Prenatal differentiation of bovine oviductal epithelium: an electron microscopic study

In this study, we investigated the ultrastructural changes during the prenatal differentiation of oviductal epithelium in 16 bovine embryos and fetuses from CRL of 18.0 cm to a CRL of 94.0 cm. Ciliated and secretory cells of bovine uterine tube, a derivative of the Müllerian duct, differentiate to distinct development stages in the prenatal period. The typical cellular pattern, which is generally characteristic for the adult bovine oviduct, is also obtained during fetal life. In the early stages (CRL 18.0/20.4 cm), the bovine oviductal epithelium appears mostly undifferentiated. The epithelial cells show only a few mitochondria, some cisternae of rough endoplasmic reticulum (rER) and a small Golgi-complex. Most of the cytoplasm is filled with a large amount of glycogen, which decreases during later development. Interspersed between the undifferentiated epithelial cells, a few cells undergoing ciliogenesis can be observed. Ciliogenesis increased significantly during the later prenatal developmental stages. At a CRL of 55.0 cm, ciliated cells appear fully differentiated with mature cells covering their luminal surface. Formation of cilia usually use the acentriolar pathway. Fibrous granules occurred initially in association with the Golgi-apparatus and r(ER) in the supranuclear cytoplasm. Fibrous granules later fuse with deuterosomes and give rise to procentrioles, which are translocated to the luminal plasma membrane. There they become arranged in a line just beneath the apical cell membrane and further differentiate to basal bodies from which the formation of cilia and striated rootlets take place. Clear signs of differentiation of secretory cells were first seen in our material in fetuses with a CRL of 51.0 cm and 64.0 cm. These cells contain a well developed rER and Golgi-apparatus with dilated cisterns. In the supranuclear cytoplasm, the number of secretory granules continuously increases during later development and the cells adapt to the morphology of mature secretory cells at the CRL 94.0 cm.     

Expression and distribution of intermediate-filament proteins and laminin during the development of the bovine Müllerian duct

The expression pattern of several intermediate-filament proteins (vimentin, cytokeratin 8, 18 and 19) and the basal lamina component laminin was investigated in the Wolffian and the Müllerian ducts of bovine embryos and fetuses. The material studied comprised sexually undifferentiated stages [crown-rump length (CRL) 0.9 cm/1.0 cm/1.2 cm/1.9 cm/2.5 cm] and female stages (CRL 3.0 cm/4.2 cm/5.1 cm). Laminin could be demonstrated in the basal lamina of the developing Wolffian and Müllerian duct as well as in the stroma surrounding the Müllerian duct. The intermediate-filament protein vimentin was expressed in the mesothelium of the funnel field and in the epithelium of the Müllerian duct in all studied specimens, whereas the epithelial cells of the Wolffian duct only showed vimentin expression from a CRL of 2.2 cm onwards. In the cranial part of the Müllerian ducts only a few cells stained with pan-cytokeratin antibodies, whereas mesothelium and epithelium of the Wolffian duct showed as distinct immunostaining in all investigated stages. Both genital ducts showed no immunostaining with the antibody against cytokeratin 19 at any time of development. We conclude from our immunohistochemical results that the epithelial cells of the Wollfian duct do not contribute cells to the developing Müllerian duct.     

Functional indispensability of fibronectin associated on the mesenchymal cell surface during the early period of feather development in the chick embryo in vitro

To clarify the role of fibronectin (FN) during the early period of feather development, reconstituted skin consisting of intact epithelium and isolated mesenchymal cells from embryonal chick skin was used. In early feather development, FN was localized around mesenchymal cells of the dermal condensation. Isolated mesenchymal cells had associated with FN on their surfaces. FN on the cell surface dissociated following EDTA treatment, and EDTA‐treated cells re‐associated with exogenous FN. The intact epithelium also bound to exogenous FN at the placode. When FN‐associated or FN‐reassociated mesenchymal cells were used, the reconstituted skin formed feather rudiments only at the positions where the epithelial placode existed originally, and the locality of tenascin transferred from the placode to the mesenchyme during the period of feather bud formation. However, in reconstituted skin using FN‐dissociated mesenchymal cells, feather rudiments did not form. Additionally, the epithelial placodes disappeared, and tenascin was distributed uniformly on the surface of the epithelium and not localized in the mesenchyme. These findings suggest that FN associated on the surfaces of mesenchymal cells maintains the functions of mesenchymal cells as dermal condensation and mediates epithelial‐mesenchymal interactions during the early period of feather development. The results also suggest that reconstituted skin is a useful tool for functional studies on the extracellular matrix.     

Dermal-epidermal relationships in the skin of the bottlenose dolphin (Tursiops truncatus)

Using a variety of methods the outer surface of the dolphin dermis presents as a rather regularly arranged series of mainly longitudinal and approximately parallel ridges which are surmounted by laterally flattened, regularly distributed dermal papillae. At variable intervals groups of dermal ridges are supported by shallow elevations referred to here as basal dermal elevations. These tend to contain vessels and/or nerves for distribution to two or more dermal ridges. The opposing epidermal surface reveals an irregularly edged series of nearly parallel epidermal ridges alternating with the grooves which in the living state contain the dermal ridges. In the depths of these grooves are the crypts which in the intact skin contain the dermal papillae. Nothing is observable which could correspond to the epidermal pegs referred to in the literature. Small irregular epidermal projections (knobs) appear along the free edge of the epidermal ridge. Topographical variations in density and height of dermal ridges, height of basal dermal elevation plus dermal ridge and papillae as well as total epidermal thickness are illustrated by means of skin maps. Inconsistencies in the literature on Cetacean skin are discussed.     

Prenatal development of the bovine oviduct

In this study the development of the bovine Fallopian tube was investigated using light microscopic methods. Formation and differentiation of the Müllerian duct were studied in mesonephroi of 16 embryos and fetuses with a crown-rump lengths (CRL) of 0.9-8.4 cm. The funnel field, the rostral beginning of the Müllerian duct was first observed at a CRL of 0.9 cm. It appears as a thickening of the mesothelium on the craniolateral side of the mesonephros. During later development the Müllerian duct emerges by caudal outgrowth from the funnel field. Formation of a common basal lamina surrounding the caudal tips of Müllerian and Wolffian ducts could be observed at all stages up to CRL of 2.7 cm. The mesothelium and the epithelium of the Wolffian duct adjacent to the Müllerian duct showed a modification of epithelium height in all examined stages. Probably the Wolffian duct influences the growth of Müllerian duct by epithelio-mesenchymal interactions. Fetuses from a CRL of 12.0 to 94.0 cm were used for investigation of the prenatal differentiation of the oviductal mucosa. Folding of the oviductal mucosa started at a CRL of 29.0 cm and continued until birth. Individual primary, secondary and tertiary folds are formed in special proliferation zones and epithelium-folding buds. The cellular differentiation of the oviductal epithelium involves the formation of ciliated and secretory cells during different times of prenatal development. Ciliogenesis was first detected at a CRL of 33.0 cm. Active secretory cells could be observed in the oviductal epithelium from a CRL of 64.0 cm onwards.     

Development and cell differentiation of the motor nucleus of the facial nerve of cattle]

The early development, cell-migration and cell-differentiation of the nucleus motorius nervi facialis were studied in 32 bovine embryos with a CRL of 1 to 53 cm by light microscopical techniques. The ventro-medial cell column, a transitory embryonic formation, can be regarded as the origin of the nucleus. From there migrating cells can be demonstrated up to a CRL of 2.7 cm. With 3.8 cm CRL the cells are confined to their definitive location. From 5 cm CRL onwards a subdivision into 4 subnuclei can be seen. By succeeding maturation processes the nucleus of fetuses with 53 CRL acquires the topographical and cellular appearance of mature animals. With the electron microscope the cell-differentiation of the early stages (2.5 and 3.6 cm CRL) was demonstrated. Additionally the ventro-medial cell column was studied. The vertical columnar organisation of the neurons of the nucleus facialis shows besides longitudinal orientated guiding structures the migration process which is taken place at a CRL of 2.5 cm. Synaptogenic cell contact are seen from 3.6 cm SSL. At this stage the migration of cells has come to an end.     

Early development and cell differentiation of the parasympathetic vagus-glossopharyngeal nucleus in cattle. Light and electron microscopic studies]

The early development, differentiation of the cell and cell migration of the nucleus parasympathicus nervi vagi et glossopharyngei were examined by light microscope in 32 bovine embryos with crown-rump-lengths (CRL) ranging from 1 cm to 53 cm. During this period the nucleus is being enlarged 6 to 7 times and the size of the cell increases to 35-40 microns. The ultrastructure during the differentiation of the cell is shown electron microscopically in embryos with CRL of 2.5 cm and 3.6 cm. Several layers of matrix cells arise from the neuroepithelium of the neural tube by mitosis. They migrate in the shape of dark nucleated cells into the parasympathetic cell column. With advancing age of the embryos the number of cells with light nucleus increases. They represent the presumptive neurons. In embryos of 2.5 cm CRL their cytoplasm surrounds the nucleus on three sides in the shape of a narrow rim while on the fourth side it is enlarged into an outgrowing process. In this process a smaller number of organelles and their preliminary stages appears. Their number is significantly increased in embryos of 3.6 cm CRL and they can be seen throughout the growing process. In the following stages of maturity cytological development proceeds. In embryos of 53 cm CRL topographical and cytological data are comparable to those in adult animals.     

Morpho- und Histogenese der Colliculi rostrales beim Rind

Based upon macroscopic and light microscopic examinations the development of the superior colliculus of bovine embryos and fetuses with crown-rump lengths (CRL) ranging from 0.8 to 90.0 cm was studied. Macroscopically, the mesencephalon can be recognized for the first time at 0.8 cm CRL, whereas the superior colliculus can be clearly differentiated in embryos of 4.5 cm CRL. The macroscopic features of this brain area have reached adult conditions at 80.0 cm CRL. The light microscopic examination reflected the beginning of layer formation at 0.8 cm CRL, which is induced by the proliferating activity of the ventricular zone. Up to 3.4 cm CRL the primordium of the tectum opticum exhibits still a trilaminate pattern (ventricular-, intermediate- and marginal zone), but at 4.5 cm CRL, the formation of the specific tectal layers is marked by the origin of the stratum profundum and intermedium. With the appearance of the tenth layer named stratum opticum at 8.0 cm CRL the laminar pattern corresponds to the characteristics of adult animals.     

Localization and expression of gastrin-releasing peptide (GRP) in the bovine cervix

Gastrin-releasing peptide (GRP) has been suggested as a novel regulatory peptide in the female reproductive tract but the presence of GRP and GRP mRNA in the non-neurogenic tissue of the cervix has not yet been clarified. In the present study, immunohistochemistry and in situ hybridization were used to reveal the distribution of GRP immunoreactivity and expression of GRP mRNA in the bovine cervix. The cervixes from 21 non-pregnant and 20 pregnant cows, and 6 fetuses were used in the study. In the fetus, adult non-pregnant and pregnant specimens, GRP and GRP mRNA were predominantly detected in the luminal epithelial cells of basal areas of peripheral regions of the cervix. Positive staining of GRP in the epithelial cells of the cervix was first detected in the CRL 37 cm of the fetus. During the estrous cycles, the staining intensity of GRP in the epithelial cells was stronger in the follicular phase than in the luteal phase. During the early gestational period, GRP immunoreactivity was detected at relatively similar intensity to the follicular phase. In situ hybridization results ascertained the expression of GRP mRNA in the superficial epithelial cells of the cervix of non-pregnant and pregnant cows. The results suggest that GRP may be important both in the development of the fetal cervix and secretory activity of the epithelial cells of the cervix.     

Stereological assessment of the epithelial surface area of the ovine palatine and pharyngeal tonsils

Although ovine tonsils play a major role in prion diseases, their morphology is poorly documented and morphometric data are sparse. Therefore, a stereological assessment of the surface area of the palatine and pharyngeal tonsils of five sheep was performed. The epithelial surface area of both tonsils is considerably increased by the presence of tonsillar crypts and folds. The mean epithelial surface area of the paired palatine tonsil was 7.14 cm(2) and that of the pharyngeal tonsil was 23.5 cm(2). In comparison with the paired palatine tonsil, the volume of the pharyngeal tonsil was almost two times larger, whilst its surface area was more than three times larger.     

Carbonic Anhydrase III in the Salivary Glands and Kidney of the Japanes Monkey (Macaca fuscata)

Carbonic anhydrase III (CA-III) was found in muscles of the Japanese monkey by the double immunodiffusion test and western blotting using antiserum raised against equine CA-III. Immunocytochemical localization of CA-III in the salivary glands and kidney of the monkey was studied using an avidin-biotinylated glucose oxidase complex. CA-III was found mainly in the striated duct and interlobular duct cells of the parotid glands. In the submandibular glands, striated duct, interlobular duct, and excretory duct cells were strongly stained for CA-III. In the kidney of the monkey, CA-III was localized mainly in the dark cells of the collecting duct at the medulla and in the epithelial cells of thick limb of Henle's loop.     

Studies on the Ovarian Development in Zebu Cattle (Bos indicus)

In a study of 71 female foetuses, gonadal blastema was observed at 1.5 cm crown rump length (CRL). Oogonial cells entered the meiotic prophase at 3.5-6.0 cm CRL, which was arrested at the dictyotene stage to produce primary oocytes which formed primordial follicles. Primordial follicles were observed at 6-8 cm CRL. All germinal cells were at dictyotene by 20-24 cm CRL and follicles developed to primary and secondary stages. Folliculogenesis dominated further ovarian development and reached a peak between 32 and 35 cm CRL. In seven of the 12 foetuses measuring between 41 and 72 cm CRL, many follicles were atretic and some luteinized. The luteal bodies were composed of hypertrophied theca and granulosa cells with homogeneous and eosinophilic cytoplasm.     

沟垄集雨对紫花苜蓿和裸燕麦出苗及土壤贮水量的影响

采用沟垄覆膜集雨种植紫花苜蓿和裸燕麦,测定不同沟垄比和不同垄覆盖材料对植物土壤贮水量和出苗数的影响。结果表明,沟中土壤贮水量随沟垄比增加而增加。普通地膜、可降解地膜和自然降水形成的土壤结皮具有减少土壤蒸发的作用,从而提高土壤贮水量。2012年5月7日紫花苜蓿出苗期测定结果显示,普通膜垄、可降解膜垄和土垄种植紫花苜蓿的土壤贮水量比平作分别提高为77,61和38mm。2012年4月27日裸燕麦出苗期测定结果显示,普通膜垄、可降解膜垄和土垄种植裸燕麦的土壤贮水量比平作分别提高102,83和61mm。普通地膜、可降解地膜和土垄处理提高了土壤含水量,从而提高裸燕麦株数;普通膜垄、可降解膜垄和土垄的裸燕麦株数比平作分别提高47,37和11株/m2。但高强度集雨不一定利于紫花苜蓿发芽,平作土壤含水量分布均匀,更有利于紫花苜蓿发芽。     

On the development of the papillary body in the feline claw

The pre- and post-natal development of the feline claw was studied in 22 feline fetuses with a crown-rump length (CRL) ranging from 40 to 160 mm, in six kittens up to an age of 22 days, and in four adult cats. In fetuses up to a CRL of 75 mm, the characteristic shape of the feline claw was developed. Segment-specific dermal modifications in the various segments, especially the dorsal ridge, started to develop in fetuses with a CRL between 75 and 105 mm. Modifications of the papillary body in the different claw segments took place in the last third of prenatal development and were continued postnatally. At first, the basal lamina became wavy, followed by the formation of small dermal microridges, which would be enlarged to dermal ridges and lamellae. In the claw of adult cats, the papillary body was very small. The dermal tissue of the proximal part of the coronet formed low microridges with short papillae originating on and between these low ridges. In the wall segment, dermal microridges were formed which were arranged in a parallel fashion, and in the distal part of the wall, short dermal micropapillae arose on the crest of each microridge. In the sole segment, thin dermal lamellae were developed. The sequence of papillary body development and the varying conformations of the papillary body in the different segments of the feline claw are compared to those in the nail, the canine claw and hooves.     

Isolation and measurement of carbonic anhydrase isoenzyme III in plasma, sera, and tissues of dogs

OBJECTIVE: To purify canine carbonic anhydrase isoenzyme III (CA-III) and determine plasma, serum, and tissue concentrations of CA-III in healthy dogs and dogs with experimentally induced muscle damage. ANIMALS: 121 healthy Beagles. PROCEDURE: Muscle was obtained from 2 Beagles after euthanasia, and CA-III was purified and characterized by use of column chromatography and electrophoresis, respectively. A CA-III-specific ELISA was developed to determine concentrations of CA-III in plasma of 116 dogs and tissues of 1 dog. Serum creatine kinase (CK) activity and CA-III concentration were also determined before and after induction of muscle damage by IM injection of 2 ml of 10% lidocaine to 2 dogs. RESULTS: Canine CA-III had a molecular weight of 28 kd and an isoelectric point of 8.2. Mean (+/- SD) concentration of CA-III in plasma of healthy dogs was 16.91 +/- 9.55 ng/ml. The highest tissue concentration of CA-III was detected in skeletal muscle. Serum concentration of CA-III increased and peaked within the first 2 to 3 hours after induction of muscle damage. The increase in CA-III concentration was more rapid than that of CK activity, and concentration reached its maximum and returned to baseline sooner than did CK activity. CONCLUSIONS AND CLINICAL RELEVANCE: The CA-III ELISA we developed was a sensitive method for determining CA-III concentrations in plasma, serum samples, and tissue specimens of dogs. Use of this ELISA requires only a small volume of serum and may enable the study of changes in CA isoenzyme concentrations associated with muscle disorders in dogs.     

Role of beta1 integrins in adhesion of canine mastocytoma cells to extracellular matrix proteins

The interactions of tumor cells with the extracellular matrix (ECM) are a crucial step in invasion and metastasis. Integrins are adhesive molecules forming heterodimers with alpha and beta subunits that play a definitive role in these interactions. In this study, mastocytoma (mast cell tumor: MCT) cell-ECM interaction was investigated using 3 canine MCT cell lines: CM-MC (originating from cutaneous MCT), VI-MC (originating from intestinal MCT), and CoMS (originating from oral MCT). Flow cytometric analysis showed that all cells highly expressed the integrin beta1 and alpha1 through alpha5 subunits and that they moderately expressed the alpha6 subunit. In adhesion studies, CoMS weakly but spontaneously adhered to fibronectin (FN), which was enhanced by phorbol ester (TPA), while CM-MC and VI-MC required cell activation by TPA to adhere to FN. Anti-beta1 and alpha5 integrin antibodies strongly inhibited cell adhesion to FN in CM-MC and CoMS and moderately inhibited cell adhesion in VI-MC. Only VI-MC adhered to laminin (LN) under activation by TPA. Anti-beta1 integrin antibodies strongly inhibited cell adhesion to LN, but all anti-alpha integrin antibodies failed to inhibit cell adhesion to LN. No cells adhered to collagen types I and IV. Canine MCT cells from different origins expressed similar integrin patterns; however, there were some differences in adhesive behavior in response to various ECM proteins and activating stimuli.     

Entwicklung,Zytoarchitektonik und Feinstruktur des mesencephalen Trigeminuskerns bei Hauswiederkäuern *

Development, cytoarchitecture and ultrastructure of the mesencephalic trigeminal nucleus in domestic ruminants The ontogenetic development and cell differentiation of the mesencephalic trigeminal nucleus (Ntm) is lightmicroscopically examined in 58 bovine embryos and fetuses ranging from 2.4 to 80 cm Crown-Rump-Length (CRL). The cytoarchitecture and fine structure in adult cattle, sheep, and goats are investigated with the aid of light- and electronmicroscopy. At 2.4 cm CRL, the proneurons of the Ntm are detectable for the first time within the ventricular zone of the alar plate, possessing one drop-like cytoplasmic protrusion, whereas at 5 cm CRL, two cell types with differing sizes appear. Up to a CRL of 11.5cm, the nucleus shows advanced maturation processes and has reached his final position at the border of the mesencephalic central grey. From 26 cm CRL onward, three cell types, and at 34 cm CRL four cell types, are discernible based on their nissl-granule arrangement. The cytomorphological differentiation and the maturation of the cells proceeds until 56 cm CRL, at which point the topographical and cytological characteristics of the Ntm are comparable with those of adult animals. In adult cattle, sheep and goats the Ntm consists of large (40–60 μm) and scarce medium-sized (30–40μm) neurons with round and oval shapes. Scarcer small (20–25 μm) round and medium-sized multipolar neurons occur. The Nissl bodies are scattered throughout the pericaryon of the large neurons in a dust-like pattern and in the medium-sized neurons in a grained form. Within the cytoplasmic streets, which are situated between the membranes of the rough ER, numerous neurofilaments and mitochondria are detectable. Large Golgi complexes are placed in a perinuclear position. The neurons are also characterized by some somatic spines, and by a moderate distribution of axosomatic synapses, in which axon-endings with flattened synaptic vesicles predominate.     

Immunocytochemical localisation of carbonic anhydrase isozyme III in equine skeletal muscle

The location of carbonic anhydrase III (CA-III) in frozen sections of biopsies of Thoroughbred horse skeletal muscle was studied. Fibre types were determined by ATP-ase and succinate dehydrogenase staining. CA-III isozyme was detected using a peroxidase conjugated anti-CA-III antibody. CA-III was found to be localised in slow twitch oxidative fibres (ST), but was also present in fast twitch oxidative (FTH) fibres in small amounts. Fast twitch glycolytic (FT) fibres were stained lightly compared with control sections. The concentrations of CA-III in muscle and liver were 70 micrograms/mg protein and 4 micrograms/mg protein, respectively. CA-I and CA-II were not found in muscle extracts by the double immunodiffusion method.