Licht- und elektronenmikroskopische Untersuchungen zur prä- und postnatalen Entwicklung des Eileiters beim Rind

Anhand von histologischen, immun- und lektinhistochemischen Untersuchungen wurde die Entwicklung des Eileiters beim Rind analysiert. Hierbei wurden sowohl die Bildung und Differenzierung des Müller Ganges als auch des pränatalen Eileiters untersucht

Fetal Development of the Bovine Uterus: A Light Microscopy and Immunohistochemical Study

Important steps during the prenatal development of the bovine uterus are described using conventional hematoxylin-eosin staining of fetuses from different developmental stages [crown-rump length (CRL) 9.2-94.0 cm]. Additionally, a number of intermediate filaments (keratin 7, 8, 14, 18, 19; and vimentin), the basement membrane protein laminin, smooth-muscle marker (SMA), and S100 were studied to further characterize certain differentiation processes. During early development, the uterine epithelium is simple or (pseudo)stratified with bud-like protrusions. Developing caruncles can be observed in the corpus uteri at a CRL of 15.8 cm onwards, showing a simple, keratin-positive epithelium. In contrast, the intercaruncular areas are characterized by a (pseudo)stratified epithelium, which also shows positive staining in a different manner for the investigated keratins. A differentiation of smooth muscle cell layers can be observed from a CRL of 24.4 cm onwards. Intense SMA-positive cells/fibers, arranged perpendicularly to the developing circular SMA-positive muscle cell layer, can be found preferentially located in the developing caruncles. Lymphocytes occur in the uterine epithelium and stroma in the corpora and cornua of fetuses with a CLR of 15.8 cm and higher. (C) 2016 S. Karger AG, Base

Fetal Development of the Bovine Uterus: A Light Microscopy and Immunohistochemical Study

Important steps during the prenatal development of the bovine uterus are described using conventional hematoxylin-eosin staining of fetuses from different developmental stages [crown-rump length (CRL) 9.2-94.0 cm]. Additionally, a number of intermediate filaments (keratin 7, 8, 14, 18, 19; and vimentin), the basement membrane protein laminin, smooth-muscle marker (SMA), and S100 were studied to further characterize certain differentiation processes. During early development, the uterine epithelium is simple or (pseudo)stratified with bud-like protrusions. Developing caruncles can be observed in the corpus uteri at a CRL of 15.8 cm onwards, showing a simple, keratin-positive epithelium. In contrast, the intercaruncular areas are characterized by a (pseudo)stratified epithelium, which also shows positive staining in a different manner for the investigated keratins. A differentiation of smooth muscle cell layers can be observed from a CRL of 24.4 cm onwards. Intense SMA-positive cells/fibers, arranged perpendicularly to the developing circular SMA-positive muscle cell layer, can be found preferentially located in the developing caruncles. Lymphocytes occur in the uterine epithelium and stroma in the corpora and cornua of fetuses with a CLR of 15.8 cm and higher. (C) 2016 S. Karger AG, Base

Expression of Intermediate Filaments and Germ Cell Markers in the Developing Bovine Ovary: An Immunohistochemical and Laser-Assisted Microdissection Study

In the present investigation, bovine ovary prenatal development was studied using immunohistochemistry and laser-assisted microdissection (LAM). A major aim of this study was to evaluate the protein expression pattern of intermediate filaments (IF) and distinguish S100 protein (S100 alpha and S100 beta protein) isoforms during prenatal follicle differentiation, subsequently correlating them with germ cell marker expression. A development-specific expression pattern of different keratins as well as vimentin was detected in the prenatal bovine ovary; K18-specific expression was found during all developmental stages (i.e. in surface epithelium, germ cell cord somatic cells, and follicle cells), and keratins 5, 7, 8, 14, and 19 and vimentin had a stage-specific expression pattern in the different cell populations of the prenatal ovaries. Additionally, our results represent new data on the expression pattern of germ cell markers during bovine ovary prenatal development. S100 alpha and beta protein was localized to oocyte cytoplasm of different follicle stages, and S100 alpha staining could be observed in granulosa cells. Furthermore, through isolation of characteristic ovary cell populations using LAM, specific confirmation of some genes of interest (KRT8, KRT18, S100 alpha, S100 beta, and OCT4, DDX4) could be obtained by RT-PCR in single cell groups of the developing bovine ovary.© 2015 S. Karger AG, Base

Expression of Intermediate Filaments and Germ Cell Markers in the Developing Bovine Ovary: An Immunohistochemical and Laser-Assisted Microdissection Study

In the present investigation, bovine ovary prenatal development was studied using immunohistochemistry and laser-assisted microdissection (LAM). A major aim of this study was to evaluate the protein expression pattern of intermediate filaments (IF) and distinguish S100 protein (S100 alpha and S100 beta protein) isoforms during prenatal follicle differentiation, subsequently correlating them with germ cell marker expression. A development-specific expression pattern of different keratins as well as vimentin was detected in the prenatal bovine ovary; K18-specific expression was found during all developmental stages (i.e. in surface epithelium, germ cell cord somatic cells, and follicle cells), and keratins 5, 7, 8, 14, and 19 and vimentin had a stage-specific expression pattern in the different cell populations of the prenatal ovaries. Additionally, our results represent new data on the expression pattern of germ cell markers during bovine ovary prenatal development. S100 alpha and beta protein was localized to oocyte cytoplasm of different follicle stages, and S100 alpha staining could be observed in granulosa cells. Furthermore, through isolation of characteristic ovary cell populations using LAM, specific confirmation of some genes of interest (KRT8, KRT18, S100 alpha, S100 beta, and OCT4, DDX4) could be obtained by RT-PCR in single cell groups of the developing bovine ovary.© 2015 S. Karger AG, Base

Localization of gene and protein expressions of tumor necrosis factor-alpha and tumor necrosis factor receptor types I and II in the bovine corpus luteum during the estrous cycle

One of the many roles of tumor necrosis factor (TNF)-α is to control mammalian corpus luteum (CL) PG synthesis and apoptotic cell death. Here, the cellular localization of TNF-α and its type I (TNF-RI) and type II (TNF-RII) receptors in bovine luteal tissue were analyzed using in situ hybridization, immunohistochemistry, and quantitative real-time PCR. Transcripts for TNF-α were expressed in bovine CL throughout the estrous cycle, but were significantly more abundant (P < 0.01) at the regressed luteal stage than at the other stages. Localization of TNF-α transcripts and protein were observed in large and small bovine luteal cells, as well as in immune cells. Moreover, transcripts for TNF-RI and TNF-RII were expressed in bovine CL throughout the estrous cycle. The abundance of TNF-RII transcripts was greater (P < 0.01) at the regressed luteal stage than at the other stages, whereas TNF-RI transcript abundance did not significantly change. Expression of TNF-RI and TNF-RII transcripts and proteins were observed in both the large and small luteal cells, and the proteins were also expressed in the immune cells and vascular endothelial cells. These results suggest that TNF-α sources include immune cells, as well as large and small luteal cells, and that TNF-RI and TNF-RII are present in the luteal cells of the bovine CL

Localization of gene and protein expressions of tumor necrosis factor-alpha and tumor necrosis factor receptor types I and II in the bovine corpus luteum during the estrous cycle

One of the many roles of tumor necrosis factor (TNF)-α is to control mammalian corpus luteum (CL) PG synthesis and apoptotic cell death. Here, the cellular localization of TNF-α and its type I (TNF-RI) and type II (TNF-RII) receptors in bovine luteal tissue were analyzed using in situ hybridization, immunohistochemistry, and quantitative real-time PCR. Transcripts for TNF-α were expressed in bovine CL throughout the estrous cycle, but were significantly more abundant (P < 0.01) at the regressed luteal stage than at the other stages. Localization of TNF-α transcripts and protein were observed in large and small bovine luteal cells, as well as in immune cells. Moreover, transcripts for TNF-RI and TNF-RII were expressed in bovine CL throughout the estrous cycle. The abundance of TNF-RII transcripts was greater (P < 0.01) at the regressed luteal stage than at the other stages, whereas TNF-RI transcript abundance did not significantly change. Expression of TNF-RI and TNF-RII transcripts and proteins were observed in both the large and small luteal cells, and the proteins were also expressed in the immune cells and vascular endothelial cells. These results suggest that TNF-α sources include immune cells, as well as large and small luteal cells, and that TNF-RI and TNF-RII are present in the luteal cells of the bovine CL

The Role of Ovarian Surface Epithelium in Folliculogenesis during Fetal Development of the Bovine Ovary: A Histological and Immunohistochemical Study

Although many aspects of ovarian differentiation have been established, comparatively little is known about prenatal follicle formation and differentiation of bovine ovaries. The objective of this investigation was to study the role of the surface epithelium during the development of germ cell nests, germ cell cords and follicle formation in the fetal bovine ovary. Associated important proliferation and apoptotic features were further investigated. Additionally, the expression pattern of the S100 protein was detected. A strong increase of mitotic figures was detected in the surface epithelium, germ cell nests and germ cell cords of ovaries with a crown-rump length (CRL) of 13.0-58.0 cm. Oocytes were positively stained with S100 in bovine ovaries from fetuses with a CRL of 21.0 cm. The staining intensity enhanced parallel to increasing oocyte and follicle sizes during the ovary development. In later stages, a strong staining for S100 was observed in healthy oocytes in contradistinction to atretic oocytes where no expression of the S100 protein could be found. In conclusion, increasing mitosis index of surface epithelium cells, as well as oogonia directly beneath the surface epithelium, in combination with open surface connection during stages from a CRL of 11.0-94.0 cm of bovine fetal ovaries could play an important role in the period of time of ongoing folliculogenesis and derivation of granulosa cells. Additionally, S100-positive oocytes in primordial and later follicle stages joined by a high rate of Ki67-positive index in surrounding granulosa cells indicate that in the oocytes the S100 protein can perhaps be a useful marker for intact oocytes in bovine ovaries

In Vivo Biofilm Formation of Pathogenic Leptospira spp. in the Vitreous Humor of Horses with Recurrent Uveitis

Equine recurrent uveitis (ERU) causes painful inflammatory attacks and oftentimes blindness in the affected eyes. The disease is considered a late sequela of systemic leptospirosis. The most effective therapy is the surgical removal of the vitreous (vitrectomy), which is not only therapeutic, but provides vitreous material that can be assessed diagnostically. For example, the lipL32 gene, culturable Leptospira spp., and anti-Leptospira antibodies have all been detected in vitreous samples obtained from eyes with chronic ERU. Despite this clear evidence of leptospiral involvement, the systemic administration of antibiotics in infected horses is ineffective at resolving ERU. This syndrome of chronic recurrent inflammation, which is unresponsive to antibiotic therapy, combined with apparent bacteria evading the immune response, is consistent with a biofilm-associated infection. The purpose of this study, therefore, was to detect the in vivo biofilm formation of Leptospira spp. in vitreous samples collected during vitrectomy and examined using a Warthin-Starry silver stain and immunohistochemistry. All known steps of biofilm formation were visualized in these samples, including individual Leptospira spp., leptospiral microcolonies and dense roundish accumulations of Leptospira spp. In many instances spirochetes were surrounded by an extracellular substance. Taken together, data from the present study show that ERU is a biofilm-associated intraocular leptospiral infection, which best explains the typical clinical course

Histochemical Analysis of Glycoconjugates in the Skin of a Catfish (Arius Tenuispinis, Day)

A histochemical study using conventional carbohydrate histochemistry (periodic-acid staining including diastase controls, alcian blue staining at pH 1 and 2.5) as well as using a battery of 14 fluorescein isothiocyanate (FITC)-labelled lectins to identify glycoconjugates present in 10 different areas of the skin of a catfish (Arius tenuispinis) was carried out. The lectins used were: mannose-binding lectins (Con A, LCA and PSA), galactose-binding lectins (PNA, RCA), N-acetylgalactosamine-binding lectins (DBA, SBA, SJA and GSL I), N-acetylglucosamine-binding lectins (WGA and WGAs), fucose-binding lectins (UEA) and lectins which bind to complex carbohydrate configurations (PHA E, PHA L). Conventional glycoconjugate staining (PAS staining, alcian blue at pH 1 and 2.5) showed that the mucous goblet cells contain a considerable amount of glycoconjugates in all locations of the skin, whereas the other unicellular gland type, the club cells, lacked these glycoconjugates. The glycoproteins found in goblet cells are neutral and therefore stain magenta when subjected to PAS staining. Alcian blue staining indicating acid glycoproteins was distinctly positive at pH 1, but gave only a comparable staining at pH 2.5. The mucus of the goblet cells therefore also contains acid glycoproteins rich in sulphate groups. Using FITC-labelled lectins, the carbohydrate composition of the glycoproteins of goblet cells could be more fully characterized. A distinct staining of the mucus of goblet cells was found with the mannose-binding lectins LCA and PSA; the galactosamine-binding lectins DBA, SBA and GLS I; the glucosamine-binding lectin WGA; and PHA E which stains glycoproteins with complex carbohydrate configurations. No reaction occurred with the fucose-binding lectin UEA and the sialic acid-specific lectin SNA. In addition, the galactose-binding lectins PNA and RCA showed only a weak or completely negative staining of the mucus in the goblet cells. The specificity of the lectin staining could be proved by inhibiting binding of the lectins by competitive inhibition with the corresponding sugars. From these data, we can conclude that the mucus produced by the epidermal goblet cells of A. tenuispinis is rich in mannose, N-acetylgalactosamine and N-acetylglucosamine residues