Histology, ultrastructure, and seasonal variations in the bulbourethral gland of the African straw-colored fruit bat Eidolon helvum.

We studied the morphological characteristics and seasonal changes of the bulbourethral gland of Eidolon helvum in a typical African tropical environment. Forty-eight bulbourethral glands were examined using gross anatomical, histological, histochemical, and ultrastructural techniques during the early rainy, late rainy, and peak dry seasons. The pear-shaped bilateral bulbourethral glands were located extra-abdominally in the inguinal region. Trabeculae from the capsule divided the parenchyma into numerous lobules of tubuloalveolar glandular acini. The mucosa was covered by a simple columnar epithelium consisting up of principal secretory cells, columnar dense cells and basal cells, which were progressively pronounced during the dry season. The principal cells contained eosinophilic granules, which were PAS positive while the dense cells did not show affinity for the stains. The mean gross weights, acini diameters, and epithelial heights were greater during the rainy season than the dry season. Ultrastructural evaluation showed that the cytoplasm of the principal cells contained well-developed Golgi complexes, rough endoplasmic reticulum, mitochondria, and secretory vesicles of varying electron densities and sizes. The secretory vesicles were numerous during the early rainy season, decreased during the late rainy season and were scanty during the peak dry season. The simple columnar epithelium observed during the rainy season was replaced by an undefined stratified epithelium during the dry season, and this was associated with cellular degenerations and regenerations. In conclusion, E. helvum has a typical mammalian bulbourethral gland, with a unique cell type, the dense cell whose functions are not well-understood. The gland exhibits cyclical seasonal variation in structure and secretory activity; being active during the early rainy season (breeding season), and showing the lowest activity during the dry season (non-breeding season). Glandular epithelial cell renewal occurs during the dry season in preparation for the next breeding season.

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.

Illegal treatment of barrows with nandrolone ester: effect on growth, histology and residue levels in urine and hair.

The effect of 17ß-19-nortestosterone (17ßNT) treatment of barrows on residue levels and growth was evaluated. Five barrows were treated three times during the fattening period with 17ßNT phenylpropionate (Nandrosol, nandrolone phenylpropionate 50 mg/ml,1 mg/kg body weight). Another five barrows were untreated and five boars (untreated) were kept as positive control. Boars and treated barrows showed a 13 and 9% improvement in growth compared to untreated barrows, with mean final body weights of 121.6, 117.8 and 109.0 kg, respectively. The bulbourethral glands of the treated barrows were three times heavier than untreated barrows. The histology of the prostate and bulbourethral gland of the treated barrows was comparable to the boars, whereas the control barrows showed atrophic glands. Levels of 17ßNT ester in hair from treated barrows were high, whereas boars and untreated barrows did not show levels above LLQ. It is concluded that analysis of hair can detect illegal treatment with 17ßNT ester in barrows. The size of the bulbourethral gland can also be used for screening in the slaughterhouse.

Regional distribution of neuroendocrine cells in the urogenital duct system of the male rat.

BACKGROUND: Neuroendocrine (NE) cells are frequently present in the human prostate and urethra, whereas they are lacking in the other urogenital organs. This study was undertaken as there are only few detailed studies available on the distribution, form and function of NE cells and the structure of excretory ducts of the accessory sex organs in the male rat. METHODS: Systematic gross anatomical dissections were combined with immunohistochemical and electron microscopic studies of the excretory ducts of the urogenital glands in male rats, with particular focus on the distribution and ultrastructure of the NE cells. RESULTS: The topography and structure of the excretory ducts of the different glands were characterized in detail and analyzed for the distribution of NE cells. These are present (in falling frequencies) in the ducts of seminal vesicles and ventral and lateral prostate and are rare in ducts of coagulating gland, dorsal prostate, urethral epithelium, and excretory ducts of the (bulbo) urethral glands. They are absent in the respective glands proper, the deferent duct and ejaculatory ampulla. Approximately 40% of the NE cells of the ventral prostate ducts are of the "open" type, whereas these are less frequent (14%) in the seminal vesicle ducts, where the "closed" type prevails. CONCLUSIONS: NE cells are present in unequal quantities in the excretory ducts of the accessory sex glands, but they are absent in the glands proper and the deferent ducts. This distribution pattern points to a strictly localized function and differentiation potency of NE precursor cells.

Structural and ultrastructural features of boar bulbourethral glands.

The morphological features of boar bulbourethral glands were examined by light and transmission microscopy. Bulbourethral glands are compound tubuloalveolar glands surrounded by a capsule of dense connective tissue and arranged in multiple lobules formed by endpieces and excretory ducts. Endpieces and excretory ducts are both lined by a single epithelium of mucous cells with a basal nucleus. Epithelial cells accumulate secretory granules containing neutral and carboxylated acid mucosubstances and a small amount of sulphated acid mucosubstances. The ultrastructure of epithelial cells varies according to the secretory cycle. In initial stages, the cells show a columnar shape and secretory granules unevenly distributed in the cytoplasm. As the synthesis of mucosubstances progresses, the amount of the secretory granules increases and the cellular shape becomes pyramidal. Secretory granules can contain inclusions and present differences among them according to their different phases of formation. In pyramidal cells, secretory products are released into the lumen by a merocrine mechanism.

[Dynamics of proliferation and apoptosis in the human bulbourethral glands in the process of aging].

The dynamics of the processes of proliferation and apoptosis in the epithelium of the bulbourethral glands in men aged 17-90 years was studied using monoclonal antibodies against PCNA and p53. Higher levels of cell reproduction and cell death were noted in the glandular duct epithelium as compared to those in secretory portions. Age-related increase in apoptosis which correlated with the activation of the glandular cell proliferation was shown. In old men, apoptosis in the bulbourethral glands was accompanied by the tendency to reduced cell proliferation. The correlation between the processes of cell reproduction and cell death in the epithelium of the bulbourethral glands, as well as their dependence on blood androgen levels, are discussed.

[Endocrinocytes of the human bulbourethral glands].

Structure, topography and numbers of endocrinocytes in bulbourethral glands of mature men were studied using immunohistochemical demonstration of chromogranin A. Chromogranin-positive cells were found to be predominantly localized in the epithelium of excretory ducts, while they were sparse in the terminal secretory portions. Endocrinocytes in bulbourethral glands were shown to possess argyrophobic properties and to be stained with antibodies against common cytokeratin. The possibility of epithelial histogenesis of bulbourethral gland endocrinocytes is discussed.

Lectin histochemistry of the boar bulbourethral glands.

The present study describes, for the first time, the glycosidic content of boar bulbourethral glands using lectin histochemistry. Fourteen horseradish peroxidase- or digoxigenin-labelled lectins with different carbohydrate specificities were used in samples obtained from 3 healthy Landrace boars. The results obtained indicate that endpiece and duct cells synthesize and secrete mainly O-glycoproteins with alpha- and beta-D-N-acetylgalactosamine, beta-D-galactose-beta(1-->3)-D-N-acetylgalactosamine, D-N-acetylglucosamine and neuraminic acid residues. Glycoproteins secreted by bulbourethral glands have a role in the protection and lubrication of the urethra. In addition, they may be also involved in the regulation of the sperm metabolic activity and in the maintenance of the structural integrity of acrosomal and plasma membranes.

[Fertility alpha-2-microglobulin expression in the human bulbourethral glands]. / Ekspressiia al'fa 2-mikroglobulina fertil'nosti v bul'bouretral'nykh zhelezakh cheloveka.

Fertility alpha-2-microglobulin (FAMG) was demonstrated in the human bulbourethral glands by the method of indirect immunoperoxidase staining using monoclonal antibodies against FAMG. The product of the immunohistochemical reaction was primary found in the duct cells. Weak immunopositive reaction was observed in single epitheliocytes of the secretory units. FAMG-positive granularity was typically found in the supranuclear cytoplasmic zone, that corresponds to the localization of secretory granules both in the glandulocytes and in the duct cells. These results suggest, that the bulbourethral glands are probably the additional producers of human seminal plasma FAMG.

Estudio morfológico de la glándula bulbouretral de conejo oryctolagus cuniculus / Morphologic study of the bulbourethral gland of the oryctolagus cuniculus rabbit

El objetivo del estudio fue conocer la morfología de la glándula bulbouretral de conejo (Oryctolagus cuniculus), para, posteriormente, efectuar el estudio morfofuncional. Se utilizaron 5 conejos (oryctolagus cuniculus), obtenidos del bioterio de la Facultad de Medicina de la Universidad de La Frontera, Chile, durante el período de máxima reproducción. Con la ayuda de una lupa ransor se disecó la región pélvica, retirándose en bloque los componentes anatómicos del aparato genital. Fueron disecadas las estructuras anatómicas cuyos conductos terminaban el la uretra y se aisló la glándula bulbouretral. Las muestras fueron fijadas en Bouin y procesadas para su inclusión en paraplast. Se realizaron cortes seriados de 5 µm de espesor, los cuales fueron teñidas para las observaciones histológicas e histoquímica. El estudio morfológico y las fotografías se realizaron utilizando un microscopio Carl Zeiss, Axiolab, con cámara MC 80 DX. Los resultados mostraron que la glándula bulbouretral del conejo es única, que está rodeada por el músculo bulboglandular y una cápsula fibrosa. De forma cuboidea, alargada en sentido anteroposterior, con ductos cortos que se abren en la pared posterior de la uretra. La glándula se encuentra relacionada con la próstata y parapróstata y está muy vascularizada. Es de tipo alveolar, cubierta de tejidos muscular estriado y conectivo, formado principalmente por fibras colágenas y elásticas, las cuales penetran en su interior dividiéndola en dos lóbulos y subdividiendo cada uno de ellos en lobulillos menores. El estudio histoquímico reveló la presencia de gránulos de glucógeno, mucosustancias neutras y ácidas, sulfatadas y no sulfatadas. La diferencia morfológica observada en la población celular, de la glándula bulbouretral del conejo, se debe, posiblemente, al estado secretorio en que se encuentra

Immunohistolocalization of the carbonic anhydrase isoenzymes (CA-I, CA-II, and CA-III) in the reproductive tract of male horses.

OBJECTIVE: To elucidate locations of cytosolic carbonic anhydrase isoenzyme (CA-I, CA-II, and CA-III)-positive epithelial cells in equine male reproductive organs. DESIGN: Descriptive and immunohistochemical study. ANIMALS: 4 clinically normal male horses. PROCEDURE: The testis (seminiferous tubules, rete tubules), epididymis (initial, middle, and terminal segments), proximal and distal portions of the ductus deferens, ampulla ductus deferentis, seminal vesicle, prostate, and bulbourethral gland were excised from euthanatized horses after administration of an overdose of pentobarbital. The tissue specimens were quickly placed in fixative solution, dehydrated in ethanol, and embedded; then thin sections were cut. For immunohistochemical staining, antibodies against purified equine CA-I, CA-II, and CA-III were raised in rabbits. After examination of the specificity of each antiserum, the monospecific antisera against carbonic anhydrase isoenzymes were used to localize the isoenzymes. RESULTS: Specific staining for CA-III was found in the Sertoli and basal cells of the ductus deferens. Most of the testicular and epididymal tissue, as well as ductus deferens, were virtually negative for the enzymes when stained with the antibody to CA-I and CA-II. In the initial segment of the epididymis, a few principal cells had intense cytoplasmic staining with anti-CA-II. In the male accessory glands, CA-I, CA-II, and CA-III were detected in the epithelial cells of the seminal vesicle, prostate, and bulbourethral gland. CONCLUSIONS: In the equine male reproductive tract, the bicarbonate in semen originates mainly from accessory reproductive glands. All 3 isoenzymes may have central roles in the regulation of bicarbonate concentration in seminal plasm and, accordingly, regulate seminal plasma pH. Distribution of CA-III in Sertoli and basal cells of the ductus deferens suggests other specialized physiologic roles.

Effect of mesenchymal glandular inductors on the growth and cytodifferentiation of neonatal mouse seminal vesicle epithelium.

To investigate the developmental properties of glandular mesenchymal inductors along the cranial-caudal extent of the developing male urogenital tract, neonatal mouse seminal vesicle epithelium (SVE) was combined with mesenchyme of the seminal vesicle (SVM), urogenital sinus (UGM), bulbourethral gland (BUG-M), or bladder (BLM) and grafted under the renal capsule of adult syngeneic or athymic male mice. Both SVM + SVE and UGM + SVE tissue recombinants expressed SV histogenesis and SV secretory proteins. BUG-M + SVE recombinants exhibited extensive growth as evidenced by a 36-fold increase in wet weight and a 27-fold increase in DNA content; however, the glandular structures that were induced in the SVE lacked the convoluted mucosa typical of SV. Furthermore, neither SV nor prostatic secretory proteins were detected in these recombinants. SVE grown in association with BLM failed to develop altogether. Thus, the ability to promote SV histogenesis and function is distinctly different in mesenchyme of cranial (SVM and UGM) versus caudal (BUG-M) regions. This implies the existence of a glandular inductive field in the developing male urogenital tract within which inductive activity varies regionally.

Diffuse lymphoid tissue associated with the human bulbourethral gland. An immunohistologic characterization.

This study investigates the presence and distribution of immunocompetent cells in bulbourethral glands obtained from four multiorgan transplant donors. Monoclonal antibodies reacting with B (Leu 12+) and T (Leu 4+) cells, suppressor-cytotoxic cells (Leu 2+), helper-inducer (Leu 3a+) and natural killer (Leu7+) phenotypes, monocyte-macrophages, (LeuM3+), and cells expressing interleukin-2 receptor and HLA-DR antigen were tested in all specimens using an indirect immunoperoxidase staining procedure. T lymphocytes were estimated to represent 10% of the mucosal cell population. Almost all intraepithelial lymphocytes were suppressor-cytotoxic (CD8+) cells. The results demonstrate the presence of a defined distribution of immunocompetent cells in these sex accessory glands. Their role in combatting infections or other chronic genitourinary diseases is still undefined.

Morfologia e histoquímica de glicogênio e mucossubstâncias na próstata e glândulas bulbo-uretrais de Marmosa cinerea Temminck, 1824 (Marsupialia - Didelphidae) / Morphology, glycogen and mucosubstances histochemistry of the prostate and bulbo-urethral glands of the Marmosa cinerea Temminck, 1824 (Marsupialia - Didelphidae)

Sao descritos os aspectos histológicos e a distribuiçao de glicogênio e mucossubstâncias na próstata e nos três pares de glândulas bulbo-uretrais (Cowper) de Marmosa cinerea, marsupial Didelphidae com ampla distribuiçao geográfica no Brasil. Os três segmentos prostáticos, cranial, médio e caudal, apresentam túbulos secretores disseminados na mucosa uretral e sao divididos em zonas externa, média e interna, de acordo com as características morfológicas e tintoriais do epitélio secretor. No segmento cranial, o epitélio secreta glicogênio e mucossubstâncias neutras. No segmento médio sao produzidas mucossubstâncias neutras e sialomucinas, sendo sugerido um mecanismo apócrino de secreçao neste segmento. O epitélio do segmento caudal produz glicogênio, mucossubstâncias neutras e proteínas. O par lateral de bulbo-uretrais é formado por túbulos secretores longos e ramificados, que produzem mucossubstâncias neutras. O par intermédio constitui-se de túbulo-ácinos revestidos por células com porçao apical expandida e que secretam mucossubstâncias neutras, sialomucinas e sulfomucinas. Os túbulo-ácinos do par medial sao tortuosos e suas células produzem mucossubstâncias neutras.

Basal cells of H-Dunning tumor are myoepithelial cells. A comparative immunohistochemical and ultrastructural study with male accessory sex glands and mammary gland.

Recent immunohistochemical studies have shown that basal cells in human prostatic epithelium are not myoepithelial cells. Since in the literature the Dunning tumor, originally described as a rat prostate carcinoma derived from the dorsolateral prostate of a Copenhagen rat, was reported to have myoepithelial cells, a comparative immunohistochemical and ultrastructural study was performed in the H-, HIF- and AT3-lines of the Dunning tumor, the male accessory sex glands (ventral, dorsal, lateral prostate, coagulating gland, bulbourethral gland) and the mammary gland of both Copenhagen and Wistar rats. Mono- and polyclonal antibodies directed against intermediate filament proteins (cytokeratin, desmin, vimentin) and the contractile proteins (alpha-actin, muscle type specific myosin, tropomyosin) were used along with phalloidin decoration of F-actin. As in the human prostate, none of the rat prostate lobes in either strains did contain basal cells expressing cytokeratin along with alpha-actin, myosin and tropomyosin Cells representing fully differentiated myoepithelial cells, however, were present as anticipated in the mammary gland, the bulbourethral gland and the H-tumor line of the Dunning tumor. This finding is difficult to reconcile with the contention of a prostatic origin of the H-Dunning tumor. Further studies are required to classify the epithelial parental tissue in order to define the true origin of the H-Dunning tumor and the tumor lines derived thereof.

Inhibitory effects of technical grade methoxychlor on development of neonatal male mouse reproductive organs.

Effects of technical grade methoxychlor (MX), an estrogenic insecticide, and 17 beta-estradiol (E2) were examined on serum testosterone (T) concentrations and growth and histology of neonatal male mouse reproductive organs. Male NIH/Swiss mice received i.p. injections daily from birth to Day 9 with one of the following: 10 micrograms E2 or 0.1 or 1.0 mg MX or sesame oil vehicle. The mice were killed on Day 10. MX or did not affect body weights or mortality. Serum T concentrations in control mice were 0.29 +/- 0.03 ng/ml; T concentrations in males treated with MX or E2 were reduced. The 1.0-mg dose of both MX and E2 significantly decreased DNA contents of the seminal vesicles (SV), bulbourethral glands (BUG), and ventral prostate (VP) compared to controls. In the same animals, DNA contents of testes, epididymides, and efferent ductules were not decreased. The lower dose of MX (0.1 mg) decreased DNA content of only BUG and SV. DNA content of the ductus deferens was not affected by any treatment. E2 and both doses of MX also decreased epithelial morphogenesis in the SV and BUG, and inhibited the onset of mucin production in BUG epithelium and smooth muscle differentiation in the ductus deferens. In summary, our results indicate that technical grade MX, at doses as low as 0.1 mg/day, and E2 inhibit neonatal male reproductive tract development and decrease serum T concentrations.

Anti-androgenic effects of Win 49,596 on development of the neonatal mouse bulbourethral gland in culture.

The effects of the anti-androgen Win 49,596, a steroidal androgen receptor antagonist, on the testosterone (T) dependent development of neonatal mouse bulbourethral glands (BUGs) were examined in vitro. Day of birth (day 0) BUGs were grown in organ culture for 6 days in Dulbecco's modified Eagle's medium:Ham's F-12 media (1:1, volume/volume) with 10% fetal calf serum. Cultures were grown with or without T (10 nM) and various concentrations of Win 49,596 (0-57.6 microns). The DNA content of BUGs grown with T alone increased 4-fold over the culture period and the epithelium grew and branched extensively; without T only minimal growth and epithelial morphogenesis occurred. In the presence of T, Win 49,596 inhibited development in a dose-dependent manner, with an ED50 of 0.8 microM; concentrations at or above 3.6 microM produced maximal inhibition of development. Win 49,596 alone at 14.4 microM did not stimulate BUG growth, demonstrating a lack of agonist activity. BUGs grown for 3 days with Win 49,596 and T, then an additional 6 days with only T, did not resume development. In summary, Win 49,596 produced a dose-dependent suppression of BUG development in vitro, and was not androgenic. Additionally, the inhibitory effects of Win 49,596 persist for at least 6 days following cessation of treatment.

Reproductive biology of the male bent-winged bat, Miniopterus schreibersii (Vespertilionidae) in southeast South Australia.

The seasonal chronology of the events of the reproductive cycle, and changes in the structure and function of the primary and accessory organs of the male bent-winged bat, Miniopterus schreibersii, were studied at latitude 37 degrees S in temperate southeastern Australia. The testicular cycle commenced in late spring (November), and sperm appeared in the seminiferous tubules and epididymides in early fall (March). The cycle of the accessory sex gland complex generally paralleled the testicular cycle, reaching maximum hypertrophy at the time of insemination in late fall (April/May). Thereafter, the primary and secondary sex glands (except the ampullary gland) involuted as the animals entered winter torpor. However, a cauda epididymal store of sperm persisted until late spring, and sperm were often observed, as well, in the ampullary gland duct and alveoli throughout winter. This study has confirmed that male Miniopterus differs from other vespertilionids in that accessory gland activity declines following the fall breeding in keeping with the fact that, unlike in other vespertilionids, insemination, ovulation and conception are concurrent events in the fall in this species. The reduced secretory status of the Leydig cells and exceptionally low levels of circulating androgens throughout the year, in combination with the presence of viable epididymidal sperm for most of gestation, are all interesting features of this reproductive cycle.

Immunoglobulins and immunoglobulin-containing cells in the reproductive tract of normal rams.

The levels of the immunoglobulins IgA, IgG1, IgG2, and IgM were measured in serum and fluid from various locations in the reproductive tract of normal rams. These fluids included semen, preputial washings, and fluid from the accessory sex glands (ASG), vasa deferens, rete testes, and tissue fluid from the seminal vesicles, bulbourethral glands, epididymal tails and efferent ducts. In addition, the prevalence of specific Ig-containing cells (ICC) was measured in sections of formalin fixed tissues stained by an indirect peroxidase-antiperoxidase labelling technique. Mean IgA levels in semen (1.23 mg/ml) and ASG fluid (0.46 mg/ml), were higher than in serum (0.19 mg/ml) and were at levels higher than IgG1 or IgG2 levels in semen, ASG fluid, and preputial washings, thus confirming the existence of a local immune system primarily in the ASG of ram genitalia. Relatively low concentrations of IgA and IgG in other genital fluids and IgG levels in these fluids were consistent with diffusion from serum. The relatively high prevalence of IgA-containing cells in bulbourethral (56% of all ICC) and prostate (49%) glands confirmed these tissues as major sites of local Ig production. ICC were also found in large numbers beneath pelvic urethral and preputial epithelia, but these were predominantly IgG-containing (88 and 72% respectively).