Structural, ultrastructural and immunohistochemical evidence of testosterone effects and its ablation on the bulbourethal gland of the Artibeus planirostris bat (Chiroptera, Mammalia).

This study evaluated the effects of testosterone in the bulbourethral glands (BG) of the bat, Artibeus planirostris, by performing castration and posterior hormonal supplementation of the animals. The results showed a decrease in testosterone levels in animals 15days after castration, which induced a small reduction in epithelium height, percentage of AR+ cells, and an increase in the amount of basal cells. This reduction became more severe in groups castrated for longer periods (19 and 22days), where there was also an increase in apoptotic cells. Moreover, the hormonal supplementation increased testosterone levels (after 3 and 7days of supplementation), causing a glandular reactivation that increased the epithelium height and AR expression. In conclusion, BG took longer to respond to ablation of testosterone than other reproductive glands, since it showed evident aspects of regression only in animals 22days after castrated.

Comparative analysis of the male reproductive accessory glands of bats Noctilio albiventris (Noctilionidae) and Rhynchonycteris naso (Emballonuridae).

In eutherian mammals, the male reproductive accessory glands (RAGs) comprise the prostate, bulbourethral glands, ampullary glands, and the seminal vesicles. Their composition, anatomy and function vary widely between species. This study aimed to characterize histologically and compare the RAGs of bats. The RAGs of Noctilio albiventris (Noctilionidae) and Rhynchonycteris naso (Emballonuridae) were studied using anatomical and histological methods, and were reconstructed three dimensionally. The RAGs of N. albiventris and R. naso are composed of a compact glandular complex that surrounds the urethra and a pair of bulbourethral glands, which are extra-abdominally located in the inguinal region. In both species, the glandular complex is composed of two well-defined prostatic regions (ventral and dorsal). The ventral region showed an atypical epithelium (holocrine), where no obvious cellular limits were observed, and PAS-positive secretion. The dorsal region had a pseudostratified cuboidal epithelium, with basal and secretory cells, and PAS-negative secretion. Noctilio albiventris also had urethral glands (Littre glands) surrounding the urethra, however, R. naso had only muscles. Both species had bulbourethral glands, with simple columnar epithelium and PAS-positive secretion. In conclusion, the RAGs of N. albiventris and R. naso comprised a pair of bulbourethral glands and an intra-abdominal complex, composed of a prostate with two different regions (ventral and dorsal), while the ampullary glands and seminal vesicles were missing in both species. This morphology was more closely related between N. albiventris and R. naso, and to species of the family Phyllostomidae than to families Molossidae and Vespertilionidae. J. Morphol. 277:1459-1468, 2016. © 2016 Wiley Periodicals, Inc.

Morphophysiology and ultrastructure of the male reproductive accessory glands of the bats Carollia perspicillata, Glossophaga soricina and Phyllostomus discolor (Chiroptera: Phyllostomidae).

The male reproductive accessory glands (RAGs) are important organs that contribute to the secretion of different substances that composed the ejaculate. Despite this important function, their composition, anatomy and function vary widely between species. Thus, the RAGs of three species of phyllostomid bats were morphologically and ultrastructurally characterized and compared in this study. The RAGs of the three analyzed species are composed of a prostate and a pair of bulbourethral glands (BG). In all species, the prostate is composed of three well-defined regions (ventral, dorsolateral and dorsal regions). The ventral region showed an atypical epithelium (undefined) with no obvious cellular limits and a holocrine PAS-positive secretion. The dorsolateral region of Carollia perspicillata and Phyllostomus discolor showed a pseudostratified cubic morphology, and that from Glossophaga soricina had a columnar morphology endowed with cytoplasmic projections and stereocilia. The dorsal region of the three analyzed species is composed of a pseudostratified columnar epithelium endowed with stereocilia; however, G. soricina also presented cytoplasmic projections in the apical portions of the secretory cells similar to those in the dorsolateral region. The BG of the three analyzed species are composed of a pseudostratified columnar epithelium including basal and PAS-positive secretory cells. In conclusion, this study morphologically and ultrastructurally characterized the RAGs of three species of phyllostomid bats, demonstrating the presence of a novel third prostatic region in species of this family. The results also showed the absence of seminal vesicles and ampullary glands, and better characterized the holocrine pattern of the prostatic ventral region, which is unique to bats.

The effects of castration followed testosterone supplementation in prostatic complex of Artibeus planirostris (Chiroptera: Phyllostomidae).

The prostatic complex (ventral and dorsal regions) of Artibeus planirostris exhibits seasonal variations throughout the year. Circulating testosterone was correlated with prostate weight, showing an increase from autumn to summer, with the highest peak in summer corresponding to the largest breeding season. This indicates that the level of serum testosterone influences variations in both testicular and prostatic weights. Serum testosterone levels seem to be closely related to the different responses of these glands throughout the year. The castration (consequent suppression of testosterone) and subsequent hormone supplementation may elucidate the relationship of these two glandular types with testosterone. Thus, the aim of this study was to evaluate the effect of castration and the testosterone supplementation in the male prostatic complex of A. planirostris. The results indicated that both prostatic regions were affected by the ablation of testosterone, presenting a decrease in cell proliferation and an increase in apoptosis. Similarly, the prostate was responsive to hormonal supplementation, having a recovery of the active morphophysiological pattern with testosterone supplementation. However, data have shown that the ventral region was more sensitive to changes in testosterone than the dorsal, presenting greater cell renewal.

Comparative analysis of the male reproductive accessory glands of bat species from the five Brazilian subfamilies of the family Phyllostomidae (Chiroptera).

This study aimed to morphologically characterize and compare the male reproductive accessory glands (RAGs) of bats belonging to the five Brazilian subfamilies of the family Phyllostomidae (Carollia perspicillata-Carollinae; Desmodus rotundus-Desmodontinae; Glossophaga soricina-Glossophaginae; Phyllostomus discolor-Phyllostominae and Platyrrhinus lineatus-Stenodermatinae). The study demonstrated that the RAGs of phyllostomid bats were comprised of a pair of extra-abdominal bulbourethral glands and an intra-abdominal complex, composed of paraurethral glands and a prostate with two (Desmodontinae and Stenodermatinae) or three (Carollinae, Glossophaginae and Phyllostominae) different regions, with the absence of the seminal vesicles; this pattern possibly evolved from a process of compaction of the prostatic regions from an ancestor with three regions.

Structure, histochemistry and seasonal variations of the male reproductive accessory glands in the Pallas's mastiff bat, Molossus molossus (Chiroptera: Molossidae).

Due to their wide geographical distribution, bats suffer considerable influence from abiotic factors on their reproductive strategies, detected through behavioural or functional assessment of the gonads and accessory glands. The present study aimed to characterise anatomically and morphologically the reproductive accessory glands (RAGs) of Molossus molossus (Molossidae) and evaluate their seasonal variations. The RAGs were removed, fixed, sectioned after histological processing and submitted to the following stains: haematoxylin-eosin, periodic acid--Schiff (PAS) and Gömöri's reticulin. Our data demonstrated that the RAGs of M. molossus are composed of a semi-lobed prostatic complex associated with the urethra and a pair of inguinal bulbourethral glands. Histology and three-dimensional reconstruction of the prostatic complex demonstrated the existence of two regions: ventral and dorsal (named according to the position around the urethra). The two regions had distinctive morphological and histological differences, with the ventral region being the most prominent and the dorsal formed by four lobes. Considering the seasonal evaluation, we can infer that the prostatic complex of M. molossus is active throughout the seasons in São Paulo State (Brazil) and, although each of the prostatic regions has inherent characteristics, they synchronise to establish the main reproductive peak in spring (early rainy season).

Structure, histochemistry, ultrastructure and seasonal variations of the male prostatic complex in the black Myotis bat, Myotis nigricans (Chiroptera: Vespertilionidae).

Chiroptera are one of the most diverse orders of mammals and a unique group within Mammalia that posses a wide geographic distribution and considerable variability in reproductive strategies. The aims of the present study were to characterise the male prostatic complex of the bat Myotis nigricans (Vespertilionidae) and evaluate seasonal variations in the prostatic complex of M. nigricans specifically. Twenty-three sexually mature specimens (four sample groups: winter, spring, summer and autumn) were subjected to macroscopic, microscopic, morphometric and ultrastructural analyses. The reproductive accessory glands of M. nigricans were found to be composed of a multilobed complex associated with the urethra and a pair of inguinal bulbourethral glands. The complex was composed of three bilobed prostatic regions (ventral, dorsolateral and dorsal) with no ampullary gland and seminal vesicles. This pattern of lobulation is very similar to that described for the prostate of rodents; however, it differs from that of other mammals and even other families of bats (e.g. Phyllostomidae and Molossidae). Each prostatic region in M. nigricans has unique and distinctive characteristics, which synchronise to establish the main reproductive peak of the species in summer. The data also indicated an asynchrony in the activity of primary and secondary reproductive organs in the annual reproductive cycle of M. nigricans in São Paulo State, Brazil.

Seasonal changes in the prostatic complex of Artibeus planirostris (Chiroptera: Phyllostomidae).

The male reproductive accessory glands are important organs that secrete products that ensure the survival, viability and motility of spermatozoa, not only in the male reproductive tract, but also in the female. Most studies relating to the reproduction of bats do not include these glands, and detailed studies of these glands describing annual variations in the morphology and physiology are scarce. The aim of this study was to evaluate the impact of these variations on morphophysiology of the prostatic complex (PC) of Artibeus planirostris (Chiroptera: Phyllostomidae). Adult males were collected from June 2009 to July 2010, and the PC was subjected to various analyses. We observed that the PC showed marked variations throughout the year. Both PC and testicular weight increased synchronously with an increasing rate of circulating testosterone, from the autumn until summer, demonstrating that both organs are regulated by this hormone. Each region of the PC (ventral and dorsal) was unique and distinct. The ventral region showed the glandular lumen as a predominant component, in contrast to the dorsal introduced epithelium; acid phosphatase activity was observed in the epithelium of all acini in the dorsal region, but only in the stroma of the ventral region, in addition each region responded differently to variations in the environment and circulating testosterone. We concluded that the PC of A. planirostris produces two secretory peaks, which were related to a gradual increase in the rate of circulating testosterone, which stimulates both of the prostate regions to prepare for the two reproductive periods.

Annual reproductive cycle of males of the flat-faced fruit-eating bat, Artibeus planirostris (Chiroptera: Phyllostomidae).

Artibeus planirostris is an endemic species of Phyllostomid bat from the Neotropical region. Some studies have indicated that it exhibits seasonal bimodal polyestry; however, others postulate that it may be able to produce young at any time during the year. Thus, the aim of this study was to evaluate the annual variations in testicular and epididymal parameters of this species in southeast Brazil and try to understand how the reproduction of this species is regulated in this environment. Sixty mature male specimens, collected between June 2009 and May 2010, were submitted to morphometric and immunohistochemical analysis. Our study showed that A. planirostris presented a continuously active pattern of spermatogenesis throughout the year, presenting spermatozoa inside its cauda epididymis in all months, but with two pronounced peaks of spermatogenic production, one in September and other in February. We propose that the males developed these two peaks in order to produce sufficient sperm for the reproduction in a harem system and to synchronize with the female reproductive cycle, which had a bimodal polyestric pattern. Control of this variation is directly linked to the expression of the androgen receptor (AR) in Sertoli cells and to serum testosterone levels, which appear to synchronize to establish these two peaks. In the months preceding the two peaks, the testis have a higher expression of the AR, which possibly stimulates the increase in PCNA, and drives a gradual increase in the testicular parameters. Taken together the results suggest that if sperm storage happens in this species, it is of short duration.

Structure, histochemistry and ultrastructure of the male reproductive accessory glands in the neotropical flat-faced fruit-eating bat Artibeus planirostris (Chiroptera: Phyllostomidae).

Chiroptera, the second largest mammalian order, presents different reproductive strategies and unique reproductive features. However, there are few reports regarding male reproductive accessory glands (RAGs) in Chiroptera. Thus, the aim of the present study was to characterise the RAGs of the exclusively neotropical bat Artibeus planirostris (Chiroptera: Phyllostomidae) macroscopically, microscopically and ultrastructurally. The RAGs were composed of a prostatic complex with two regions (ventral and dorsal) and paraurethral and bulbourethral glands, but no seminal vesicles. The ventral region had an undefined epithelium, with secretory and basal cells, and its secretions were periodic acid-Schiff (PAS) positive. The dorsal region received both deferens ducts, had a columnar pseudostratified epithelium with secretory and basal cells. There were two types of secretions from the dorsal region: one that was basophilic and another that was mixed PAS positive and PAS negative. The paraurethral glands were dispersed in the connective tissue of the urethra, whereas the bulbourethral glands were located in the penile root. Histological and ultrastructural data confirmed the prostatic nature of the ventral and dorsal regions and the holocrine nature of the ventral region, with the latter finding never having been described previously for the prostate gland. Our findings demonstrate the wide discrepancy of RAGs between A. planirostris and other mammals in terms of their composition, structure and morphology.

Morphological variation of primary reproductive structures in males of five families of neotropical bats.

Bats present unique features among mammals with respect to reproduction, and although neotropical bats do not have a hibernation period, many of their reproductive characteristics vary seasonally and interspecifically. Thus, this work aimed to examine the reproductive structures of 18 species belonging to five families of Brazilian bats. The gross anatomy of the testes varied little; however, the epididymis of Emballonuridae and Vespertilionidae showed exceptional structures with a large elongation of the caudal region. We observed a wide variation in the positioning of the testes: Phyllostomidae and Noctilionidae presented external testes; Emballonuridae and Molossidae presented migratory testes that may be in intra-abdominal or external positions; and Vespertilionidae displayed scrotal testes. In the histological evaluation, we observed a different pattern in vespertilionid species, with testicular regression and sperm retention/storage in the cauda epididymis in the five species analyzed. Similar testicular regression was observed in Molossops temminckii; however, sperm retention/storage was not observed in this species. These data suggest that although the species analyzed are tropical species that do not present a prolonged period of torpor (hibernation), they still maintain a period of seminiferous tubule regression and sperm storage very similar to that observed in hibernating bats.

Ultrastructure of spermatogenesis in the white-lined broad-nosed bat, Platyrrhinus lineatus (Chiroptera: Phyllostomidae).

Spermatogenesis, the remarkable process of morphological and biochemical transformation and cell division of diploid stem cells into haploid elongated spermatozoa, is one of the most complex cell differentiations found in animals. This differentiation process has attracted extensive studies, not only because the process involves many radical changes in the cell shape and biochemistry, but also because the phases and steps of differentiation have provided a better basis for analyzing the seminiferous epithelium cycle. Thus, this study aimed to characterize ultrastructurally the spermatogenesis process in the bat Platyrrhinus lineatus in order to provide a basis for determining the stages of spermatogenesis and to facilitate comparisons of the process between bat species and other vertebrates. Based on ultrastructural characteristics three main types of spermatogonia could be accurately identified: A(d), A(p) and B; the differentiation of spermatids was clearly divided into 12 steps (steps 1-3: Golgi phase, steps 4-5: cap phase, steps 6-9: acrosomal phase and steps 10-12: maturation phase). The ultrastructure of spermatozoa, Leydig cells and Sertoli cells was characterized; and some processes including nucleolar disorganization and the formation of synaptonemal complexes, acrosome and chromatoid body were discussed. Based on our results we may conclude that the spermatogenic process of P. lineatus follows the pattern of mammals with some specificity, as the process of formation of the acrosome and the presence of the perfuratorium. By other side, the simpler ultrastructure of its spermatozoon shows a pattern more closely related to the sperm cells of humans and other primates.