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.     

Entwicklung und Zelldifferenzierung des Nucleus nervi hypoglossi beim Rind

Based upon light- and electron-microscope examinations, the ontogenetic development of the nucleus of cranial nerve XII is documented. At 1-cm crown-rump length (CRL), the caudal pole of the nucleus nervi hypoglossi forms a uniform cell column with the cornu ventrale of the spinal cord. During this period, its caudal area shows signs of cellular degeneration. From 3.5 cm CRL onward, all nuclear groups can be identified. At 53 cm CRL, they correspond to the pattern as described in the adult brain. Electron-optically, at 2.5 cm and 3.6 cm CRL, the nucleus of cranial nerve XII exhibits a close relationship to the matrix layer which consists of elements of dark nuclei. The hypoglossus nucleus is composed of dark and light cell types. It is the latter type that represents the presumptive neuron; it shows an increased ultrastructural differentiation from 2.5 cm CRL onward.     

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.     

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.     

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.     

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.     

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.     

Development of the glandular epithelium of the bovine parotid gland during ontogenesis

The development of the parotid gland was examined in 36 bovine embryos and foetuses with a crown-rump-length (CRL) from 28 up to 1000 mm by light, transmission electron microscopical and actin-immunohistochemical methods. The anlage of the parotid gland in an embryo with 28 mm CRL can be found at the lateral angle of the primitive oral cavity as a local thickening of the epithelium. During the second month, the differentiation of primary ducts and endbuds starts and a lumen develops in the primary ducts. At the end of the second month a lumen appears in the terminal endbuds. In the immature endpiece cells first secretory granules can be seen from a CRL of 240 mm. In the third month differentiation between intra- and inter-lobular ducts is possible. Immature myoepithelial cells present as a basal layer of flattened cells between the epithelial cells and the basement membrane at the end of the second month. During further development they increase in number, become more flattened and form long cellular processes. At the end of the fourth month isolated actin filament bundles are formed, which were also detected by an antibody against smooth muscle actin. The actin filaments condense continuously until they fill the cell processes completely at the end of foetal development.     

Die Entwicklung des Dottersackes beim Wiederkäuer (Schaf und Rind)

Yolk sac development was investigated in 69 ovine and 10 bovine embryos from the blastocyst stage to the 7th week of gestation. Light and electron microscopical findings are reported. The yolk sac in sheep and cattle is composed of an enlarged sac-like portion lying below the embryo and two ends which follow the elongated course of the trophoblast. In sheep, an open connection exists between the intestines and the yolk sac up to a crown-rump length (CRL) of 9 mm. It is closed by 12 mm CRL. The wall of the yolk sac is especially well vascularized in the enlarged, sac-like portion. Primary erythropoiesis occurs within the blood capillaries. In the blastocyst, the yolk sac entoderm is made up of elongated, flat cells. It becomes cuboidal in the 3 mm embryo (ovine) and later columnar. The up to 20 microns tall cells stain darkly and contain numerous light-colored vesicles. At 4.5 mm CRL light cells appear between the dark ones. Both cells are rich in rough endoplasmic reticulum (rER). The increased staining of the darker cells is due to an osmophilic cytoplasm and numerous, often parallel lamella of rER. The rER of the light cells is enlarged to irregularly-shaped cisternae, which nearly fill the entire cytoplasm and give them a rounded appearance. The dark cells contain polygonal nuclei, whereas those in the light cells are round with one or two nucleoli. The oval mitochondria have only a few peripheral cristae. Golgi fields are not very common. Cells of the entoderm are connected to one another over zonulae occludentes. They possess microvilli on the luminal surface and are supported by a basement membrane. From 5 mm CRL onwards (ovine), the yolk sac entoderm folds itself between the capillaries, thereby becoming stratified. The intercellular space between the cells expands as projections between neighboring cells interlock. Canaliculi arise between adjacent epithelia. The wall of the yolk sac thickens as a result of this infolding and the densely packed capillaries. Infoldings are especially predominant in the sac-like portion of the yolk sac, and only suggested in the ends. Involution of the yolk sac begins in the peripheral end segments and proceeds centripetally. Numerous glycogen particles appear in the yolk sac entoderm cells of the ovine fetus at a CRL of 36 mm, and by a CRL of 42 mm, the sac-like portion has also begun to show signs of degeneration. Mesenchyme is very sparse within the wall of the yolk sac throughout the entire period of development.(ABSTRACT TRUNCATED AT 400 WORDS)     

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.     

Distributions of extracellular matrix and carbonic anhydrase-III during bovine palatine ridge development.

The present study describes histological alterations and immunohistochemical distributions of extracellular matrices (ECMs) and the carbonic anhydrase isozyme-III (CA-III) during the period of bovine palatine ridge formation. Morphogenesis of bovine palatine ridges was preceded by epidermal placodes and the mesenchymal condensation (MC). During the early stages of less than 44 cm crown rump length (CRL), fibronectin (FN) was distributed densely in the MC. Strong reactions against type I collagen (C-1) were detected outer to the FN positive site. In the stages of more than 44 cm CRL, FN and C-1 were distributed diffusely in subepithelial mesenchyme. Laminin (LN) and type IV collagen were distributed in the epithelial and endothelial basement membranes (BMs) in all of the stages examined, except in the stage of 7 cm CRL, where LN was not detected only in the BM just beneath the epidermal placode. CA-III was detected in basal epithelial cells except for palatine ridge rudiments in the stages of more than 21 cm CRL. It is suggested that the expressions of LN and CA-III might play a role in the spatial determination of rudiments of bovine fetal palatine ridges.     

Quantification,Morphology and Ultrastructure of Preantral Follicles of Buffalo (Bubalus bubalis) Foetuses

The objective of this study was to determine the number, morphology and ultrastructure of preantral ovarian follicles of buffalo (Bubalus bubalis) foetuses at different ages. Quantification revealed number of primordial, primary and secondary follicles of 48 857 ± 17 506, 26 000 ± 20 452, 18 428 ± 10 875 and 18 375 ± 19 690, 225 ± 349, 326 ± 288 at 12–34 cm and 35–60 cm crown rump length (CRL), respectively. Follicular diameter values were 28.9 (±3.4), 34.7 (±5.9) and 59.4 (±12.6) μm; oocyte diameters were 21.7 (±2.8), 24.3 (±3.4) and 33.0 (±7.7) μm, and the numbers of follicular cells in the follicle equatorial section were 7.1 (±1.4), 12.0 (±2.4) and 13.8 (±2.4) for primordial, primary and secondary follicles, respectively. The primordial follicle consisted of an oocyte surrounded by a layer of flattened follicular cells with a normally eccentric oocyte nucleus. Dispersed Golgi complex, smooth endoplasmic reticulum, rounded mitochondria and several lipid vesicles were observed in the cytoplasm and cell junctions between the follicle cell membranes and the oocyte. This work describes the number, morphometry and ultrastructure of preantral follicles of buffalo foetuses, concluding that folliculogenesis is established between 8 and 34 cm CRL and that follicle number varies individually and according to age and that further studies are needed in this species.     

Ultrasonographic examinations of fetal growth of sheep between day 20 and day 50 of gestation

Previous studies on embryonic and fetal growth in sheep were mostly transversal using animals killed at various stages of gestation. Until now it was difficult to monitor the development of individual embryos/foetuses during pregnancy, especially during the first and second pregnancy month. Real-time ultrasound as a non invasive method could be an appropriate method for examination of embryonic and early foetal development in sheep. The aim of this study was to determine the embryonic and foetal development of the crown-rump-length (CRL) in pregnant ewes in relation to the number of fetuses and/or the breed. Between the 20th and 50th day of pregnancy the embryos/foetuses showed an exponential growth which can be best described by the equation of the form CRL (mm) = W * exp (k * day of pregnancy). The individual variability in embryofetal growth is in part due to the number of embryos per sheep and the sheep breed.     

B‐cell lymphoma with Mott cell differentiation in a cat

A 12‐year‐old, male castrated Domestic Shorthair cat was presented to Animal Medical Center of Gifu Univeristy with anorexia and vomiting. Physical examination revealed an enlarged left tonsil and right mandibular lymph node (approximately 2–3× the normal size), and a submucosal mass on the right side of the epiglottis (1.5 × 2.0 cm). On computed tomography images, an enlarged left tonsil, and enlarged right mandibular, right pharyngeal, and left and right cervical lymph nodes were observed. Cytologic examination of smears of tonsil and lymph nodes revealed numerous medium‐ to large‐sized neoplastic lymphoid cells, approximately half of which contained one or several light‐blue homogenous globoid cytoplasmic inclusions (5–10 μm), which stained magenta with periodic acid–Schiff (PAS) stain. Histopathologic examination of the left tonsil revealed diffuse proliferation of medium‐ to large‐sized neoplastic lymphoid cells effacing the original lymphoid architecture. Half of the cells contained one or several eosinophilic globoid cytoplasmic inclusions, which stained magenta with PAS and showed positive immunohistochemical reactions for immunoglobulin M (IgM) and λ light chain. Neoplastic lymphoid cells were also CD20⁺, Pax5⁺, and MUM1⁺, and CD3^?. Thus, the neoplastic lymphoid cells expressed a B‐cell immunophenotype, and the globoid cytoplasmic inclusions represented an aberrant IgM λ light chain accumulation, similar to Russell bodies. B‐cell lymphoma with Mott cell differentiation was diagnosed based on cytologic, histopathologic, and immunohistochemical features. This is the first report of B‐cell lymphoma with Mott cell differentiation in a cat.     

Survival and developmental competence of bovine embryos at different developmental stages and separated blastomeres after vitrification in different solutions

Generating techniques to enhance the success of blastomere separation is important for bovine economy, because it increases the number of transferable embryos. This study aimed to identify the optimum cryoprotectants for the vitrification of bovine embryos and the separation of blastomeres at different stages. In experiment 1, expanded blastocysts were vitrified in two different vitrification solutions, either (1) ethylene glycol (EG) + propylene glycol (PG) or (2) EG. The survival rate of blastocysts in the EG + PG was higher than that of the EG. In experiment 2, intact two‐cell and eight‐cell stage embryos were vitrified in the same solutions used in experiment 1. The EG + PG produced more dead embryos than the EG (P < 0.05). In the EG, the rate of blastocyst formation was similar for the vitrified two‐ and eight‐cell embryos and the non‐vitrified ywo‐cell embryos. In experiment 3, separated blastomeres of two‐ and eight‐cell embryos were vitrified in EG. There was no difference in the rate of blastocyst formation and total number of cells between the two vitrified groups. In summary, at the blastocyst stage, EG + PG was superior, based on both survival rates and cell numbers; however, at the 2–8 cell stage, the use of EG alone was better than the other groups.     

Penile Extramedullary Plasmacytoma in a Dog

Penile tumours are rare in dogs. Reported herein is a case of a penile extramedullary plasmacytoma (EMP) in a 5‐year‐old male cocker spaniel that was brought to a local hospital for an evaluation of a penile mass. The mass was approximately 1.3 cm in diameter at the time of presentation. In fine needle aspiration and histopathological examinations, the neoplastic cells showed eccentric round nuclei, a moderate amount of basophilic cytoplasm, and a peri‐nuclear clear zone, consistent with plasma cell morphology. There was nuclear pleomorphism with mononuclear giant cells and occasional bi‐nucleation. Round cells on the periphery of the mass demonstrated plasmacytic differentiation. Immunohistochemically, the neoplastic cells stained positive for MUM1 and light lambda chain. Based on the cytological and pathological observations, a diagnosis of penile EMP was established.     

Cellular Composition and Viability of Cloned Bovine Embryos Using Exogene-Transfected Somatic Cells

The present study compared the efficiency of transgenic (TG) cloned embryo production by somatic cell nuclear transfer (SCNT) with fetal-derived fibroblast cells (FFCs) which were transfected with pEGFP-N1 to in vitro-fertilized (IVF), parthenogenetic and SCNT counterparts by evaluating the rates of cleavage and blastocyst formation, apoptosis rate at different developmental stages, cell number, ploidy and gene expression in blastocysts. In SCNT and TG embryos, the rates of cleavage and blastocyst formation were significantly lower (p < 0.05) than those of IVF controls, but it did not differ between SCNT and TG embryos. In IVF control, 86.7% embryos displayed diploid chromosomal complements and the rates were significantly (p < 0.05) higher than those of SCNT and TG embryos. Most TG embryos (79%) with FFCs expressed the gene by both PCR and under fluorescence microscopy. The expression of apoptosis by TUNEL was first detected at six to eight cell stages in all embryos of IVF, SCNT and TG groups, but the expression rate at each developmental stages was significantly higher (p < 0.05) in SCNT and TG embryos than in IVF counterparts. The expression rate in inner cell mass (ICM) of TG embryos was significantly higher (p < 0.05) than in SCNT and IVF embryos. These results indicate that the high occurrence of apoptosis observed in SCNT and TG embryos compared with IVF counterparts might influence the developmental competence. Moreover, the SCNT embryos derived using non-transfected donor cells exhibited a lower apoptosis expression in ICM cells than in TG embryos derived using pEGP-N1-transfected donor cells suggesting a possible role of negative gene effect in TG embryos.     

Ultrastructural study of globidian parasites infecting the abomasum of sheep in Germany.

Globidian parasites infecting the abomasum of sheep in Germany were investigated by means of electron microscopy. The frequency of infection was found to be 93 %. The globidian cyst-like bodies contained multinucleate schizonts, developing merozoites or fully developed merozoites. Among the latter there were two different types, namely short and long forms. The process of merozoite formation was described in detail. The giant schizonts were subdivided into multinucleate cell portions of irregular size and shape. Their nuclei were then arranged at the periphery of the cell portions and underwent their last division which was combined with the differentiation of merozoites. The long form merozoites were elongated cylindrical in shape with terminal nucleus. They measured 7.7 μm in length and 1.0 μm in width. The merozoites of the short type were spindle-shaped with a central nucleus. They were 5.0 μm long and 1.0 μm wide. The globidian parasites were located in a parasitophorous vacuole of an intact host cell.     

Cryopreservation of bovine somatic cell nuclear-transferred blastocysts: effect of developmental stage

The effect of developmental stage on the survival of bovine somatic cell nuclear-transferred blastocysts after freezing and thawing was evaluated. We also investigated how freezing affects nuclear-transferred (NT) embryos and in vitro fertilized (IVF) bovine embryos. Advanced-stage bovine NT blastocysts survived freezing better than early-stage NT blastocysts (86 vs 14%). The trend was similar with IVF embryos (87 vs 30%). At the stages tested, there was no significant difference in the survivability of NT and IVF embryos from advanced (86 vs 87%) or early-stage blastocysts (14 vs 30%). The average survival rate did not differ between NT and IVF bovine embryos (50 vs 51%). The higher survival rate of advanced-stage blastocysts compared to early-stage blastocysts in NT and IVF bovine embryos might be due to their higher cell number. In NT (128 +/- 25 vs 53 +/- 20) and IVF (128 +/- 29 vs 75 +/- 22) groups, advanced-stage blastocysts contained a significantly higher total cell number than early-stage blastocysts. There was no difference in total cell number between advanced-stage NT and IVF blastocysts (128 +/- 25 vs 128 +/- 29), however, early-stage NT and IVF blastocysts (53 +/- 20 vs 75 +/- 22) differed significantly.