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.

Seasonal morphological variations and age-related changes of the seminal vesicle of viscacha (Lagostomus maximus maximus): an ultrastructural and immunohistochemical study.

The viscacha is a seasonal rodent that exhibit an annual reproductive cycle with periods of maximum reproductive activity and gonadal regression. We studied seasonal variations in the morphology and cellular population of the seminal vesicles (SVs) during both periods and in impuber animals. Seminal vesicles were studied by light and electronic microscopy. Measurements of epithelial height, nuclear diameter, luminal diameter, and muscular layer were performed. Also, we studied the distribution of androgen receptors (AR) in this gland during the reproductive cycle and in impuber animal. During gonadal regression, principal and clear cells showed signs of reduced functional activity. These were characterized by an epithelium of smaller height, irregular nuclei, and cytoplasm with few organelles, dilated cisterns, and glycogen granules. In impuber animals, the principal cells showed large nuclei with chromatin lax and cytoplasm with small mitochondria, poorly developed Golgi apparatus, and granules of glycogen. On the other hand, the cells exhibited seasonal variations in the distribution and percentage of immunolabeled cells to AR throughout the annual reproductive cycle. During the gonadal regression period, glandular mucosa exhibited numerous epithelial cells with intense nuclear staining. However, fibromuscular stromal cells were weakly positive for AR in contrast to what was observed during the activity period. Considering that testosterone values are lower in adult animals during the period of gonadal regression and in impuber animals, our immunohistochemical results show a significant correlation with the percentage of AR-immunopositive cells. In conclusion, these results demonstrate that the structure of the SVs changes in the activity period of viscacha, probably because of elevated levels of testosterone leading to an increase in the secretory activity of epithelial cells.

Effect of chronic treatment with Rosiglitazone on Leydig cell steroidogenesis in rats: in vivo and ex vivo studies.

BACKGROUND: The present study was designed to examine the effect of chronic treatment with rosiglitazone - thiazolidinedione used in the treatment of type 2 diabetes mellitus for its insulin sensitizing effects - on the Leydig cell steroidogenic capacity and expression of the steroidogenic acute regulatory protein (StAR) and cholesterol side-chain cleavage enzyme (P450scc) in normal adult rats. METHODS: Twelve adult male Wistar rats were treated with rosiglitazone (5 mg/kg) administered by gavage for 15 days. Twelve control animals were treated with the vehicle. The ability of rosiglitazone to directly affect the production of testosterone by Leydig cells ex vivo was evaluated using isolated Leydig cells from rosiglitazone-treated rats. Testosterone production was induced either by activators of the cAMP/PKA pathway (hCG and dbcAMP) or substrates of steroidogenesis [22(R)-hydroxy-cholesterol (22(R)-OH-C), which is a substrate for the P450scc enzyme, and pregnenolone, which is the product of the P450scc-catalyzed step]. Testosterone in plasma and in incubation medium was measured by radioimmunoassay. The StAR and P450scc expression was detected by immunocytochemistry. RESULTS: The levels of total circulating testosterone were not altered by rosiglitazone treatment. A decrease in basal or induced testosterone production occurred in the Leydig cells of rosiglitazone-treated rats. The ultrastructural and immunocytochemical analysis of Leydig cells from rosiglitazone-treated rats revealed cells with characteristics of increased activity as well as increased StAR and P450scc expression, which are key proteins in androgen biosynthesis. However, a number of rosiglitazone-treated cells exhibited significant mitochondrial damage. CONCLUSION: The results revealed that the Leydig cells from rosiglitazone-treated rats showed significant reduction in testosterone production under basal, hCG/dbcAMP- or 22 (R)-OH-C/pregnenolone-induced conditions, although increased labeling of StAR and P450scc was detected in these cells by immunocytochemistry. The ultrastructural study suggested that the lower levels of testosterone produced by these cells could be due to mitochondrial damage induced by rosiglitazone.

Evaluation of the ethanol intake on the Calomys callosus seminal vesicle structure.

The effects of chronic alcohol ingestion on the structure of the glandular epithelium of the seminal vesicle of the rodent Calomys callosus were analyzed in 24 adult animals aged 3 months divided into three experimental groups. The control group received a solid diet and tap water, the alcoholic group received the same solid diet and ethanol P.A. diluted 20% in water (v/v) for 4 months. The abstinent group received the same liquid diet of the alcoholic one for the same period and after that the alcoholic diet was changed by water for a period of 3 months. After treatment, all animals were anesthetized, weighed and sacrificed. At the end of treatment, mean body weight did not differ between animal groups. The glandular epithelial cells of the alcoholic and abstinent groups showed atrophy and ultrastructural alterations such as the presence of altered nuclei, intense dilatation of the cisterns of the granular endoplasmic reticulum, intense digestive vacuoles and lipid droplets. Ethanol ingestion provokes marked lesions on the epithelium of the seminal vesicle probably interfering on the glandular secretion.

Differential proliferative response of the ventral prostate and seminal vesicle to testosterone replacement.

We have investigated epithelial cell proliferation and the rate of glandular recovery of the ventral prostate (VP) and seminal vesicle (SV) promoted by testosterone replacement (TR) in castration-induced regressed glands. Adult male Wistar rats were castrated and, after 21 days, they were treated with testosterone propionate (4 mg/kg/day). Intact (CT) and castrated rats without TR (CS) were also analysed. VP and SV were processed for histochemistry, morphometric-stereological analysis and immunocytochemistry to determine the PCNA index (PI). After 10 days of TR, the VP weight reached approximately 72% of the CT values, while the SV weight exceeded approximately 17% of the CT values. By the third day of TR, VP and SV presented a mean PI of 34% and 94% for distal region and 14% and 22% for proximal region, respectively. SV also had more luminal cells PCNA-positive than VP, mainly in the distal region. The PI values fell on days 5, 7 and 10, but were still higher than CT. These findings indicate that epithelial cells from involuted SV are more responsive to TR than those from VP when stimulated to proliferate and replace the luminal cell population, suggesting a different mechanism regulating cell proliferation in response to androgenic stimuli.

Protein detection in spermatids and spermatozoa of the butterfly Euptoieta hegesia (Lepidoptera)

This study was undertaken to detect protein components in both sperm types of the butterfly Euptoieta hegesia. These spermatozoa possess complex extracellular structures for which the composition and functional significance are still unclear. In the apyrene sperm head, the proteic cap presented an external ring and an internal dense content; basic proteins were detected only in external portions. In the tail, the paracrystalline core of mitochondrial derivatives and the axoneme are rich in proteins. The extratesticular spermatozoa are covered by a proteic coat, which presented two distinct layers. In eupyrene spermatozoa, acrosome and nucleus were negatively stained, probably because of their high compaction. In the tail, there is no paracrystalline core and the axoneme presented a very specific reaction for basic proteins. The lacinate and reticular appendages are composed of cylindrical sub-units and presented a light reaction to E-PTA and a strong reaction to tannic acid. A complex proteic coat also covers the extratesticular spermatozoa. We found similarities between both extratesticular coats, indicating a possible common origin. Both spermatozoon types are rich in proteins, especially the eupyrene appendages and the extratesticular coats. We believe that both coats are related to the sperm maturation and capacitation processes. (AU)

Protein detection in spermatids and spermatozoa of the butterfly Euptoieta hegesia (Lepidoptera)

This study was undertaken to detect protein components in both sperm types of the butterfly Euptoieta hegesia. These spermatozoa possess complex extracellular structures for which the composition and functional significance are still unclear. In the apyrene sperm head, the proteic cap presented an external ring and an internal dense content; basic proteins were detected only in external portions. In the tail, the paracrystalline core of mitochondrial derivatives and the axoneme are rich in proteins. The extratesticular spermatozoa are covered by a proteic coat, which presented two distinct layers. In eupyrene spermatozoa, acrosome and nucleus were negatively stained, probably because of their high compaction. In the tail, there is no paracrystalline core and the axoneme presented a very specific reaction for basic proteins. The lacinate and reticular appendages are composed of cylindrical sub-units and presented a light reaction to E-PTA and a strong reaction to tannic acid. A complex proteic coat also covers the extratesticular spermatozoa. We found similarities between both extratesticular coats, indicating a possible common origin. Both spermatozoon types are rich in proteins, especially the eupyrene appendages and the extratesticular coats. We believe that both coats are related to the sperm maturation and capacitation processes.

Structure and ultrastructure of spermatozoa in Meliponini (stingless bees) (Hymenoptera: Apidae).

In spite of their importance in the reproduction of phanerograms, few bees have received an adequate description of their sperm ultrastructure. In this study, we concluded that it was possible to define a characteristic pattern for Meliponini spermatozoa. However, we have also found species-specific differences that could be used, for example, in a taxonomic or phylogenetic evaluation. Meliponini spermatozoa are made up of a head and a flagellar region. The head includes an acrosome containing the perforatorium, covered by the acrosomal vesicle and a nucleus. In transverse sections, the acrosome is circular at the tip but becomes triangular as it nears the nucleus. The perforatorium base penetrates into a small cavity in the nuclear tip. The flagellum consists of an axoneme, a pair of mitochondrial derivatives, a centriolar adjunct and a pair of accessory bodies. The axoneme has a typical 9+9+2 microtubule pattern. In the final portion, the axoneme becomes gradually disorganized with the accessory microtubules terminating last. The mitochondrial derivatives are asymmetric in length and diameter. In cross-section, they are ellipsoidal and the larger one has a paracrystalline region. The centriolar adjunct begins at the nuclear base and extends parallel to the axoneme up to the anterior end of the smaller mitochondrial derivative.

Protein detection in spermatids and spermatozoa of the butterfly Euptoieta hegesia (Lepidoptera).

This study was undertaken to detect protein components in both sperm types of the butterfly Euptoieta hegesia. These spermatozoa possess complex extracellular structures for which the composition and functional significance are still unclear. In the apyrene sperm head, the proteic cap presented an external ring and an internal dense content; basic proteins were detected only in external portions. In the tail, the paracrystalline core of mitochondrial derivatives and the axoneme are rich in proteins. The extratesticular spermatozoa are covered by a proteic coat, which presented two distinct layers. In eupyrene spermatozoa, acrosome and nucleus were negatively stained, probably because of their high compaction. In the tail, there is no paracrystalline core and the axoneme presented a very specific reaction for basic proteins. The lacinate and reticular appendages are composed of cylindrical sub-units and presented a light reaction to E-PTA and a strong reaction to tannic acid. A complex proteic coat also covers the extratesticular spermatozoa. We found similarities between both extratesticular coats, indicating a possible common origin. Both spermatozoon types are rich in proteins, especially the eupyrene appendages and the extratesticular coats. We believe that both coats are related to the sperm maturation and capacitation processes.

Ultrastructure of apyrene and eupyrene spermatozoa from the seminal vesicle of Euptoieta hegesia (Lepidoptera: Nymphalidae).

The ultrastructure of the seminal vesicle's spermatozoa of the butterfly Euptoieta hegesia was analyzed. The apyrene spermatozoa measure about 300 microm in length and swim freely in a secretion. The anterior end consists in a cap with a cylindrical extension and a globular structure. The flagellum has a 9+9+2 axoneme, two mitochondrial derivatives with paracrystalline matrices and an external coat formed by concentric layers. The eupyrene spermatozoa measure about 550 microm in length and are grouped into bundles. The anterior end consists in an amorphous globule. Posterior to this globule, a coat with a dense material covers the spermatozoon where an acrosome and a nucleus appear. The flagellum has a 9+9+2 axoneme and two mitochondrial derivatives. External to the coat and attached to the dense material, there is a reticular appendage, which has a paracrystalline core and extends to the distal tip of the spermatozoon.

Androgen-dependent synthesis/secretion of caltrin, calcium transport inhibitor protein of mammalian seminal vesicle.

Effects of androgen status on the synthesis and secretion of rat caltrin have been studied by three different procedures: a) immunocytochemistry in seminal vesicle tissues; b) polyacrylamide gel electrophoresis and Western immunostaining of seminal vesicle secretion; and c) evaluation of trypsin inhibitory activity of the seminal vesicle secretion. Rat caltrin has been immunolocalized in cells of the secretory epithelium, specifically in the electron-lucent halo of secretory granules which store and transport proteins to the lumen. No caltrin immunoreaction was detected 14 days postcastration, and the ultrastructure of the epithelial cells was markedly altered. SDS-PAGE and Western blotting of the seminal vesicle secretion revealed alterations in the protein pattern and loss of the caltrin-related immunoreactive bands. The 54-kDa caltrin-precursor protein and the 6.2-kDa active caltrin were absent. Trypsin inhibitory activity of the seminal secretion was reduced about 50% in castrated animals. Daily testosterone administration restored both the protein pattern and immunoreactivity of the seminal vesicle secretion, and, as expected, reversed the morphological alterations of the gland after 7 days of treatment. Trypsin--inhibitor effect of the secretion also returned to normal levels after fourteen days of testosterone administration. Data suggest that the synthesis and secretion of caltrin are testosterone-dependent processes.

Polyamine levels in testes and seminal vesicles from adult golden hamsters during gonadal regression-recrudescence.

The exposure of golden hamsters to short days results in early regression of the reproductive organs and subsequent spontaneous recrudescence characterized by active cellular regeneration and differentiation. Thus, adult male hamsters were subjected to short photoperiod (SP, 6L:18D) for 9, 12, 14, 16, 18, and 22 weeks or maintained under long photoperiod (LP, 14L:10D) for 22 weeks, to assess photoperiodic-related changes in testicular and seminal vesicle (SV) levels of polyamines (PA) that are involved in cell growth and differentiation. During the regression phase, the weights of the organs and the circulating levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH), prolactin, testosterone, dihydrotestosterone, and 5 alpha-androstane-3 alpha, 17 beta-diol were significantly diminished and, thereafter, during the recrudescence phase, they recovered total or partially their control values. In both tissues, the exposure to SP for 14-16 weeks resulted in an increase of PA concentrations, followed by a return to control levels in the recrudescence period. At the time of maximal tissue involution, the ornithine decarboxylase (ODC) activity (key regulatory enzyme of PA biosynthesis) showed a significant increase in testis, preceding the sharp peak of PA concentration. However, a marked decrease in ODC activity was detected in SV. The concentration of N-acetyl PA in SV showed an increment at 16 weeks of SP, while no modifications were detected in testicular concentration. When PA, N-acetyl PA, and ODC activity were expressed per testis and per SV, values fell significantly during the involution period, but in the recrudescence phase levels were recovered concomitantly with the restoration of the organ weight and function. In conclusion, the photoperiodic-related changes in PA and their N-acetyl derivatives might play a crucial role in regrowth and differentiation of the male sexual organs during the spontaneous recrudescence phase. Additionally, organ-specific regulation of the PA biosynthesis pathway could also take place.

Aumento da espessura do epitélio da vesícula seminal do rato, em resposta ao excesso de testosterona plasmática / Increase in the rat seminal vesicle epithelial height in response to excess plasma testosterone level

Investigou-se a resposta morfológica da vesícula seminal do rato adulto ao excesso de testosterona. No período compreendido entre 24 h e 14 dias de tratamento com hCG, os níveis plasmáticos de testosterona exibiram aumento progressivo de duas a cinco vezes os valores basais, enquanto a altura das células secretoras da vesícula seminal apresentou aumento constante de cerca de 20%