Seminal vesicle invasion on multi-parametric magnetic resonance imaging: Correlation with histopathology.

OBJECTIVES: The pre-treatment risk of seminal vesicle (SV) invasion (SVI) from prostate cancer is currently based on nomograms which include clinical stage (cT), Gleason score (GS) and prostate-specific antigen (PSA). The aim of our study was to evaluate the staging accuracy of 3T (3T) multi-parametric (mp) Magnetic Resonance Imaging (MRI) by comparing the imaging report of SVI with the tissue histopathology. The additional value in the existing prediction models and the role of radiologists' experience were also examined. METHODS: After obtaining institutional review board approval, we retrospectively reviewed clinico-pathological data from 527 patients who underwent a robot-assisted radical prostatectomy (RARP) between January 2012 and March 2015. Preoperative prostate imaging with an endorectal 3T-mp-MRI was performed in all patients. Sequences consisted of an axial pre-contrast T1 sequence, three orthogonally-oriented T2 sequences, axial diffusion weighted and dynamic contrast-enhanced sequences. We considered SVI in case of low-signal intensity in the SV on T2-weighted sequences or apparent mass while diffusion-weighted and DCE sequences were used to confirm findings on T2. Whole-mount section pathology was performed in all patients. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of MRI (index test) for the prediction of histological SVI (reference standard) were calculated. We developed logistic multivariable regression models including: clinical variables (PSA, cT, percentage of involved cores/total cores, primary GS 4-5) and Partin table estimates. MRI results (negative/positive exam) were then added in the models and the multivariate modeling was reassessed. In order to assess the extent of SVI and the reason for mismatch with pathology an MRI-review from an expert genitourinary radiologist was performed in a subgroup of 379 patients. RESULTS: A total of 54 patients (10%) were found to have SVI on RARP-histopathology. In the overall cohort sensitivity, specificity, PPV and NPV for SVI detection on MRI were 75.9%, 94.7%, 62% and 97% respectively. Based on our sub-analysis, the radiologist's expertise improved the accuracy demonstrating a sensitivity, specificity, PPV and NPV of 85.4%, 95.6%, 70.0% and 98.2%, respectively. In the multivariate analysis PSA (odds ratio [OR] 1.07, p=0.008), primary GS 4 or 5 (OR 3.671, p=0.007) and Partin estimates (OR 1.07, p=0.023) were significant predictors of SVI. When MRI results were added to the analysis, a highly significant prediction of SVI was observed (OR 45.9, p<0.0001). Comparing Partin, MRI and Partin with MRI predictive models, the areas under the curve were 0.837, 0.884 and 0.929, respectively. CONCLUSIONS: MRI had high diagnostic accuracy for SVI on histopathology. It provided added diagnostic value to clinical/Partin based SVI-prediction models alone. A key factor is radiologist's experience, though no inter-observer variability could be examined due to the availability of a single expert radiologist.

Analysis of the cytoskeleton organization and its possible functions in male earthworm germ-line cysts equipped with a cytophore.

We studied the organization of F-actin and the microtubular cytoskeleton in male germ-line cysts in the seminal vesicles of the earthworm Dendrobaena veneta using light, fluorescent and electron microscopy along with both chemically fixed tissue and life cell imaging. Additionally, in order to follow the functioning of the cytoskeleton, we incubated the cysts in colchicine, nocodazole, cytochalasin D and latrunculin A. The male germ-line cells of D. veneta are interconnected via stable intercellular bridges (IB), and form syncytial cysts. Each germ cell has only one IB that connects it to the anuclear central cytoplasmic mass, the cytophore. During the studies, we analyzed the cytoskeleton in spermatogonial, spermatocytic and spermatid cysts. F-actin was detected in the cortical cytoplasm and forms distinct rings in the IBs. The arrangement of the microtubules changed dynamically during spermatogenesis. The microtubules are distributed evenly in whole spermatogonial and spermatocytic cysts; however, they primarily accumulate within the IBs in spermatogonia. In early spermatids, microtubules pass through the IBs and are present in whole cysts. During spermatid elongation, the microtubules form a manchette while they are absent in the cytophore and in the IBs. Use of cytoskeletal drugs did not alter the general morphology of the cysts. Detectable effects-the occurrence of nuclei in the late spermatids and manchette fragments in the cytophore-were observed only after incubation in nocodazole. Our results suggest that the microtubules are responsible for cytoplasmic/organelle transfer between the germ cells and the cytophore during spermatogenesis and for the positioning of the spermatid nuclei.

Morphology and biometric of the vesicular and bulbourethralglands in castrated and non-castratedSanta Inesbreed sheep

Introduction: The purpose of this paper is to determine the topography, biometry and light microscopyimage of the vesicular and bulbourethral glands in order to analyze morphologic features of the accessorysexual glands in castrated and non-castrated animals.Materials and Methods:The morphology of theaccessory sexual glands was investigated in 14 adult Santa Inesbreed sheep, weighing 32 kg, on average.Six of them were castrated, and eight, non-castrated. For macroscopic study, the description of these twoglands was carried out, as well as dissection and biometry study. Moreover, weight, length, height andwidth measurements were evaluated. For histological analysis, the vesicular and bulbourethral glands weresampled.Results:The topography of the reproductive glands was similar to bovine species. However, lowermacroscopic measurements (p < 0,05) in the glands of the castrated sheep were evidenced when comparedwith the non-castrated ones. Characteristics such as shape of the glands, composition of the layer mucosa,the lamina propria, muscular, the excretory ducts and the adventitia were determined.Conclusion:Sheepcastration promoted changes in the biometric measures of the glands, which were lower in castrated animals.The morphological and biometric characteristics of the vesicular and bulbourethral glands in sheep weredetermined.

Genesis of spermatodesms in Tylopsis liliifolia (Orthoptera: Phaneropterinae) and their transit in the male genital tract.

The spermatodesms of Tylopsis liliifolia form in the most proximal follicular cysts and are composed of a large number of sperm held together by a cap located in the anterior region of the acrosome. The cap is formed by short thin fibrils, loosely arranged at random, probably derived from secretory activity of cells of the cyst wall. Compared to other Tettigoniidae, a peculiar feature is acrosomal wings that twist gradually around the anterior region of the nucleus; at the end of the twisting process, the region of the sperm acrosome, observed in cross section, shows a typical spiral form. Spermatodesms do not undergo any substantial changes in the spermiduct. The epithelial cells of the wall have secretory activity and many show marked spermiophagic activity, which is conducted by epithelial cell protrusions that envelop the gametes, taking them into the cytoplasm. When removed from seminal vesicles and observed in vivo, spermatodesms show accentuated corkscrew movement, and when observed by SEM, slight torsion. Thus organized, spermatodesms are transferred to the spermatophore during mating, where they are transformed before reaching the seminal receptacle.

Immature rat seminal vesicles show histomorphological and ultrastructural alterations following treatment with kisspeptin-10.

BACKGROUND: Degenerative effects of critical regulators of reproduction, the kisspeptin peptides, on cellular aspects of sexually immature male gonads are known but similar information on accessory sex glands remain elusive. METHODS: Prepubertal laboratory rats were injected kisspeptin-10 at three different dosage concentrations (10 pg, 1 ng and 1 microgram) for a period of continuous 12 days at the rate of two doses per day. Control rats were maintained in parallel. The day following the end of the experimental period, seminal vesicles were removed and processed for light and electron microscopic examination using the standard methods. DNA damage was estimated by DNA ladder assay and DNA fragmentation assay. RESULTS: The results demonstrated cellular degeneration. Epithelial cell height of seminal vesicles decreased significantly at all doses (P < 0.05). Marked decrease in epithelial folds was readily noticeable, while the lumen was dilated. Ultrastructural changes were characterized by dilatation of endoplasmic reticulum and Golgi complex, heterochromatization of nuclei, invagination of nuclear membranes and a decreased number of secretory granules. Percent DNA damage to the seminal vesicle was 19.54 +/- 1.98, 38.06 +/- 2.09 and 58.18 +/- 2.59 at 10 pg, 1 ng and 1 microgram doses respectively. CONCLUSION: The study reveals that continuous administration of kisspeptin does not lead to an early maturation but instead severe degeneration of sexually immature seminal vesicles.

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.

In situ hybridization and immunohistochemical localization of leptin hormone and leptin receptor in the seminal vesicle and prostate gland of adult rat.

The role of leptin in the regulation of male reproductive function is still a matter of debate. Knowledge about a possible source of leptin in the seminal plasma may therefore be helpful in identifying and elucidating the physiological role of leptin hormone in male reproduction. In our investigation, the expression of leptin and its long receptor isoform (Ob-Rb) was studied in adult male Wistar rats using RT-PCR, Southern blot, in situ hybridization and immunohistochemistry. RT-PCR analysis revealed the expression of both leptin and its Ob-Rb in the seminal vesicle and prostate gland. In situ hybridization also localized the mRNA transcripts of leptin and Ob-Rb in the glandular secretory epithelial cells of prostate gland and seminal vesicle. Immunohistochemistry detected the leptin hormone in the lining epithelium of both male genital glands. In conclusion, these findings suggest that the seminal vesicle and prostate gland could be the possible sources of leptin in the seminal plasma. This leptin might have a direct (paracrine, autocrine or both) effect on epithelial cells of the accessory male genital glands, on the spermatozoa via spermatozoan leptin receptors.

Estudio morfológico de la próstata y glándulas vesiculares de cobayo (Cavia porcellus) / Morphologic study of the prostate and vesicular glands of the guinea pig (Cavia porcellus)

El objetivo de esta investigación fue conocer la morfología de la próstata y glándula vesicular del cobayo (Cavia porcellus), describiéndose aspectos mesoscópicos, histológicos e histoquímicos. Se utilizaron 5 cobayos machos adultos (Cavia porcellus) sanos y alimentados con pellets, zanahorias y frutas ad libitum, en el Bioterio de la Universidad de La Frontera. Una vez sacrificados los animales, fueron aisladas las estructuras cuyos conductos terminaban en la uretra (conductos deferentes, glándulas vesiculares, glándulas coaguladoras, próstata y glándulas bulbouretrales). La próstata y las glándulas vesiculares fueron fijadas en formalina tamponada durante 24 horas y procesadas para su inclusión en paraplast. Se realizaron cortes seriados de 4 µm de grosor, los cuales fueron teñidos para los estudios histológico e histoquímicos. El análisis se realizó a través de un microscopio Carl Zeiss, Axiolab, con cámara Cannon, G6. La próstata se encuentra dorsal a la uretra y está constituída por una masa de tejido glandular cubierto por una delgada cápsula de tejido fibroso y células musculares lisas. Presenta dos lóbulos, derecho e izquierdo, unidos por un istmo dorsal a la uretra, por la cara ventral de cada lóbulo emergen de 8 a 10 pequeños conductos excretores. Histológicamente, está compuesta por unidades túbulo-alveolares recubiertos por epitelio secretor simple cúbico. Las glándulas vesiculares son dos estructuras túbulo-lobulares. Se ubican dorsal a la uretra, relacionándose ventralmente con los conductos deferentes y la cara dorsal de la vejiga. Histológicamente, presentan una capa mucosa con un epitelio cilíndrico de tipo secretor; una capa media constituida principalmente por tejido muscular liso y una capa externa serosa. La histoquímica, en ambas glándulas fue negativa para glucógeno, mucinas neutras y ácidas, sulfatadas y no sulfatadas, tanto en el tejido glandular como en su producto de secreción. La próstata y glándulas vesiculares del cobayo, guardan...

The objective of this investigation was to study the morphology of the prostate and vesicular gland of the guinea pig (Cavia porcellus), describing the mesoscopic, histological and histochemical aspects. Five healthy adult male guinea pigs (Cavia porcellus) were used. The animals received pellets, carrots and fruit ad libitum in the Biotherium of the Universidad de La Frontera. Once the animals were sacrificed the structures which ducts ended in the urethra, (vas deferens, vesicular glands, coagulator glands, prostate and bulbourethral glands) were isolated. The prostate and vesicular gland were fixed in buffered formalin during 24 hours and processed for their inclusion in paraplast. Serial cuts 4 µm thick were realized and stained for histological and histochemical studies, using a Carl Zeiss, Axiolab microscope with a Cannon G6 camera. The prostate is located dorsal to the urethra and is constituted by a mass of glandular tissue covered by a thin capsule of fibrous tissue and smooth muscular cells presenting two lobes, right and left joined to the urethra by a dorsal isthmus. Emerging through the ventral surface of each lobe are 8 to 10 small excretory ducts. Histologically it is composed by alveolar tubular units covered by a simple cubical secretor epithelium. The vesicular glands are two tubular lobular structures located dorsal to the urethra and are connected ventrally with the vas deferens and the dorsal surface of the bladder. Histologically presenting a mucous layer with a secretor type cylindrical epithelium, a medium layer namely constituted by smooth muscular tissue and a serous external layer. Histochemical reaction in both glands was negative for glycogen, for neutral and acid mucins, both sulfate and non sulfate, in the glandular tissue as well as the secretor product. The prostate and vesicular glands of the guinea pig are related to morphological aspects of other mammals. However, the differences found in the histochemical results suggest t...

Malignancy arising in seminal vesicles in the transgenic adenocarcinoma of mouse prostate (TRAMP) model.

BACKGROUND: Transgenic adenocarcinoma of mouse prostate (TRAMP) mice, derived by prostate specific expression of SV40 large T antigen using the rat probasin promoter, all develop prostate tumors akin to human prostate cancers. More recently, epithelial-stromal (ES) tumors resembling phyllodes tumors have been described in the seminal vesicles of TRAMP mice. We report malignancy arising in these ES tumors of the seminal vesicles in TRAMP mice. METHODS: H&E stained sections from 28-week-old TRAMP mice autopsies were examined. Immunostains (cytokeratin, vimentin, desmin, and MIB-1) and electron microscopy were performed on selected blocks of the genitourinary system and metastatic tumor nodules. RESULTS: The seminal vesicles frequently develop tumors containing broad papillae, with bland epithelium and a cellular spindled stroma just beneath the epithelium. The stromal cells have high nuclear to cytoplasmic ratio, frequent apoptotic cells and mitoses. In some cases, the stromal cells become large mass lesions that overgrow the prostate. The epithelium can also proliferate and become malignant. The tumors have high proliferation indices by MIB-1. Some metastatic tumors have characteristics similar to the seminal vesicle ES tumor. CONCLUSIONS: Metastatic tumors in TRAMP mice show three patterns: (1) A definite adenocarcinoma pattern metastatic from the prostate; (2) poorly differentiated tumor without epithelial differentiation; (3) carcinosarcomatous pattern. The carcinosarcomatous pattern and some of the poorly differentiated tumors likely arise from seminal vesicle ES tumors.

The effects of electromagnetic field on the microstructure of seminal vesicles in rat: a light and transmission electron microscope study.

In the industrial world, almost everyone is unavoidably exposed to ambient electromagnetic field (EMF) generated from various technical and household appliances. Controversy exists about the effects of EMF on various tissues of the living bodies. Seminal vesicles as one of these accessory glands play an important role in natural seminal fluid formation and the effects of EMF on its tissue is worthy of investigation. In order to examine this 30 rat were selected and kept for one weeks in quarantine and 15 (experimental group) were exposed to 50 Hz (non-ionizing radiation) during postnatal life for 2 months. The materials were processed and observed under a light and transmission electron microscope. In the experimental rats epithelial and basal cells showed significant destructions presented by heterochromatin and dense nuclei. Cell debris and abnormal areas was recognizable in the stromal connective tissue. Obvious vacuolization was present within the epithelial cell cytoplasm and also between the cellular organelles. The nuclei of the endothelial cells of the blood vessels were more rigid and endothelial cell cytoplasm contained a lot of vacuoles and pinoctotic vesicles. The results suggested that EMF exposure may cause profound changes in the vesicle seminal tissues. Therefore exposure to EMF may result in pathological changes that lead to sub fertility and infertility.

Morphology of male reproductive system in three species of Trypoxylon (Trypargilum) Richards (Hymenoptera: Crabronidae) / Morfologia do sistema reprodutor masculino em três espécies de Trypoxylon (Trypargilum) Richards (Hymenoptera: Crabronidae)

Variations in the adult male reproductive system among different groups of Hymenoptera offer characteristics that help studies on behavior and phylogenetics. The objective of this study was to describe the adult male reproductive system of three Trypoxylon (Trypargilum) species. For that, tissues were disseced, fixed in 2.5 percent glutaraldehyde in 0.1 M sodium cacodylate buffer, pH 7.2 and postfixed in 1 percent osmium tetroxide. The material was dehydratated and embedded for light and electron transmission microscopes. The species have similar reproductive systems, which are formed by a pair of testes, each one with three fusiforme follicles, from which emerges an efferent duct that later joins forming a deferent duct. The deferent duct opens into an ejaculatory duct. The first half of the deferent duct is enlarged and differentiated in a region specialized in sperm storage, the seminal vesicle. The accessory gland flows in the post-vesicular region of the deferent duct. The testes and vesicles are both covered with a conjunctive capsule. Sexually mature individuals have all spermatogenesis stages in their follicles. Sperms are released from testes in bundles which are disorganized inside seminal vesicles.

Variações no sistema reprodutor entre os diferentes grupos de Hymenoptera oferecem caracteres que auxiliam nos estudos de comportamento e filogenia. O objetivo deste trabalho foi descrever o sistema reprodutor masculino de três espécies de Trypoxylon (Trypargilum). Para isso, os tecidos foram dissecados, fixados em glutaraldeído 2,5 por cento em tampão cacodilato de sódio 0,1 M, pH 7,2 e pós-fixados em tetróxido de ósmio a 1 por cento. O material foi desidratado e incluído para microscopias de luz e eletrônica de transmissão. As espécies possuem os sistemas reprodutores muito semelhantes, formados por um par de testículos, cada um com três folículos fusiformes, a partir dos quais emerge um ducto eferente que depois se juntam formando o ducto deferente. O ducto deferente termina no ducto ejaculatório. A primeira metade dos ductos deferentes é dilatada e diferenciada em uma região especializada no armazenamento de espermatozóides, a vesícula seminal. A glândula acessória desemboca na região pós-vesicular do ducto deferente. Testículos e vesículas seminais são envoltos por uma única cápsula conjuntiva. Indivíduos maduros sexualmente apresentam todos os estágios da espermatogênese em seus folículos. Os espermatozóides são liberados dos testículos em feixes, os quais estão desorganizados na vesícula seminal.

Distribution and role of myofibroblasts in human normal seminal vesicle stroma.

We studied the distribution of myofibroblasts in the stroma of normal seminal vesicles. Twelve normal seminal vesicles obtained by surgery on the diagnosis of some diseases were selected, and we evaluated the distribution of myofibroblasts in the seminal vesicles using immunohistochemical and electron and immunoelectron microscopic techniques. Immunohistochemically, myofibroblasts, which were positive for alpha-smooth muscle actin (ASMA) and negative for high molecular weight caldesmon (h-CD), were observed in the stroma just beneath the epithelium of normal seminal vesicles. Moreover, an electron microscope examination revealed the presence of spindle or stellate cells in the stroma of the lamina propria beneath the seminal vesicle epithelium, and an immunoelectron microscopic examination showed that these cells were positive for ASMA. Finally, myofibroblasts are distributed in the lamina propria of human normal seminal vesicles and may play an important role in the ejection of sperm plasm.

Structural and ultrastructural features of boar seminal vesicles.

The morphological features of boar seminal vesicles were examined by light and transmission microscopy. Boar seminal vesicles consist of glandular tissue arranged in multiple lobules containing a system of ramified secretory tubules. The secretory tubules are composed of a mucosa formed by an epithelium and an underlying lamina propria and, are surrounded by a muscular layer. The epithelium is made up of columnar cells and occasional basal cells. Mast cells are frequently found among epithelial cells. Three types of columnar cells, considered different stages of the secretory cell cycle, are present: principal cells, clear cells and dense cells. Principal cells are functionally differentiated cells characterised by abundant mitochondria, great development of the rough endoplasmic reticulum and presence of secretory granules in their cytoplasm. The apical surface of many principal cells shows apical blebs filled with PAS-positive material. No acid mucosubstances are detected. Microvilli cover the apical surface except in the apical blebs. Dense cells, arranged between principal cells, are also functional differentiated cells but with signs of cellular degeneration. Clear cells are an initial differentiated stage of columnar cells and are characterised by the presence of a poorly developed rough endoplasmic reticulum and by the absence of secretory granules. Proliferating cells are present among columnar cells. Basal cells contain scarce cytoplasm, few organelles and no secretory granules. The lack of mitotic activity in these cells suggests that they do not act as precursors of columnar cells.

Immunocytochemical distribution of nitric oxide synthase in the human seminal vesicle: a light and electron microscopical study.

Although nitric oxide (NO) has been proven to be one of the most important non-adrenergic, non-cholinergic mediators in the control of human reproductive tract organs, to date information on the significance of NO-mediated signal transduction in the control of human seminal vesicle (SV) function is still sparse.()Recent investigations have underlined the significance of NO in the maintenance of sperm capacitation and viscosity of the seminal plasma as well as in the control of mammalian seminal vesicle smooth muscle tone. In order to further investigate the functional impact of NO on the regulation of normal SV function, we examined the distribution of NADPH-diaphorase (NADPH-d), endothelial nitric oxide synthase (eNOS) and neuronal nitric oxide synthase (nNOS) in the cellular anatomy of human SV by means of light and electron microscopical immunocytochemistry (LM, EM) in combination with the tyramide signal amplification technique. Human SV were obtained from 15 patients who had undergone surgery for pelvic malignancies (carcinoma of the prostate or urinary bladder). SV specimens were fixed, sectioned and examined by LM and EM for the presence of NAPDH-d, eNOS and nNOS using specific antibodies and advanced staining procedures. LM revealed a dense NADPH-d reaction in glandular epithelial structures, whereas no substantial labeling was detected in the fibromuscular stroma. EM showed that the NADPH-d reaction product was abundantly detectable attached to membranes of the endoplasmic reticulum, mitochondria and the nuclei of glandular epithelial cells. nNOS staining was found in nerve fibers branching within the SV tissue. eNOS staining was present in small vessels but was only observed to a minor degree in glandular and subglandular structures and the smooth muscle stroma. Our results support the hypothesis that human SV is a site of NO production. The distribution of NADPH-d may give rise to the speculation that NO is mainly involved in the regulation of SV secretory activity. The sparse correlation between NADPH-d-, eNOS- and nNOS-staining might hint at the existence of a previously unidentified NOS isoform in human SV.

Effects of diadenosine polyphosphates and seminal fluid vesicles on rabbit sperm cells.

Membrane vesicles were isolated from rabbit seminal plasma. Electron microscopy analyses showed the presence of numerous small, round vesicles with a diameter of about 70 nm. Determination of enzyme activities was carried out by high performance liquid chromatography and showed that the vesicles can degrade the diadenosine polyphosphates (ApnA), Ap3A and Ap4A and ATP and ADP, but not AMP. Studies of the degradation of diadenosine compounds by the vesicles present in seminal fluid showed an increasing production of AMP as the by-product and a time-dependent generation of dephosphorylated products consistent with the presence of ecto-ATP diphosphophosphatase (ecto-apyrase). In the presence of rabbit spermatozoa, AMP did not accumulate because 5'nucleotidase and adenosine deaminase, present at the surface of sperm cells, transformed AMP into adenosine and inosine. The effects of seminal fluid vesicles and diadenosine compounds on the acquisition of fertilizing capacity by rabbit spermatozoa were evaluated by Pisum sativum agglutinin fluorescein isothiocyanate conjugated staining. The results obtained with uncapacitated spermatozoa showed that the capacitating effector BSA could be substituted efficiently by the addition of diadenosine compounds and vesicles previously incubated for 2 h to the capacitative medium. Under these experimental conditions, the spontaneous acrosome reaction rate was not increased. Capacitated rabbit spermatozoa did not undergo acrosome reaction when l-alpha-lysophosphatidylcholine was substituted by diadenosine compounds previously incubated with vesicles. In conclusion, this study has shown that rabbit seminal fluid vesicles can degrade diadenosine compounds to AMP and that the addition of the vesicles and diadenosine compounds to uncapacitated rabbit spermatozoa favours the acquisition of the fertilizing capacity.

Mesoscopía e Histología de la glándula vesicular en el conejo (Oryctolagus cuniculus) / Mesoscopy and histology of the vesicular gland in the rabbit (Oryctolagus cuniculus)

El objetivo de este estudio fue conocer con mayor detalle la morfología de la glándula vesicular del conejo Oryctolagus ciniculus para, posteriormente efectuar estudios morfofuncionales. Se utilizaron 12 conejos machos (Oryctolagus cuniculus) clínicamente sanos, mantenidos en el Bioterio de la Facultad de Medicina de la Universidad de La Frontera, Temuco, Chile. Sacrificados los conejos, se registró su peso corporal y se disecó la región pélvica, retirando en bloque los componentes anatómicos del aparato urogenital, aislando posteriormente la glándula vesicular de las otras glándulas anexas. Las muestras fueron fijadas en formalina tamponada y procesadas para su inclusión en paraplast. Se realizaron cortes seriados de 4 µ de espesor, los cuales fueron teñidos para su estudio histológico e histoquímico. Para el análisis morfológico y las fotografías se utilizó un microscopio Carl Zeiss, Axiolab con cámara MC 80 DX. Los resultados mostraron que la glándula vesicular del conejo macho (Oryctolagus cuniculus) presentó una forma irregular, globosa cranealmente y aplastada caudalmente. De tamaño y volumen variables según el ejemplar y la cantidad de líquido en su interior. La pared dorsal de la glándula vesicular en su parte craneal se relacionaba con la parte terminal del colon y recto. La parte caudal estaba en relación con la propróstata y en su extremo distal formaba el conducto eyaculatorio. La pared ventral se relacionaba con las ampollas del conducto deferente. El estudio histoquímico mostró que la glándula vesicular estaba constituida por una capa muscular con fibras musculares lisas organizadas irregularmente y una capa mucosa con una lámina propia rica en fibras colágenas y vasos sanguíneos, sobre la cual descansaba el epitelio glandular, simple, columnar bajo a cúbico. La histoquímica reveló la presencia de mucinas ácidas sulfatadas y no sulfatadas principalmente en la zona apical de las células glandulares y hacia el lumen de cada glándula. No fueron detectados glucógeno ni mucinas neutras.

Stereology and ultrastructure of the seminal vesicle of C57/BL/6J mice following chronic alcohol ingestion.

Excessive alcohol consumption causes metabolic changes and pathologic alterations in testes and accessory sex organ in different animal species. The aim of the present study was to evaluate the macroscopic, histologic and ultrastructural alterations provoked by chronic ingestion of different ethanol concentrations over increasing periods of time on the secretory epithelium of the seminal vesicle of C57/BL/6J mice in using stereological methods. Sixty male adult mice were divided into three experimental groups: Control, Alcoholic 25% and Alcoholic 35%, respectively, receiving tap water and tap water containing ethanol diluted to 25 and 35 degrees Gay Lussac. All mice were fed with the same solid diet. After 150 and 250 days of treatment the animals were sacrificed and the seminal vesicles were collected and processed for light and transmission electron microscopy. The cellular, cytoplasmic and nuclear volumes and the area density of autophagic and secretory vacuoles were measured. The histologic alterations observed in the alcoholic mice consisted of a reduction in epithelial size and cell volume, with maintenance of the same nuclear and cytoplasmic ratio as verified in the control groups. The ultrastructural alterations were: increased density of dense body area, decreased density of secretory granule area, and dilated rough endoplasmic reticulum and Golgi cisternae. We conclude that chronic ethanol ingestion causes depleting morphologic alterations in the epithelial cells of the seminal vesicle and negatively affects the secretory process of this gland.

Relationship between expression of sex steroid receptors and structure of the seminal vesicles after neonatal treatment of rats with potent or weak estrogens.

In this study we evaluated the effect of manipulating the estrogen and androgen environment of the neonatal male rat on subsequent immunoexpression of sex steroid receptors in the seminal vesicles (SVs) at age 18 days. The aim was to establish to what extent such changes were associated with and predictive of changes in SV structure/composition. Treatments were either diethylstilbestrol (DES; 10, 1, or 0.1 microg/injection), ethinyl estradiol (EE; 10 microg/injection), tamoxifen (2 mg/kg/day), flutamide (50 mg/kg), a gonadotropin-releasing hormone antagonist (GnRHa; 10 mg/kg), genistein (4 mg/kg/day), octylphenol (2 mg/injection), or bisphenol A (0.5 mg/injection). Compared with controls, treatment with DES (10 microg) induced loss of epithelial and stromal androgen receptor (AR) immunoexpression coincident with induction of stromal progesterone receptor (PR) immunoexpression and upregulation of stromal immunoexpression of estrogen receptor-alpha (ERalpha). These changes were associated with gross distortion (increase) of the normal stromal:epithelial tissue proportions in the SVs. DES (1 microg) and EE induced similar but less pronounced changes, and DES (0.1 microg) had no noticeable effect. Tamoxifen and flutamide induced PR and slightly upregulated ERalpha immunoexpression but had only a minor or no effect on AR expression and the stromal:epithelial ratio, though flutamide retarded normal development of the SVs. The latter was also evident in GnRHa-treated males, but otherwise this treatment had no effect on AR and PR immunoexpression. None of the foregoing treatments had any detectable effect on the immunoexpression of ERss in stromal or epithelial cells. The major treatment-induced changes in immunoexpression of AR, PR, and ERalpha and lack of change in ERss were confirmed by Western blots of SV protein extracts. None of the three weak (environmental) estrogens tested caused any detectable change in sex steroid receptor immunoexpression or SV tissue composition. We conclude that treatment-induced loss of AR is a prerequisite for altered stromal:epithelial proportions in the SVs and that such loss is always associated with induction of PR and upregulation of ERalpha; the latter two changes are insufficient on their own to bring about such a change. Nevertheless, induction of PR expression was always associated with altered SV development and is a potentially useful marker because it is not normally expressed in male reproductive tissues.

Modulation of prostaglandin H synthase activity by conjugated linoleic acid (CLA) and specific CLA isomers.

Conjugated linoleic acid (CLA) has been shown to inhibit tumorigenesis in animal models and is cytostatic to numerous cell lines in vitro. However, the mechanism of action is unknown. In the current study, we determined the effects of CLA and specific isomers of CLA on the rate of oxygenation of arachidonic acid by prostaglandin H synthase (PGHS) in ram seminal vesicle microsomes. The enzyme was incubated with 0.1 to 100 microM CLA or specific isomers of CLA for 2 min prior to the addition of 44 to 176 microM arachidonate. The isomers tested were 9(E),11(E) CLA; 9(Z),11(E) CLA; 9(Z),11(Z) CLA, and 10(E),12(2) CLA. For a positive inhibitor control, flurbiprofen was used at 0.75 to 2.50 microM. Enzyme activity was assessed by measuring the rate of oxygen consumption. Inclusion of CLA or specific isomers of CLA in the incubation mixtures inhibits PGHS. The efficacy differs for each isomer, with the 9(Z),11 (E) CLA isomer being the most effective and the 9(Z),11 (Z) CLA isomer being the least effective inhibitor among the four CLA isomers tested. The Ki values obtained by Dixon replots range from 18.7 microM for the most effective isomer, 9(Z),11 (E) CLA, to 105.3 microM for the least effective isomer, 9(2),11(2) CLA. The Ki value for flurbiprofen with ram seminal vesicle microsomes was 0.33 microM. As the concentration of arachidonate was increased, the CLA-dependent inhibition of PGHS decreased, suggesting competitive inhibition. The results of this study demonstrate the potential of CLA and specific isomers of CLA to modulate prostaglandin biosynthesis.

Ultrastructure of the seminal vesicle of Phlebotomus perniciosus Newstead (Diptera, Psychodidae).

The seminal vesicles of Phlebotomus perniciosus were investigated by light microscopy, confocal scanning laser microscopy and by scanning and transmission electron microscopy. They have a complex structure, and three different morphological compartments called A, B and C are distinguished on the basis of their position and fine structure. Compartment A is continuous with the vasa deferentia and consists of a cylindrical wall limiting a lumen in which the spermatozoa are stored. Compartment B is hemispherical and surrounds compartment A like a muff. Compartment C constitutes an external coat surrounding A and B. The epithelial cells of each compartment are characterized by morphologically different secretory granules. The ultrastructural features of these cells are described and their role in sandfly reproductive biology is discussed.