. With the exception of medusa cells, the features were significantly different among the stallions (P<0.05).  相似文献   

12.
Tail and mid-piece morphology of ram spermatozoa were compared using wet preparations of semen diluted in buffered formal saline at temperatures of 10 degrees C and 65 degrees C. The temperature of the diluent did not affect the occurrence of abnormalities. Tail and mid-piece morphology were also examined in semen smears stained by a modified Williams' stain using glass slides at temperatures of 10 degrees C, 38 degrees C and 65 degrees C. The occurrence of the abnormalities was not affected by the slide temperature. The occurrence of tail and mid-piece abnormalities was compared using the 2 methods of preparation. Only the occurrence of distal cytoplasmic droplets was affected by the method of preparation. In the stained smear preparations, most of the distal cytoplasmic droplets were lost. However, only a few of the proximal droplets were lost when this method was used.  相似文献   

13.
The aim of this study was to investigate the effect of season, temperature, humidity, age of the boar, and semen collection interval on sperm morphology in Duroc boars in Thailand, kept either in a conventional open air system (CONV) or in an evaporative cooling system (EVAP). In total, 1176 ejaculates from 110 sexually mature boars in six CONV herds and five EVAP herds were morphologically examined during a one-year period. Analysis of variance was applied to the data. Minor differences in the sperm morphology traits analyzed were found between the housing systems. There was a significant seasonal effect (two-month periods) on the percentage of morphologically normal spermatozoa (normal1), morphologically normal spermatozoa including spermatozoa with distal cytoplasmic droplets (normal2), proximal cytoplasmic droplets (prox), and sperm head abnormalities (P相似文献   

14.
Sperm morphology was studied in 302 extensively managed Zebu bulls (aged 1.5–9 years), classified as sound (n=166) or unsound (n=136) for breeding, under field conditions in the dry tropics of Costa Rica. Single semen samples were collected by electro‐ejaculation and fixed in formol‐saline solution immediately after collection. Sperm morphology was determined in the field on wet smears using a microscope equipped with phase‐contrast optics, and further determined in the laboratory on air‐dried smears stained with carbol‐fuchsin. The frequencies of sperm abnormalities (such as abnormal acrosome, head, neck, mid‐piece, tail, and presence of cytoplasmic droplets) were recorded as a percentage of the total number of counted spermatozoa (400 cells). Zebu bulls considered unsound for breeding showed a higher mean prevalence (p < 0.05) of knobbed acrosomes (4.0 versus 0.9%), head defects [specifically, nuclear invaginations and heads with abnormal shapes and sizes (27.6 versus 4.0%)], abnormal tails (11.2 versus 4.7%), and proximal droplets (8.4 versus 1.6%), compared with bulls considered sound for breeding. In these latter bulls, the abnormality most commonly seen was the presence of single bent tails with an entrapped cytoplasmic droplet (3.0 ± 3.7%). Young Zebu bulls (i.e. bulls under 2 years of age) showed a higher percentage of missing acrosomes, and proximal cytoplasmic droplets, than older sires (12.1 versus 2.4%, and 23.9 versus 3.6%, respectively; p < 0.05), interpreted as an indication of low ejaculation frequency and sexual immaturity, respectively. Bulls with a long scrotum and soft testicular consistency (TC) at palpation showed higher percentages of abnormal sperm heads in the ejaculate than bulls with a normal scrotal length (SL) and a normal TC (32.7 versus 12.8% and 30.7 versus 10.3%, respectively; p < 0.05). In addition, Zebu bulls with a scrotal circumference (SC) ≤ 30 cm showed a higher prevalence of proximal cytoplasmic droplets than bulls whose SC was > 30 cm (9.8 versus 2.6%, p < 0.05). A higher mean percentage of abnormally sized and shaped heads, especially undeveloped and narrow at the base, was more frequently found in stained smears than in unstained samples (26.0 versus 9.9%, p < 0.05), which clearly underlines the importance of using both stained and wet smears when assessing sperm head morphology. However, for a quick assessment of sperm morphology under field, tropical conditions, phase‐contrast microscopy provides useful information for the spermiogramme evaluation.  相似文献   

15.
Spermatozoa and developing spermatids showing neck region abnormalities have been studied in material from 2 genetically related boars. In both boars the defects were abaxial implantation of the tails and lack of substance in the neck region. In many spermatozoa, a too wide space between the capitulum and the basal plate was more pronounced in epididymal spermatozoa compaired to testicular material. This implies that the defect aggravated, and might be connected with the migration of the cytoplasmic droplet in the epididymis. Since the defects were observed in spermatids, it its concluded that the defects were heriditary.This conclusion was further supported by the observation of similar defects in 6 other related boars, examined by light microscopy only.  相似文献   

16.
The influence on sperm morphology of different methods for preparation of semen and of storage in a fixative solution was examined in 27 beef bulls subjected to a regular breeding health examination. Sperm head morphology under light microscopy did not differ between smears of fresh semen stained with carbol-fuchsin-eosin (Williams staining) or Nigrosin-Eosin. Nor was there any difference between samples stained immediately after collection and those stained after 1 month of storage at + 4 degrees C in buffered formal-saline solution. Formol-saline fixed spermatozoa examined in wet preparations under phase contrast microscopy had a higher prevalence of acrosome defects and cytoplasmic droplets than stained smears of fresh semen under light microscopy. One month of storage in formol-saline did not affect the prevalence of acrosome defects or cytoplasmic droplets. There was no influence of fixation method (wet or dry), staining, examination technique, or storage time on midpiece or sperm tail morphology. The affinity of spermatozoa to eosin at staining with Nigrosin-Eosin ("live and dead count") did not differ between fresh semen and spermatozoa that had been stored in formol-saline for 1 month. It is concluded that bull semen can be stored for at least 1 month at + 4 degrees C in buffered formal-saline without major changes in sperm morphology. Furthermore, examination of wet preparations of fixed spermatozoa under phase contrast microscope is likely to yield the most accurate results for morphological characteristics like acrosome morphology and cytoplasmic droplets.  相似文献   

17.
Feral raccoons (Procyon lotor) have been growing in number in Japan, and they are becoming a problematic invasive species. Consequently, they are commonly captured and killed in pest control programs. For effective population control of feral raccoons, it is necessary to understand their reproductive physiology and ecology. Although the reproductive traits of female raccoons are well known, those of the males are not well understood because specialized knowledge and facilities are required to study them. In this study, we first used a simple evaluation method to assess spermatogenesis and presence of spermatozoa in the tail of the epididymis of feral male raccoons by histologically examining the testis and epididymis. We then evaluated the possibility of using 7 variables—body weight, body length, body mass index, testicular weight, epididymal weight, testicular size and gonadosomatic index (GSI)—to estimate spermatogenesis and presence of spermatozoa in the tail of the epididymis. GSI and body weight were chosen as criteria for spermatogenesis, and GSI was chosen as the criterion for presence of spermatozoa in the tail of the epididymis. Because GSI is calculated from body weight and testicular weight, this model should be able to be used to estimate the reproductive state of male raccoons regardless of season and age when just these two parameters are known. In this study, GSI was demonstrated to be an index of reproductive state in male raccoons. To our knowledge, this is the first report of such a use for GSI in a member of the Carnivora.  相似文献   

18.
Sperm cells isolated from the tail of the epididymis and from the semen of the same individuals were analysed. The use of silver nitrate to stain sperm cells isolated from the tail of the epididymis made it possible to identify structures that were not visible in the sperm from semen. Silver nitrate very clearly distinguished the acrosomal and distal parts of the sperm head. Following silver nitrate staining, the sperm isolated from the tail of the epididymis were characterized by dark ‘collars’ in the distal part of the head. These ‘collars’ are not visible in the sperm cells isolated from semen. The results of the study indicate differences in the dimensions of sperm isolated from the tail of the epididymis and sperm in semen. Sperm isolated from the tail of the epididymis had smaller heads, despite their longer length, and had longer midpieces and tails than ejaculate sperm. Silver nitrate staining is a simple and fast technique. Silver nitrate makes it possible to identify the acrosome and post-acrosomal region of the sperm head and to clearly identify the midpiece. Therefore, it can be successfully used to supplement routine techniques for evaluating sperm morphology or as an independent technique.  相似文献   

19.
A complete diallel cross involving two rabbit sire lines (C and R) was carried out to estimate the crossbreeding genetic parameters of seminal traits. 2140 ejaculates from 153 males were analyzed. The traits studied were: presence of gel plugs (G), urine (U), and calcium carbonate deposits (CC), number of useful ejaculates (UE), pH, volume (V), mass and individual motility (Mm, Mi), useful Mi (UM), concentration (Cn), number of spermatozoa per ejaculate (TSE), percentage of viable spermatozoa (Vi), spermatozoa with normal apical ridge (NAR), normal spermatozoa (Nr), spermatozoa with morphological abnormalities of head (H), neck-midpiece (Nm), and tail (T), presence of proximal and distal cytoplasmic droplet (Dp, Dd).

Estimates of heterosis, direct and maternal genetic effects were obtained from the solutions of the mixed model. There were major differences in direct genetic effects between lines, which were favourable to line C for Cn and TSE, and unfavourable for CC, Nm and Dp. Smaller differences were also observed in Vi and NAR favourable to line R. Differences between lines with respect to the maternal genetic effects were relevant and favourable to line C for V and to line R for U, UM, Cn, TSE, Nm, Mi, and Mm. Individual heterosis was high for Dp and Dd.  相似文献   


20.
Microspectrophotometric, microinterferometric and microplanimetric techniques were used to investigate whether the maturation of bull spermatozoa is associated with any detectable changes in the major chemical components of the spermatozoal head. Round spermatids, caput, corpus, cauda epididymidal and ejaculated spermatozoa were obtained from six bulls with lowered fertility and three bulls with normal fertility. There was no quantitative change in DNA, in total dry mass or in optical area associated with the passage of spermatozoa between these areas of the epididymis. A marked reduction in relative Feulgen reactivity during spermiogenesis was found and was significantly more pronounced in spermatozoa from bulls with normal fertility than in spermatozoa from bulls of lowered fertility. It was suggested that this reduction signifies qualitative alterations of the spermatozoal deoxyribonucleoprotein complex and may be related to the fertility status of the bull.  相似文献   

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