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Syrian hamsters are photoperiodic rodents in which reproduction, including testicular function, is stimulated by long photoperiod exposure and curtailed by exposure to a short photoperiod. The objectives of this study were to characterize the testis histomorphometrically and to determine the role of the proliferation and apoptosis phenomena in the recovery of the seminiferous epithelium during spontaneous recrudescence after exposure to short photoperiod. The study was performed using conventional light microscopy, proliferating cell nuclear antigen and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP in situ nick end labelling staining, image analysis software, and transmission electron microscopy in three recrudescence groups: initial recrudescence (IR), advanced recrudescence (AR) and total recrudescence (TR). The results morphometrically pointed to the gradual recovery of the testicular and tubular volumes, as well as of the seminiferous epithelium. Among the IR and AR groups, the increase in testicular and tubular volumes was accompanied by an increase in tubular diameter and length, with an increase in interstitial volume. From AR to TR, there was an increase in the tubular and total volumes, but, in this case, with a gradual increase in tubular diameter. Recovery of the seminiferous epithelium was accompanied by changes in apoptosis and proliferation activities. The first decreased halfway through the process, and the second remained higher than the control levels throughout the recrudescence stage. Alterations in the spermatozoa were ultrastructurally observed, which indicated that spermiogenesis was not yet completely normal. In conclusion, spontaneous testicular recrudescence in Syrian hamster comprises two histomorphometrical phases: the first related to an increase in tubular length and diameter and interstitial volume and the second depending principally on the gradual increase in tubular diameter. The restoration of the seminiferous epithelium is due to apoptosis reaching normal values in the AR group accompanied by higher proliferative activity than that observed in the Control group., (© 2018 Blackwell Verlag GmbH.)

Yellowish myotis, Myotis levis, is a seasonal, epididymal sperm-storing Neotropical vespertilionid. In the dry season, males show simultaneous testis regression and sperm storage in cauda epididymis, enabling them to mate during this season. In this study, we investigated seasonal variations in body mass, diameter and height of seminiferous tubules and nuclei of Leydig cells in a population of southeastern Brazil. We also determined the frequencies of the stages of the seminiferous epithelium cycle (SEC) of mature individuals of this population. Body mass and diameter of Leydig cell nuclei showed no significant differences between dry and rainy seasons and stages of annual reproductive cycle; however, all other morphometric parameters varied significantly. The relative cumulative frequency of pre-meiotic stages of the SEC (1-3) was 51%, of meiotic stage (4) was 2% and of post-meiotic stages (5-8) was 47%. We confirmed that the yellowish myotis presents seasonal sperm production as revealed by testis regression and epididymal sperm storage during the dry season., (© 2014 Blackwell Verlag GmbH.)

The features of paca epididymis, based on its appearance in light microscope, is described in this paper. The cellular population of the epithelial lining comprises principal cells, basal cells, apical cells, narrows cells, and hallo cells. The epididymis is divided in five distinct and continuous regions, Zone I, or initial segment, and zone II, are both localized into the head. Zone III comprises the distal head and all the body. Zones IV and V are restricted to the tail, in the proximal and distal cauda epididymis respectively. Each zone can be readily distinguished on the basis of morphological characteristics. The height of epididymal epithelium is greater in zone I. There is a progressive increase in the diameter of the tubular lumen through the different areas, with the maximum in the zone V. The presence of a high epithelium, and the virtual absence of sperm in zone I suggest fast transit of spermatozoa in this region. Zone V comprises the distal tail, has smaller epithelial lining, greater luminal diameter, shorter stereocilia than the other zones, and contains spermatozoa packed inside the lumen, that characterizes this zone as a place of sperm storage. The findings are compared with other reports in rodents and other domestic animals, to contribute to the understanding of epididymal morphophysiology.

The seminiferous cycle of the rhinoceros was investigated for the first time using testicular tissue from captive white (Ceratotherium simum, n=2) and black (Diceros bicornis, n=1) rhinoceroses. Stages of the seminiferous epithelial cycle were characterised using the tubular morphology method and relative frequencies of each stage determined. This method allowed for the identification of eight stages of cellular associations characteristic of the seminiferous cycle in white and black rhinoceroses. The eight stages of the seminiferous cycle observed closely approximated the stages previously described for the domestic horse (Equus caballus). Premeiotic (stages I-III), meiotic (stage IV) and postmeiotic (stages V-VIII) represented 44.8%, 5.3% and 49.9% of the seminiferous cycle respectively., (Copyright © 2012 Elsevier B.V. All rights reserved.)

We describe seasonal variations of the histology of the seminiferous tubules and efferent ducts of the tropical, viviparous skink, Mabuya brachypoda, throughout the year. The specimens were collected monthly, in Nacajuca, Tabasco state, Mexico. The results revealed strong annual variations in testicular volume, stages of the germ cells, and diameter and height of the epithelia of seminiferous tubules and efferent ducts. Recrudescence was detected from November to December, when initial mitotic activity of spermatogonia in the seminiferous tubules were observed, coinciding with the decrease of temperature, photoperiod and rainy season. From January to February, early spermatogenesis continued and early primary and secondary spermatocytes were developing within the seminiferous epithelium. From March through April, numerous spermatids in metamorphosis were observed. Spermiogenesis was completed from May through July, which coincided with an increase in temperature, photoperiod, and rainfall. Regression occurred from August through September when testicular volume and spermatogenic activity decreased. During this time, the seminiferous epithelium decreased in thickness, and germ cell recruitment ceased, only Sertoli cells and spermatogonia were present in the epithelium. Throughout testicular regression spermatocytes and spermatids disappeared and the presence of cellular debris, and scattered spermatozoa were observed in the lumen. The regressed testes presented the total suspension of spermatogenesis. During October, the seminiferous tubules contained only spermatogonia and Sertoli cells, and the size of the lumen was reduced, giving the appearance that it was occluded. In concert with testis development, the efferent ducts were packed with spermatozoa from May through August. The epididymis was devoid of spermatozoa by September. M. brachypoda exhibited a prenuptial pattern, in which spermatogenesis preceded the mating season. The seasonal cycle variations of spermatogenesis in M. brachypoda are the result of a single extended spermiation event, which is characteristic of reptilian species., (Copyright © 2012 Wiley Periodicals, Inc.)

Unlabelled: The testis is a radiosensitive tissue. It contains a large number of lobules, which in turn are composed of convoluted seminiferous tubules. The epithelium inside each tubule consists of a complex mosaic of supporting cells and germ cells of different sizes and degrees of maturation. These cells are known to have diverse sensitivity to radiation, those with the highest sensitivity being the spermatogonia, which form part of the basal cell layer, and those with the lowest sensitivity being the mature sperm cells closest to the lumen of the tubule. For many years, the internal dosimetry community has discussed the need for improvements to bring about more detailed, cell-level testicular dosimetry. This paper presents a small-scale dosimetry model for calculation of S factors for several different source-target configurations within the testicular tissue., Methods: A model of the testis was designed in which the lobules were approximated by a cross-section of seminiferous tubules arranged in a hexagonal pattern, with interstitial tissue between them. The seminiferous tubules were divided into concentric layers representing spermatogenic development in the seminiferous epithelium. S factors were calculated for electrons, photons, α-particles, and for (18)F, (90)Y, (99m)Tc, (111)In, (125)I, (131)I, (177)Lu, and (211)At using Monte Carlo simulations., Results: For electrons with low energies the range was small, compared with the diameter of the seminiferous tubules, resulting in high energy deposition close to the source, whereas for higher electron energies more uniform energy deposition was seen, as expected. The same trend was seen for low-energy photons, whose mean free paths are small, compared with the diameter of the seminiferous tubules, resulting in high energy deposition close to the source, whereas for higher photon energies the location of the activity in the testis is less important., Conclusion: The model presented in this paper is a simplification of the organized chaos that constitutes the structure of the actual testis. However, it provides a relevant, small-scale anatomic model to help us understand the significance of the heterogeneity of radioactivity in this important radiosensitive tissue.

We performed histological analyses for comparing testicular microstructure between the gorilla, chimpanzee, and orangutan. Testicular samples were obtained by autopsy or biopsy from 10 gorillas, 11 chimpanzees, and 7 orangutans from several zoos and institutes. The seminiferous epithelia were thick in the chimpanzee and orangutan but thin in the gorilla. Leydig cells in the interstitial tissue were abundant in the gorilla. The acrosomic system was extremely well developed in the orangutans. Our study reveals that the cycle of seminiferous epithelium in orangutan testis can be divided into ten stages, whereas that in human, chimpanzee, and gorilla testes can be divided into only six stages. Phylogenetic analyses of the number of divisions may indicate that the seminiferous epithelium of our common ancestor has changed since the orangutan diverged from it. Furthermore, we performed comparative analyses of testicular microstructure to estimate relative sperm production among these three animals, and proposed a new indicator (namely the spermatogenic index, SI) closely related to sperm production. The SI indicated that a chimpanzee usually produces about 223 times more sperm than a gorilla and about 14 times more than an orangutan. Our data demonstrate the significance of the SI for estimating sperm production, thus aiding our understanding of the reproductive strategy as well as testis weight and relative testis size in investigated primates., (© 2011 Wiley-Liss, Inc.)

The radioprotective effects of granulocyte colony-stimulating factor (GCSF) were further investigated with respect to the testicular system. Recombinant human GCSF (100 μg kg(-1) body weight/day) was administrated to male C3H/HeN mice by subcutaneous injection for three consecutive days before pelvic irradiation (5 Gy) and histopathological parameters were assessed at 12 h and 21 days post-irradiation (pi). The GCSF protected the germ cells from radiation induced- apoptosis (P < 0.01 vs. irradiated group at 12 h pi). GCSF remarkably attenuated radiation-induced reduction in testis weight, seminiferous tubular diameter, seminiferous epithelial depth and sperm head count in the testes (P < 0.05 versus irradiated group at 21 days pi). Repopulation index and stem cell survival index of the seminiferous tubules were increased in the GCSF-treated group when compared with the radiation group (P < 0.01). The frequency of abnormal sperm in the GCSF group was lower than that in the irradiated group at 21 days pi (P < 0.01). The decrease in the sperm count and in sperm liability in the epididymis caused by irradiation was counteracted by GCSF. The present study suggests that GCSF protects from radiation-induced testicular dysfunction via an anti-apoptotic effect and recovery of spermatogenesis., (© 2010 Blackwell Verlag GmbH.)

Objective: This study aimed to determine the influence of high-dose soy isoflavones (daidzein and genistein) administered from prenatal life to sexual maturity on testosterone and estradiol levels, testicular and epididymal morphology, the number of epididymal spermatozoa, and mineral metabolism in rats., Methods: Pregnant Wistar rats received orally soy isoflavones, daidzein, and genistein at a dose of 200 mg/kg of body weight per day. After separating sucklings from their mothers, male rats received the same dose of isoflavones until reaching the age of sexual maturity, i.e., for 3 mo., Results: In the isoflavone-treated group, statistically significant decreased concentrations of zinc (determined using atomic absorption spectrophotometry) in blood serum and increased concentrations in bone were observed. The isoflavones induced changes in the morphology of the seminiferous epithelium of rat testes. However, there were no significant changes in the number of spermatozoa in the epididymis. The levels of estradiol in serum and cauda epididymis homogenates of rats receiving phytoestrogens were significantly higher than in the control group. No differences were observed in testosterone concentrations in the serum of treated and control rats. The testosterone levels in the homogenates of the treated rat testes were significantly lower than in the control group., Conclusion: The relatively mild effects of phytoestrogen administration on the morphology of testes and epididymides and the number of epididymal spermatozoa were observed despite the high dose used. The exposure of rats to genistein and daidzein during intrauterine life until sexual maturity influenced the mineral metabolism of the organism by significant decreases of Zn concentration in serum and increased Zn concentration in bones., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Similar to most wild felids, the ocelot (Leopardus pardalis) is an endangered species. However, knowledge regarding reproductive biology of the ocelot is very limited. Germ cell transplantation is an effective technique for investigating spermatogenesis and stem cell biology in mammals, and the morphologic characterization of germ cells and knowledge of cycle length are potential tools for tracking the development of transplanted germ cells. Our goal was to investigate basic aspects related to testis structure, particularly spermatogenesis, in the ocelot. Four adult males were used. After unilateral orchiectomy, testis samples were routinely prepared for histologic, stereologic, and autoradiographic analyses. Testis weight and the gonadosomatic index were 11+/-0.6g and 0.16+/-0.01%, respectively, whereas the volume density of seminiferous tubules and Leydig cells was 83.2+/-1.6% and 9.8+/-1.5%. Based on the acrosomic system, eight stages of spermatogenesis were characterized, and germ cell morphology was very similar to that of domestic cats. Each spermatogenic cycle lasted 12.5+/-0.4 d, and the entire spermatogenic process lasted 56.3+/-1.9 d. Individual Leydig cell volume was 2522mum(3), whereas the number of Leydig and Sertoli cells per gram of testis was 38+/-5x10(6) and 46+/-3x10(6). Approximately 4.5 spermatids were found per Sertoli cell, whereas daily sperm production per gram of testis was 18.3+/-1x10(6), slightly higher than values reported for other felids. The knowledge obtained in this study could be very useful to the preservation of the ocelot using domestic cat testes to generate and propagate the ocelot genome.

The ductus epididymis has roles in the maturation and storage of spermatozoa. The main function of the cauda epididymis is the storage of spermatozoa; however, this region exerts other morphophysiological roles. So, this study was aimed at investigating structural features of the cauda epididymis epithelium, which could indicate roles other than the storage. The relative percentages of the cell types in the epithelium were 74.9, 6.9, 12.5 and 5.6% of principal, clear, basal and halo cells respectively. Large intercellular spaces were seen among the lateral plasmatic membranes of adjacent principal cells or among these cells and others cell types. These spaces were found to be filled with multivesicular bodies, myelin figures, scrolls and debris of membranes or flocculent dense material. Clear cells had the cytoplasms filled with lysosomes ((3/4) of basal cytoplasm), and vacuoles and vesicles ((1/4) of apical cytoplasm). The observations allowed us to infer that clear cells could act in the process of endocytosis and also in water transfer from the lumen to the interstitium through the epithelium compartment. Moreover, transcytosis may occur at the cauda epididymis of Golden hamster.

The research aimed to study the morphologic variation of the testis, seeking to promote the selection and genetic control of those that present appreciable spermatic production throughout the year. Testis morphology of the Rynchotus rufescens partridge was investigated, analyzing the testis weight, the seminiferous tubules diameter, the thickness of the seminiferous epithelium, the amount of meiotic figures and the thickness of the tunica albuginea. Sixty male partridges were used, divided in 12 groups, and one group per month had the testis collected for the histological routine and the sections were stained using the Hematoxilin-Eosin technique. For the histological sections analysis, morphometric measures were taken, with the aid of an Image Analyzer and the resulting data were submitted to analysis of variance and to Tukey's test. Based on the histological modifications of the seminiferous epithelium and the morphometric analysis, the partridge testis morphology could be divided in four successive phases throughout the year. The reproductive phase occurred in the spring, characterized by the complete spermatogenesis process. The regression phase occurred in the summer, with the involution of the seminiferous epithelium. The rest phase took place in the fall, with spermatogonias presence and some spermatocytes beginning the meiosis. The phase of recrudescence occurred in the winter, with the recovery of the seminiferous epithelium and absence of spermatozoa. In conclusion, the characteristics analyzed revealed a variation over the year, with greater production of spermatozoa in the spring and less in the winter.

The objectives of this study were to identify and quantitate the germ cell populations of the testes in sexually mature male turkeys (Trial 1), determine the duration of meiosis based on BrdU labeling and stereological analyses (Trial 2), and examine the impact of various photoperiods on germinal and somatic cell populations in immature and adult males (Trial 3). In Trial 1, both testes within a male had similar stereological components (P>0.05) for all parameters analyzed. In Trial 2, the duration of Type-1 spermatocytes and round spermatids in turkeys lasted 4.5+/-0.5 and 2.0+/-0.5 days, respectively. In Trial 3, the short photoperiod (7L:17D) delayed testicular growth (in the stereological parameters analyzed). In contrast, the effect of a moderately short photoperiod (10.5L:13.5D) was comparable to the effect of a long (14L:10D) or increasing photoperiod (7L:17D to 14L:10D) on the stereological parameters examined. With the exception of the short photoperiod, all other photoperiods used in this study induced comparable early testicular maturation, with maximum testis weight at 29-35 weeks of age. As the males got older, there was a progressive, linear decline in testis weight through 60 weeks, at which time there were no significant differences among photoperiods. In conclusion, the duration of meiosis in the turkey was similar to that observed in the fowl and guinea-fowl. The existence of a threshold of photosensitivity to gonad stimulation in male turkeys is suggested to be between 7.0 and 10.5 h of light.

To clarify the reproductive state of male gorillas, we performed histological examinations on the testicles of 10 male gorillas (Gorilla gorilla). The testicular samples were obtained by autopsy, and ordinal histological preparations were made for light microscopy. The poor spermatogenesis of this species was characterized by the following findings: First, spermatogenesis was evident in only four samples. Meiosis progressed in two samples, but they lacked spermatogenesis. In the remaining four specimens, seminiferous tubules hyalinized without any sign of spermatogenesis. Second, seminiferous epithelia were thin even in the males in which spermatogenesis was observed. Third, degenerated seminiferous tubules were found in all specimens. Fourth, abnormally large syncytial cells were found in the tubules. Six stages in the epithelial cycle of the seminiferous tubules were identified. Testosterone staining made it clear that there were many Leydig cells with spherical or fusiform nuclei in an abundance of interstitial tissue. The relevance of the testicular architecture of gorillas to the mating system is discussed., (Copyright 2004 Wiley-Liss, Inc.)

Aim: To evaluate the effect of the alcoholic extract of Lepidium meyenii (Maca) on the spermatogenesis in male rats., Methods: In Holtzman rats, Maca alcoholic extract (5 %) was given by oral route at doses of 48 mg/day or 96 mg/day for 7 days, 14 days and 21 days. Testicular function was assessed by measurements of lengths of different stages of seminiferous epithelia and by epididymal sperm count., Results: Ethanolic extract of Maca increased the length of stages IX-XI of seminiferous epithelium at treatment day 7, day 14 and day 21. Progression of spermatogenesis was evident only after day 21 when lengths of stages XII-XIV of seminiferous epithelium were increased; at day 7 and day 14, no important change in spermatogenesis was observed. Epididymal sperm count was increased with 48 mg/day at all times. With 96 mg/day an increase in sperm count was observed at day 7, but it was reduced at day 14 and day 21 of treatment. Serum testosterone levels were not affected., Conclusion: The alcoholic extract of Maca activates onset ant progression of spermatogenesis at 48 mg/day or 96 mg/day in rats.

There is very little information regarding the testis structure and function in domestic cats, mainly data related to the cycle of seminiferous epithelium and sperm production. The testis weight in cats investigated in the present study was 1.2 g. Compared with most mammalian species investigated, the value of 0.08% found for testes mass related to the body mass (gonadosomatic index) in cats is very low. The tunica albuginea volume density (%) in these animals was relatively high and comprised about 19% of the testis. Seminiferous tubule and Leydig cell volume density (%) in cats were approximately 90% and 6%, respectively. The mean tubular diameter was 220 microm, and 23 m of seminiferous tubule were found per testis and per gram of testis. The frequencies of the eight stages of the cycle, characterized according to the tubular morphology system, were as follows: stage 1, 24.9%; stage 2, 12.9%; stage 3, 7.7%; stage 4, 17.6%; stage 5, 7.2%; stage 6, 11.9%; stage 7, 6.8%; and stage 8, 11 %. The premeiotic and postmeiotic stage frequency was 46% and 37%, respectively. The duration of each cycle of seminiferous epithelium was 10.4 days and the total duration of spermatogenesis based on 4.5 cycles was 46.8 days. The number of round spermatids for each pachytene primary spermatocytes (meiotic index) was 2.8, meaning that significant cell loss (30%) occurred during the two meiotic divisions. The total number of germ cells and the number of round spermatids per each Sertoli cell nucleolus at stage 1 of the cycle were 9.8 and 5.1, respectively. The Leydig cell volume was approximately 2000 microm3 and the nucleus volume 260 microm3. Both Leydig and Sertoli cell numbers per gram of testis in cats were approximately 30 million. The daily sperm production per gram of testis in cats (efficiency of spermatogenesis) was approximately 16 million. To our knowledge, this is the first investigation to perform a more detailed and comprehensive study of the testis structure and function in domestic cats. Also, this is the first report in the literature showing Sertoli and Leydig cell number per gram of testis and the daily sperm production in any kind of feline species. In this regard, besides providing a background for comparative studies with other fields, the data obtained in the present work might be useful in future studies in which the domestic cat could be utilized as an appropriate receptor model for preservation of genetic stock from rare or endangered wild felines using the germ cell transplantation technique.

The FHIT gene, located at human chromosome 3p14.2, is frequently deleted in a number of human cancers, and interstitial deletions at this site were recently described in a significant proportion (41%) of testicular germ cell tumors. We studied the expression of Fhit protein in the progression and differentiation of testicular germ cell tumors to further elucidate its role in this type of malignancy. Forty-five patients with testicular germ cell tumors and intratubular germ cell neoplasia (identified in 42/45 cases) were included in the study. Immunohistochemical staining with polyclonal rabbit IgG antibody to Fhit (ZR44, Zymed Laboratories) on formalin-fixed, paraffin-embedded tissues was used. Fhit was constitutively expressed in germ cells, Sertoli cells, and Leydig cells. All 42 cases of intratubular germ cell neoplasia revealed no expression of this protein. No expression of Fhit was observed in any case of pure seminoma or in the seminomatous component of mixed germ cell tumors. Unexpectedly, Fhit expression was frequently (16/18) observed in the glandular tissue of mature teratomatous component of mixed germ cell tumors, despite the absence of Fhit in the intratubular germ cell neoplasia, the presumed precursor lesion. The loss of Fhit expression is a consistent characteristic of intratubular germ cell neoplasia, which suggests a potential role in a maturation/differentiation defect early in the development of testicular germ cell tumors. Likewise, the lack of expression in seminomas is supportive of this view. However, re-expression of Fhit in well-differentiated glandular epithelium of teratomatous component of mixed germ cell tumors suggests that there is no loss of FHIT gene in this subset of neoplasia but rather that Fhit protein expression is differently regulated through the phases of germ cell tumor progression.

The mule (Equus mulus mulus) is a sterile hybrid domestic animal that results from the breeding of a male donkey (Equus asinus) to a female horse (Equus caballus). Usually, spermatogenesis in mules does not advance beyond spermatocytes. In the present study, we performed a comparative and more accurate morphometric and functional investigation of the testis in donkeys and mules. Due to the smaller testis size, lower seminiferous tubule volume density, and fewer germ cells, the total length of seminiferous tubules in mules was significantly smaller than in donkeys. However, the percentage of seminiferous tubules containing germ cells (spermatogonia and spermatocytes) in mules was approximately 95%. The total number of Sertoli cells per testis observed in donkeys and mules was very similar. However, the total number of Leydig cells in mules was approximately 70% lower than in donkeys. At least in part, this difference was probably related to the lower number of germ cells present in mule seminiferous tubules. Although spermatogenesis in mules did not advance beyond secondary spermatocytes/newly formed round spermatids, germ cell associations in the seminiferous epithelium and pachytene spermatocytes nuclear volume in donkeys and mules were similar. The duration of spermatogenesis was estimated using intratesticular injections of tritiated thymidine. Each spermatogenic cycle in donkeys lasted 10.5 days. A similar value was found in mules ( approximately 10.1 days). Considering that the entire spermatogenic process takes approximately 4.5 cycles to be completed, its total duration in donkeys was estimated to last 47.2 days. The results found for mules suggest that the mechanisms involved in the determination of testis structure and function are probably originated from donkeys. Also, the data found for mules suggest that their seminiferous tubules are able to sustain complete spermatogenesis. In this regard, this species is a potential model for transplants of germ cells originated from donkeys and horses or other large animals.

Aim: To determine the effect of oral administration of an aqueous extract from the roots of Lepidium meyenii (maca) on spermatogenesis in adult male rats., Methods: Male rats received an aqueous extract of the root (66.7 mg in one mL) twice a day for 14 consecutive days., Results: Treatment with Lepidium meyenii resulted in an increase in the weights of testis and epididymis but not the seminal vesicle weight. The length and frequency of stages IX-XIV seminiferous tubules, where mitosis occurred, were increased and stages I-VI were reduced in rats treated with Lepidium meyenii., Conclusion: The Lepidium meyenii root invigorates spermatogenesis in male rats by acting on its initial stages (IX-XIV).

The cycle of the seminiferous epithelium in the Java fruit bat, Pteropus vampyrus, and the Japanese lesser horseshoe bat, Rhinolophus cornutus, was investigated by light microscopy and the characteristics of spermiogenesis were compared between these two species. In the Java fruit bat, the cycle of the seminiferous epithelium was divided into 11 stages and developing spermatids were subdivided into 13 steps. While in the Japanese lesser horseshoe bat, the cycle of the seminiferous epithelium was divided into 10 stages and developing spermatids were subdivided into 13 steps. Excepting slight morphological differences, the characteristics of acrosomal formation in both species were almost similar with each other. In the Java fruit bat after stage VII, the acrosome gradually elongated, flattened and finally became scoop-like in shape. In the Japanese lesser horseshoe bat after stage VIII, the acrosome elongated, flattened and then slightly shortened. Before spermiation, the acrosome became long spatula-like in shape. The elongation and flattening of spermatids in these two species were similar to those in insectivores. The finding may reflect the fact that the order Chiroptera is phylogenetically close to the order Insectivora.

During spermatogenesis, germ cells undergo mitotic and meiotic divisions to form haploid round spermatids which mature to functional elongated spermatozoa. During this process there occurs remodeling of cell structure and loss of most of the cytoplasm and a large fraction of cellular proteins. To evaluate the role of the ubiquitin proteolytic system in this protein loss, we measured levels of ubiquitinated proteins and rates of ubiquitin conjugation in extracts of testes from rats of different ages. Endogenous ubiquitin-protein conjugates increased till day 30 and then reached a plateau. In parallel, there was a progressive increase in the rate of conjugation of ubiquitin to proteins in testis extracts from these animals. To test the importance of two major ubiquitin conjugating enzyme families in the conjugation, immunoprecipitation of UBC2 or UBC4 from 10- and 30-day-old testis extracts was carried out and the remaining conjugation activity in supernatants was assayed. Depletion of either enzyme family resulted in decreased conjugation. However, most of the conjugation activity and, more importantly, the increased conjugation during development were UBC4-dependent. Immunocytochemistry demonstrated a marked increase in expression of UBC4 in spermatids, consistent with the UBC4-dependent activation of conjugation seen in vitro. In situ hybridization studies evaluated the contribution of various UBC4 isoforms to this induction. UBC4-1 mRNA was expressed in most cells. UBC4-2 mRNA was restricted to germ cells with high levels of expression in round and elongated spermatids. UBC4-testis had previously been shown to be expressed only in spermatids. Our data suggest that induction of various UBC4 isoforms activates overall conjugation and plays an important role in the cellular remodeling and protein loss occurring during spermatogenesis., (Copyright 1999 Academic Press.)

Testosterone secretion and the expression and relative contents of steroidogenic acute regulatory (StAR) protein and steroidogenic enzymes cholesterol side-chain cleavage cytochrome P450 (P450SCC), 3beta-hydroxysteroid dehydrogenase/delta(5)-->delta(4)-isomerase (3 beta-HSD), and (17)alpha-hydroxylase cytochrome P450/C17-20 lyase (P450(17)alpha) were determined in testicular tissues of bulls treated with a LHRH agonist. Testis morphology and spermatogenesis were also examined. In Experiment 1, bulls (30-mo-old) received no treatment (control, n = 7) or were implanted for 10 days with the LHRH agonist deslorelin (n = 7). Bulls were castrated on Day 10 and testis tissues prepared for Western and Northern blotting. At castration, bulls implanted with deslorelin had greater plasma testosterone (5-fold) and testis content of testosterone (10-fold) compared with control bulls. Relative content (per micrograms total testis protein or RNA) of StAR protein, 3beta-HSD, P450SCC, and mRNA for P450(17)alpha in bulls treated with deslorelin ranged from 3- to 6-fold that of control bulls. In Experiment 2, bulls (20-mo-old) were left untreated (control, n = 6) or implanted with deslorelin (n = 12) for 120 days. On Day 120, bulls were castrated and right testis tissues prepared for morphology. Testis volume and weight were increased (P < 0.01) in bulls treated with deslorelin compared with control bulls. Stereological analysis revealed that this increase occurred in all compartments (seminiferous epithelium, lumen and interstitium) studied, but was significant (P < 0.01) only for the seminiferous epithelium. Absolute numbers of round spermatids per testis were increased (P < 0.05) in bulls treated with deslorelin compared with control bulls. Increased testosterone secretion in bulls treated with deslorelin was associated with increased testicular StAR protein and steroidogenic enzymes. Bulls treated long-term with deslorelin had a faster rate of testis growth and increased daily sperm production at the end of the experiment.

Apolipoprotein J/clusterin/sulfated glycoprotein-2 (apo J) disassociates from spermatozoa and is endocytosed by epithelial cells lining the efferent ducts and epididymis. The low density lipoprotein receptor-related protein-2/megalin (LRP-2) has been shown to bind to apo J and mediates its endocytosis and lysosomal degradation in cultured cells. In this study, immunocytological techniques were used to localize LRP-2 in rat efferent ducts and epididymis and to determine whether its expression correlated with those epithelial cells involved in apo J endocytosis. Pronounced LRP-2 immunochemical staining was observed on the apical surfaces of epithelial cells lining the efferent ducts and in the intermediate zone, proximal caput, and corpus and cauda regions of the epididymis. Single immunogold labeling at the electron microscopic level showed LRP-2 to be present within coated pits, endocytic vesicles, and early endosomes of the nonciliated cells of the efferent ducts and the principal cells of the epididymis. In efferent ducts, double immunogold labeling showed both LRP-2 and apo J to be present in endocytic compartments including coated pits, endocytic vesicles, and early endosomes of nonciliated cells. However, while apo J was detected in late endosomes and lysosomes of nonciliated cells, LRP-2 was not. Apical tubules, possibly emerging from late endosomes, contained labeling for LRP-2 but not for apo J. Ciliated cells lying adjacent to nonciliated cells displayed no labeling for either LRP-2 or apo J. These results are consistent with the possibility that LRP-2 serves as an endocytic receptor for apo J in vivo and that after endocytosis the LRP-2 is recycled back to the cell surface while apo J is delivered to the lysosomes for degradation. To provide additional evidence implicating LRP-2 in apo J endocytosis, a receptor-associated protein (RAP), an antagonist of apo J binding to LRP-2, was injected into the efferent duct lumen. Subsequent immunocytological analysis of the efferent duct showed that the RAP treatment abolished the endocytosis of apo J by the nonciliated cells. Taken together, these data indicate that LRP-2 is a likely mediator of apo J endocytosis by the nonciliated efferent duct cells.

We have performed an experimental study on rats and dogs to evaluate the long term effects (from 1 to 12 months) of vasectomy on the structure of the testis. From four months after vasectomy onwards, the specimens showed very important changes in the seminiferous epithelium and Sertoli cells, with an obvious thickening of the basement membrane that supports the epithelium. The deterioration depended on the time passed and, over six months after vasectomy, the alterations were very clear and the seminiferous tubules became atrophic and shrunk, sometimes without any remains of seminiferous epithelium and with an important hypertrophy of the interlobular interstitial tissue, although we did not see an increase in the number of Leydig cells. Alterations due to vasectomy depend on the animal species, the peculiarities of techniques and, of course, the time passed after surgery.

The frequencies of the cellular associations of the seminiferous epithelium were determined at various ages after birth in immature Djungarian hamsters and Wistar rats. The frequencies of the cellular associations present in immature animals were then compared with the frequencies of the corresponding pooled stages in adult animals. At 15 days of age, the cellular associations present in Djungarian hamsters could be divided into four groups based on the presence of A3, intermediate (In), or B (B1 and B2) spermatogonia, or preleptotene-leptotene-zygotene spermatocytes. Compared with adult animals, the group containing spermatocytes was found to be enriched by 52%, while the frequency of the B spermatogonia was reduced by 55%. At 22 days of age, the cellular associations of spermatogenesis were classified into five groups: pachytene spermatocytes associated with A3 spermatogonia. In spermatogonia, B spermatogonia, or preleptotene-leptotene-zygotene spermatocytes, or the meiotic divisions. The stages containing preleptotene-leptotene-zygotene spermatocytes and those containing A3 spermatogonia were enriched by 35% and 59% compared with adult values, respectively. The meiotic divisions were enriched by 21%, but this increase was not significant. The frequency of the B spermatogonia was reduced by 62%. In addition, the synchronization factor, calculated with the computer program SYNTEST, was increased to 1.48 +/- 0.05 (mean +/- SEM) and 1.38 +/- 0.03 at 15 and 22 days of age, respectively, and was significantly higher (P < 0.001 at both ages) than the adult value of 1.08 +/- 0.02. However, after 22 days of age the synchronization factor did not differ from that in adult animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Stages of the spermatogenic cycle in human seminiferous tubules were evaluated in men with varied efficiencies of spermatogenesis to determine if the architectural arrangement of stages or the atypical cell types contributed to variation in sperm production rates. Testes were selected from men with low, intermediate, and high daily sperm production per g parenchyma (DSP/g). Round tubular cross sections were photographed by bright-field microscopy. Stages were identified for each cross section by two observers and the number of stages represented in each cross section was recorded. Number of stages per cross section in men with low efficiency of spermatogenesis were significantly (P < 0.05) fewer than men with intermediate and high efficiency of spermatogenesis. Further, the percentage of stages with atypical cell types in men with high DSP/g was significantly (P < 0.05) higher than men with low DSP/g. There was a significant relationship (P < 0.01) between the percentages of stages with atypical cell types per stage and number of stages per cross section. The atypical cell types appear to result from high density of stages per cross section in men with high DSP/g. There was no significant difference observed between groups for tubular volume, diameter, length, volume density, and volume density of seminiferous epithelium. However, a significant (P < 0.05) positive correlation between percent seminiferous epithelium per testis with DSP/g or with the number of stages per cross section was found.(ABSTRACT TRUNCATED AT 250 WORDS)

Testis size, seminiferous tubules and plasma testosterone concentrations showed conspicuous annual changes in Sika deer of Hokkaido, Japan. The onset of the spermatogenic process occurred in July or August. Spermatogenic activity had already reached its height in late October, at the beginning of the rutting season, and had begun to decline in late December. Spermatogenesis had stopped in February or March. Plasma testosterone concentrations showed very high levels in late October and early November, but was almost at the basal level in February, March, June and December. The wide individual variation of the plasma levels in October suggest pulsatile secretions of testosterone.

The duration of the cycle of the seminiferous epithelium of the South American white-belly opossum, as obtained by autoradiography after intratesticular injection of tritiated thymidine, was estimated to be 17.3 +/- 0.1 days (mean +/- s.d.). Quantitative histological analysis was performed on testes from animals caught both in mating and non-mating periods of the annual reproductive cycle. Significant differences were found in the volumetric proportion of Leydig cells, but the spermatogenic yield remained constant throughout the year. The numerical ratio between type A spermatogonia and zygotene primary spermatocytes (1:12.0), as well as daily sperm production (4.8 x 10(6) sperm cells per g of testis parenchyma per day), were found to be lower than those reported in most eutherian mammals.

To probe the relationship between the size of the Sertoli cell population, established during perinatal development, and production of germ cells in the adult testis, a Sertoli cell-depleted rat model was developed. This was accomplished by delivering an antimitotic drug, cytosine arabinoside (araC), directly to the testis of newborn pups. Initial studies of these araC-treated neonates indicated that 1) the drug is cleared rapidly from the testis; 2) it substantially reduces the level of Sertoli cell proliferation; 3) Sertoli cell division ceases at a normal time in spite of the previous drug treatment; and 4) araC itself has no residual effect on germ cell proliferation, which begins several days after the injection. Pups given araC were allowed to reach maturity, and their testes were perfuse-fixed for light microscopic morphometry. When the numbers of Sertoli cells in adult rats given araC as were compared with those in normal littermates, a 54% decrease in the size of the Sertoli cell population was detected in treated rats, now referred to as Sertoli cell-depleted. Moreover, when round spermatids were quantified and compared in normal and Sertoli cell-depleted adults, testes of the latter were found to contain 55% fewer round spermatids. Since, in the araC-treated group, the decrease in Sertoli cell population size was paralleled by a reduction in spermatid production of equal magnitude, the number of round spermatids per Sertoli cell was essentially identical in normal and Sertoli cell-depleted animals. Measurements of serum androgen-binding protein (ABP) and FSH in both groups indicated that the circulating level of ABP in Sertoli cell-depleted rats was approximately half, and the concentration of FSH approximately twice, that in normal animals. Thus, even though FSH is elevated in Sertoli cell-depleted rats, the production of ABP per Sertoli cell is unchanged. In addition, collective volume of Leydig cells and ventral prostate weights were normal in the Sertoli cell-depleted group, suggesting that Leydig cell function in these rats is normal. In summary, a Sertoli cell-depleted rat model has been produced by interfering specifically with Sertoli cell proliferation early in postnatal life, before onset of germ cell division. Moreover, our findings with this model indicate that production of normal numbers of germ cells in adults depends, at least in part, on the size of the Sertoli cell population. Thus, our observations identify the perinatal period, when the Sertoli cell population is established, as critical for development of quantitatively normal spermatogenesis in the adult.

Spermatogenesis in the non-scrotal, insectivorous microchiropteran, Rhinopoma kinneari Wroughton, is a cyclic phenomenon. It comprises (i) eleven cell associations, and (ii) 16 evolutive steps in spermiogenesis. In the seminiferous epithelium, spermatogonia of the A, intermediate (In), and B type are characteristically present. Leptotene, pachytene and diakinesis stages were easily discerned in dividing spermatocytes. During the 16-step spermiogenesis, the acrosomic system develops through the Golgi, cap, acrosome and maturation phases. The relative frequency of various cell associations differs markedly and is significantly at variance with other mammalian species. Fully formed spermatozoa are dimorphic with conical or blunt heads.

The Sertoli cells of the Cape horseshoe bat (Rhinolophus capensis) and Schreiber's long-fingered bat (Miniopterus schreibersii) undergo marked changes in ultrastructure related to stages in the spermatogenic cycle. The amount of lipid stored in the Sertoli cells varies annually and is at a maximum from just after spermiation to early in the following spermatogenic cycle. During spermatogenesis, the diameter of the lipid droplets decreases, reaching a minimum prior to spermiation. Sertoli cells exhibit a marked apicobasal differentiation, particularly in the vicinity of developing late spermatids, where the cytoplasm of the Sertoli cell is packed with smooth endoplasmic reticulum. The possible roles of lipid droplets and smooth endoplasmic reticulum. The possible roles of lipid droplets and smooth endoplasmic reticulum in steroidogenesis by Sertoli cells are discussed. Junctional complexes occur between Sertoli cells and spermatogonia, are apparently absent from between Sertoli cells and spermatocytes, and are restricted to the region of the developing acrosome in the spermatids. Annulate lamellae, which occur commonly in the developing germinal cells and less frequently in the Sertoli cells, may be associated with the production of microtubules, which are present in both spermatids and Sertoli cells.

For elucidating the problem whether the tissue composition of the male gonad is identical or different in the equatorial zone and polar zones of the organ, 104 left testes were studied obtaining them from the cadaver of patients dying from various acute and chronic diseases not affecting the endocrine system. In the histological preparations of the polar and equatorial zones of the testis, the proportion [%] were calculated of 6 types of convoluted tubules designated with numbers from 1 to 6 in the sequence of decreasing spermatogenic activity, and by the morphometric method, the proportions [%] were determined of the tubular lumina, spermatogenic epithelium, tubular membrane, stroma, blood vessels, and LEYDIG's cells. In the whole analysed group, only tubules type 2 were more numerous in the equatorial part, while tubules type 6 prevailed in the polar parts. The lumina of the tubules occupied a greater % of the testicular structure in the polar parts as compared with the central part. Other tissue elements were present in similar amounts or the differences between their amounts in these parts were only nearly significant statistically. The study showed also that the number of type 1 tubules decreased significantly with progressing age, while the number of type 6 tubules increased. A statistically significant fall of the proportion [%] of the spermatogenic epithelium and increased proportion of tubular membranes and stroma were another age-related changes. It is worth stressing that these age-related processes progressed parallelly in the equatorial part and in the polar parts of the testis which suggest that ageing of the organ occurs simultaneously in the whole gonad.

With the use of a digital image-processing method three-dimensional reconstructions of the arrangement of spermatocytes in human seminiferous tubules were performed. With this method it was possible to investigate the cellular distribution in the tubule in nearly any given perspective and projection. In addition, by means of simple mathematical procedures, such as by transformation of Cartesian coordinates into cylindrical coordinates, it was possible to vary the shape of a reconstruction, i.e., to convert the cylindrical image of a tubular portion into a right-angled r-phi-z-representation. The present work not only confirms the existence of a complex helical plan of organization of the human seminiferous epithelium but also provides further aspects of the phenomenon of physiological germ-cell loss and its integration into the kinetics of spermatogenesis.

Testicular tissue and sperm from the cauda epididymidis of 300 boars were obtained in 6 collections during 1 year at a Chicago abattoir. Seventy-five boars (25%) had - 1 sperm abnormality present at rates that could result in infertility. These abnormalities and their frequency criteria were: proximal droplets > or = 15%, loose heads > or = 10%, abnormal middle pieces > or = 10%, abnormal acrosomes > or = 10%, bent tails > or = 15%, coiled tails > or = 15%, and abnormal head shapes > 5%. These 75 abnormal boars were then compared with a control group of 75 boars selected at random from the remaining 225 boars. Frequency of sperm abnormalities, testicular and epididymal weights, relative seminiferous epithelial areas (SEA), and the degree of testicular lesions were used in making comparisons. Testicular lesions were scored on a scale of 1 (normal) to 5. The SEA was measured and was expressed as the mean area of 50 tubules. The SEA was negatively correlated with proximal droplets and abnormal middle pieces in abnormal boars and abnormal head shapes in control boars. The mean SEA was 8.71 and 5.60 cm2 for control and abnormal boars, respectively. The overall mean score for testicular lesions was 1.72 and 3.17 for control and abnormal boars, respectively.

The present study was carried out to determine the detailed histological and cytological features of the excurrent ducts of the male reproductive system in the rabbit. The rete testis is comprised of flattened epithelium surrounding a wide lumen which is devoid of spermatozoa. Vasaefferentia epithelium is comprised mainly of ciliated and non-ciliated cells and the lumen is devoid of sperms. On the basis of the characteristics of the epithelium, and luminal contents and histometric observations such as luminal diameter, epithelial cell height and stereocilia height, seven regions of epididymis were identified. Similarly, the vas deferens was identified into 3 regions; proximal, distal and terminal. Results were discussed bearing in mind the functional significance of different regions.

The present work reports a tridimensional reconstruction of the intratesticular seminal pathway in the hamster as studied with light microscopy of serial sections of the proximal pole of the testis. It has three portions: a "terminal segment" of the seminiferous tubules; the tubuli recti with three parts; and the rete testis with two parts, intratesticular and extratesticular. Experiments using ductuli efferentes ligation show a marked dilation of the rete testis and tubuli recti without a corresponding dilation of the seminiferous tubules. This may be due to a valve-like action of the cells in the terminal segment of the tubules.

Testes from 8 adult rats were perfusion-fixed with buffered glutaraldehyde and semi-thin sections of the seminiferous epithelium at all stages of the spermatogenic cycle were subjected to morphometric analysis at the light microscope level. Methods are presented to derive the volume occupied by the Sertoli cells within the total volume of the seminiferous epithelium at each stage of the cycle for a single testis. The total number of Sertoli cells present in each stage for the whole testis was calculated from a measurement of Sertoli cell numerical density and the total volume of the seminiferous tubule in the testis at each stage of the cycle. Average volume of a single Sertoli cell for each stage was derived by dividing the first set of data by the latter data. Average Sertoli cell volume exhibited a cyclic variation in relation to the stages of the spermatogenic cycle. Sertoli cells were smallest during stages VI-VIII (5300-5500 micron3) increased to maximum volume during stages XII-XIV (7700-8000 micron3) and thereafter during stages I-V, gradually contracted in volume to complete the cycle. Stage-dependent cyclic variations in Sertoli cell volume offers evidence that the morphology of the Sertoli cell undergoes structural modifications to accommodate changes in the shape and volume of the developing germ cells. Furthermore these volume changes implicate the Sertoli cells in cyclic metabolic, absorptive and secretory functions which possibly direct the maturation of germ cells during the spermatogenic cycle.

To date, there are no comprehensive quantitative studies detailing the effect of vasectomy on the mouse testes. We examined the effects of vasectomy on testicular germ cell numbers using detailed light microscopy, and the effect of long-term vasectomy on sperm transport to the caput epididymis using radiolabelled germ cells. Short-term vasectomy has no significant effects on the number of germ cells within the testes; however, after long-term vasectomy, spermiogenesis was affected. Surprisingly, even after long-term vasectomy, the rate of spermatogenesis and sperm transport to the epididymis was unaffected.

Tissue sections from testes and epididymides obtained from 17 young beef bulls with scrotal circumference (SC) between 27 and 40.5 cm were studied to determine whether small testes were a manifestation of lesions or a result of less, but otherwise normal, seminiferous epithelium. The SC correlated negatively with the estimates of germinal epithelial loss and positively with seminiferous epithelial area. Four bulls with SC less than 30 cm had severe lesions in their testes. Hypoplastic tubules were characterized by Sertoli's cells only with no evidence of germinal cells. Loss of germinal cells, leaving vacuolated epithelium and atrophy, were observed in degenerated tubules. Hyperplasia of Leydig's cells was observed in the vicinity of Sertoli's cell-only tubules, resulting either from degeneration or hypoplasia, and atrophy of Leydig's cells was associated with tubules devoid of Sertoli's cells. These findings indicated that Sertoli's cells may produce a factor(s) required for maintenance and regulation of Leydig's cell function. Epididymal epithelium, especially in the head, had regressed in bulls with hypoplastic and degenerative changes in their testes. Decreased sperm concentration and motility and an increased frequency of morphologic defects were observed in the 4 bulls with testicular lesions and regressed epididymal epithelium. Blood plasma profiles of cortisol, follicle-stimulating hormone, luteinizing hormone, and testosterone were determined in the 4 bulls with SC less than 30 cm and 10 of the 13 bulls with SC greater than 30 cm. There were no statistically significant (P greater than 0.1) differences in the responses to exogenous gonadotropin-releasing hormone or base-line patterns of blood plasma follicle-stimulating hormone and luteinizing hormone between the 2 groups. However, in the bulls with SC less than 30 cm, the mean concentration of testosterone was lower, whether spontaneous (P less than 0.05) or exogenous gonadotropin-releasing hormone induced (P less than 0.1). The fact that these bulls were not deficient in gonadotropins indicated that Leydig's cell function was impaired by local factors, either the factors that caused the tubular damage or those consequent to the tubular damage.