A morphological study of the thyroid gland in Risso's Dolphin,Grampus griseus

To accumulate histological information of cetaceans and basic information about metabolic systems of marine mammals, the thyroid gland of Risso's dolphins was examined by gross anatomical and light and electron microscopic observations. Gross anatomically, right and left lobes of the thyroid were not clearly discriminated, and no isthmus was observed. By light microscopy, irregular or oval follicular lumens were seen, and surrounded by follicular epithelial cells. By electron microscopy, the rough endoplasmic reticulum (rER) was seen adjacently to mitochondria at the basal and lateral regions of the follicular epithelial cells. RERs at the basal side of the cells sometimes contained flocculent material with the same electron density as the follicular lumen component. Microvilli were poorly developed at the apical surface of the cells. In the apical regions of the cells, there were typical Golgi complexes, multivesicular bodies, and granules with various size and electron density. The parafollicular cells were recognized among the follicular epithelial cells and in the interstitial regions but never protruded into the follicular lumen. These cells were present singly and/or formed clusters among the follicular epithelial cells, and often located adjacent to capillaries. They were obviously discriminated from follicular epithelial cells by higher electron density of their granules. In their cytoplasm, well-developed rERs, primary lysosomes, secondary lysosomes, multivesicular bodies, and phagosomes were recognized.     

Ultrastructural study of Langerhans cells in equine insect hypersensitivity "Kasen"

Ultrastructural features of Langerhans cells (LCs) of equine "Kasen" were studied. Electron microscopic observation revealed that LCs were dendritic and had irregular nuclear membranes. A number of Birbeck granules (Bgs) of various types were observed in the cytoplasm of LCs. In LCs in the upper stratum spinous, many Bgs were observed (Type 2 LC). LCs in the epidermo-dermal junction (EDJ) had a few Bgs, vesicles (multivesicular bodies) and highly electron-dense granular endosomes in the cytoplasm (Type 3 LC). Inactive LCs were also observed between the keratinocytes (Type 1 LC). Various types of LCs observed in the skin lesions of equine "Kasen" were interpreted as representing those that recognize, intake and process antigens.     

Production of giant mouse oocyte nucleoli and assessment of their protein content

Compared with advanced developmental stage embryos and somatic cells, fully grown mammalian oocytes contain specific nucleolus-like structures (NPB - nucleolus precursor bodies). It is commonly accepted that they serve as a store of material(s) from which typical nucleoli are gradually formed. Whilst nucleoli from somatic cells can be collected relatively easily for further biochemical analyses, a sufficient number of oocyte nucleoli is very difficult to obtain. We have found that isolated oocytes nucleoli fuse very efficiently when contact is established between them. Thus, well visible giant nucleoli can be obtained, relatively easily handled and then used for further biochemical analyses. With the use of colloidal gold staining, we estimated that a single fully grown mouse oocyte nucleolus contains approximately 1.6 ng of protein. We do believe that this approach will accelerate further research aiming at analyzing the composition of oocyte nucleoli in more detail.     

Changes in the morphology of nuclei in the oviduct epithelium of the cow during the estrous cycle

The changes in the structure of nuclei and nucleoli are good markers of the changes in the activity of epithelium cells of oviducts in cows during the sexual cycle. They are manifested by the changes in the proportion of condensed chromatin and interchromatin. In the secretory as well as ciliary cells, the maximum proportion of interchromatin is recorded in pro-oestrus and oestrus, whereas in metoestrus it is the proportion of condensed chromatin that increases. Metoestrus is characterized by the segregation of nucleoli and by the formation of ring-like nucleoli which partially persist also in dioestrus and in some cells throughout the cycle. Invaginations of the nuclear membrane (which are levelled off later) occur on many nuclei at the end of metoestrus and in dioestrus. Single nuclear bodies occur in the nuclei and the frequency of the the occurrence of interchromatin granules changes. Dilation of perinuclear spaces can be observed in the secretory cells, mainly in the period of oestrus.     

Immunohistochemical characterization of a hepatic neuroendocrine carcinoma in a cat.

A hepatic mass was identified in a 5-year-old, female mixed-breed cat that died spontaneously after a clinical history of progressive emaciation, ptyalism, and persistent coryza. At necropsy, a 7-cm-diameter, yellow-brown, firm, multilobulated tumor was identified in the liver. Microscopically, the mass consisted of neoplastic cells arranged in small, closely packed nests within a thin fibrovascular stroma. These cells were of medium sized and polygonal, with fine argyrophilic cytoplasmic granules. Nuclei were predominantly round with finely stippled chromatin and indistinct nucleoli. Mitotic figures were numerous. Immunohistochemically, most of the neoplastic cells were immunoreactive for chromogranin A, neuron-specific enolase (NSE), and cytokeratin AE1/AE3 and weakly labeled for synaptophysin. The tumor was negative for glial fibrillary acidic protein (GFAP), vimentin, and cytokeratins 5, 6, 8, and 17. Vascular emboli and intrahepatic micrometastasis were also identified with chromogranin A. All these features were consistent with a hepatic neuroendocrine carcinoma and emphasized the importance of using a panel of antibodies to diagnose such rare tumors.     

Uptake of ferritin by follicle-associated epithelium in the colon of calves.

Uptake of macromolecules (e.g., ferritin) by M cells in follicle-associated epithelium in small and large intestine was investigated in three healthy, conventionally raised, 2- to 3-week-old, female Holstein Frisian calves. A 2.5% solution of ferritin was injected into the ligated loops in mid-jejunum, in terminal ileum, in the ascending colon adjacent to the ileocecal junction, and in the proximal loop of the ascending colon containing gut-associated lymphoid tissue. After exposure times that ranged from 82 to 165 minutes, ferritin was detected in M cells of domes in the small intestine, as well as in cells in follicle-associated epithelium of proprial lymphoid nodules and lymphoglandular complexes of colon that morphologically resembled M cells of small intestine. Ferritin was found in apical invaginations, apical vesicles, multivesicular bodies, basal vesicles, and adjacent intercellular spaces. In addition to ferritin, apical vesicles, multivesicular bodies, and intercellular spaces contained 50-nm membrane-bound particles. More ferritin was endocytosed by M cells of the small intestine than by M cells of the large intestine. In the large intestine, higher amounts of ferritin were found in M cells of follicle-associated epithelium overlying proprial lymphoid nodules than in M cells of follicle-associated epithelium in the depth of lymphoglandular complexes. Based on these results, we concluded that M cells of follicle-associated epithelium in the colon of calves provide a route for antigen uptake into the intestinal lymphoid system.     

Ultrastructural morphology of lymphocytes of the Arabian oryx (Oryx leucoryx)

An ultrastructural study on the lymphocytes from peripheral blood samples from 20 healthy adult Arabian oryx (Oryx leucocoryx) was undertaken. Small lymphocytes ranged in size from 2–3.5 µm and exhibited the typical repertoire of organelles found in small lymphocytes of other animals but showed no evidence of azurophilic granules. Medium-sized lymphocytes were 5–6.5 µm in diameter and occasionally demonstrated azurophilic granules. Microvilli were a common finding of lymphocytes. Of particular interest was the presence of multivesicular bodies, which have previously only been described in human lymphocytes. Both small- and medium-sized lymphocytes of the Arabian oryx were smaller than those reported for other animals. Small lymphocytes exhibited short, thick microvilli, whereas medium-sized lymphocytes had long thin microvilli, a single nucleolus and occasionally azurophilic granules and multivesicular bodies.     

Electron microscopy studies on proliferation of foot-and-mouth disease virus in cell cultures. I. Morphologic changes in cell nucleus]

The first morphological indication of FMD infection of a cell culture was in the nucleus. Components of nucleoli became segregated and were finally present only as remnants. It was not possible to distinguish different stages of segregation, as in the case of entero-virus infections, because of the rapidity of FMD virus proliferation. Following changes in nucleoli there was margination of chromatin. Particularly striking was an increase in interchromatin granules. Changes in the nuclear membrane seemed to facilitate the transfer of nuclear material to the cytoplasm. Strongly pronounced dilatation of the peri-nuclear cleft, like that seen in aphthae and other tissues, were rarely visible in infected cell cultures.     

Spermiophagy in the spermatheca of Melipona bicolor Lepeletier, 1836 (Hymenoptera,Apidae, Meliponini)

The presence of spermatozoa in vesicles in the cytoplasm of the epithelial cells that constitute the spermathecal wall of Melipona bicolor queen (Meliponini) is discussed in relation to the organ structure. The epithelial wall is lined by an apparently continuous cuticle in the luminal surface that should be a non-transposable barrier to the luminal spermatozoa. However, some spermatozoa were seen crossing the cuticle through interruptions that was first interpreted as sectioning defects. Nevertheless, the sperm cells in well-structured cytoplasmic vesicles, bound by membranes and sometimes associated to multivesicular bodies, as well as cytoplasmic structures representative of intracellular digestion and the occurrence of the phenomenon in two of the three spermathecae studied, suggest a real spermiophagic hole in the spermathecal epithelial cells.     

Langerhans' cells in equine cutaneous papillomas and normal skin.

Langerhans' cells (LC) were investigated immunohistochemically and electron microscopically in normal equine epidermis and 133 equine cutaneous papillomas experimentally induced in five 2-year-old Thoroughbred horses. Class II major histocompatibility complex antigen-positive dendritic LC were found in the normal epidermis and ultrastructurally had the characteristic Birbeck's granules. In the developing phase of the papillomas, LC were significantly decreased in number and size, indicative of a hypofunctional state. In the regressing phase of the papillomas, LC were markedly increased in number, especially at the epidermis-dermis junction. LC with long dendrites were rich in cytoplasm with well-developed cytoplasmic organelles, including Golgi apparatus, lysosomes, Birbeck's granules, and multivesicular bodies. These LC were hyperfunctional. An infiltration of many T lymphocytes was also observed at the epidermis-dermis junction.     

Structure of the lining epithelium of the cauda epididymis of the golden hamster

The ductus epididymis has roles in the maturation and storage of spermatozoa. The main function of the cauda epididymis is the storage of spermatozoa; however, this region exerts other morphophysiological roles. So, this study was aimed at investigating structural features of the cauda epididymis epithelium, which could indicate roles other than the storage. The relative percentages of the cell types in the epithelium were 74.9, 6.9, 12.5 and 5.6% of principal, clear, basal and halo cells respectively. Large intercellular spaces were seen among the lateral plasmatic membranes of adjacent principal cells or among these cells and others cell types. These spaces were found to be filled with multivesicular bodies, myelin figures, scrolls and debris of membranes or flocculent dense material. Clear cells had the cytoplasms filled with lysosomes (¾ of basal cytoplasm), and vacuoles and vesicles (¼ of apical cytoplasm). The observations allowed us to infer that clear cells could act in the process of endocytosis and also in water transfer from the lumen to the interstitium through the epithelium compartment. Moreover, transcytosis may occur at the cauda epididymis of Golden hamster.     

Ultrastructural observations on the platelets of the Arabian oryx (Oryx leucoryx)

An ultrastructural study on the platelets from peripheral blood samples from 20 healthy adult Arabian oryx (Oryx leucocoryx) was performed. Characteristic was the extreme polymorphism of both the platelets, as well as their alpha granules. They vary in size from 100 to 800 nm in diameter and their numbers typically are less than those reported for humans and other animal species. Also, the alpha granules in contrast to those of humans and animals, such as the Arabian tahr, do not have nucleoids. Typically, the oryx platelets exhibit only 1–2 electron‐dense bodies per cell and they lack an open canalicular system. Of particular interest is the presence of Type I multivesicular bodies, which have previously only been described in human megakaryocytes and are hypothesized as being intermediate development stages of alpha and dense granules.     

Ultrastructural Aspects of the Prenatal Bovine Ovary Differentiation with a Special Focus on the Interstitial Cells

The aim of this investigation was to study the ultrastructural features during the development of fetal bovine ovaries (crown rump length ranging from 11.4 to 94.0 cm). An interesting observation was the occurrence of big elongated cells containing a variety of electron dense granules and light homogenous vacuoles/bodies. They were located between the stroma cells surrounding the germ cell cord ends, adjacent to the first formed primordial follicles, typically situated near blood vessels. ER alpha and ER beta receptor positive cells could be detected in the same regions by means of immunohistochemistry. Intercellular bridges linked the germ cells nests oogonia. Germ cell cords consisted of centrally located, large, pale oogonia, surrounded by elongated somatic cells with very long cytoplasm extensions. Primordial follicles with flat pale follicular cells could be observed on the inner end of the cords. Extrusions of the outer nuclear membrane could often been recognised in voluminous oocytes.     

Sub-cellular localisation of alkaline phosphatase activity in the cytoplasm of tammar wallaby (Macropus eugenii) neutrophils and eosinophils

Alkaline phosphatase (ALP) has been used in studies of neutrophil morphology and function as a marker for identifying different granule populations. In human neutrophils, ALP is found within secretory vesicles, a rapidly mobilisable vesicle population important for upregulating membrane receptors during early activation. Intra-cellular ALP activity in the heterophils of rabbits and guinea pigs, in contrast, is found only in secondary granules. The neutrophils and eosinophils of tammar wallabies (Macropus eugenii) have previously been reported to contain large amounts of ALP activity when stained using routine cytochemical techniques. To define the subcellular location of ALP in this species, cell suspensions were examined using cerium chloride cytochemistry and transmission electron microscopy (TEM). ALP was found in 2 distinct cytoplasmic compartments. One compartment displayed morphology consistent with a subpopulation of secondary granules while a second tubulo-vesicular population appeared similar to the secretory vesicles of human neutrophils. Thin tubular vesicles containing ALP were also identified within the cytoplasm of tammar wallaby eosinophils. Large numbers of ALP-containing vesicles have not been recognised previously in eosinophils and this may represent a novel cytoplasmic compartment. In both cell types, ALP-containing structures showed alteration in morphology following stimulation with N-formyl-Met-Leu-Phe (fMLP) and PMA.     

Intracranial meningioma with polygonal granular cell appearance in a Chihuahua

A menigioma with polygonal granular cell proliferation in an 11-year and 8-month-old male Chihuahua is described. The tumor was observed under the dura matter of the right cerebrum. Microscopically, the tumor consisted of solid growth foci of small- or large- sized polygonal cells, with pale-stained nuclei, prominent nucleoli, and fine granular to foamy eosinophilic cytoplasm. Some of the proliferating cells contained variable amounts of cytoplasmic PAS-positive granules. Immunohistochemical analysis revealed that neoplastic cells were positive for vimentin and S-100 protein. Ultrastructurally, the neoplastic cells contained vesicular structures with a few small round-shaped bodies in the cytoplasm. We diagnosed the case as canine meningioma with granular cell appearance.     

A subependymal giant cell astrocytoma in a cat

A 6-year-old spayed female Domestic Shorthair cat presented with a 1 to 2-month history of blindness and altered behavior. At necropsy, a 1-cm-diameter, firm white mass was found arising from the subependymal region of the right lateral ventricular wall that protruded into and partially filled the lumen. Histologically, there was a well-demarcated, expansile paraventricular neoplasm composed of moderately pleomorphic cells within a richly fibrillar matrix arranged in interlacing streams and perivascular pseudorosette-like patterns. Neoplastic cells varied in morphology from small spindloid cells to larger polygonal cells with eccentric vesicular nuclei to neuronlike cells with vesicular nuclei and prominent nucleoli. The mitotic index was low. Immunohistochemically, neoplastic cells were positive for S-100 protein, glial fibrillary acidic protein, and neuron-specific enolase and negative for neurofilament protein. Ultrastructurally, the cells contained few to abundant bundles of intermediate filaments with variable numbers of mitochondria, endoplasmic reticulum, and ribosomes. These features are characteristic of subependymal giant cell astrocytoma (SEGA) in humans. To our knowledge, this is the first reported case of SEGA in domestic animals.     

Ultrastructural morphology of leukemic cells from 14 dogs

Transmission and/or scanning electron microscopic features of leukemic cells from seven dogs with myelomonocytic leukemia, four dogs with monocytic leukemia, and three dogs with lymphocytic leukemia were studied. Few ultrastructural features distinguished normal cells from leukemic cells. Nuclear to cytoplasmic asynchrony, abnormal cytoplasmic granules, nuclear pockets, and cytoplasmic fibrillar bundles were seen more frequently in leukemic cells. Small ridges or ruffles and a few microvilli were surface characteristics of cells from dogs with myelomonocytic leukemia. An occasional cell had large undulating ruffles. Cells from dogs with monocytic leukemia and lymphocytic leukemia usually had smoother surfaces.     

Electron microscopic studies of estrogen-induced ciliogenesis and secretion in uterine tube of the gilt.

The time required for occurrence of estrogen-induced uterine tubal (oviductal) ciliogenesis and for differentiation of secretory cells was studied, utilizing electron microscopy procedures. Sixteen cycling gilts were ovariectomized; 3 to 4 months later, 12 principal gilts were each given subcutaneous injections of 17 beta-estradiol in 0.5 ml of corn oil at the rate of 200 mug/day, and 4 control gilts were given injections of corn oil only at the rate of 0.5 ml/day. Two principals each were killed on days 1, 2, 3, 4, 5, and 7 after start of treatment. The epithelial heights were low and completely atrophied 3 to 4 months after ovariectomy. Uterine tubal cilia were absent in all the control gilts. Cytologic changes were not seen in the atrophied epithelium of ovariectomized gilts 1 day after estradiol treatment, but definite proliferative elements consisting of an extensive fibrillar meshwork encrusted with granules (60 to 80 nm) were observed in close association with the nuclear envelope and in the apical cytoplasm after 2 days of estradiol treatment. By day 3, enlarged electron-opaque granules referred to as condensation forms, undergoing various stages of depletion, were closely associated with radially arranged procentrioles. These associations have been referred to as generative complexes. The presence of many generative complexes indicates that maximal production of basal bodies can be expected after 3 days of treatment with estradiol. The depletion of the condensation forms produced hollow spheres with thin walls as the procentrioles grew in length and assembled their microtubules. Enlarged mature-appearing basal bodies were abundant in the cytoplasm after 3 days of estradiol treatment. These bodies aligned themselves linearly along the luminal surface of the cell. Small ciliary buds were then formed above the cell surface, and ciliary filamentogenesis occurred in the bud. Motile cilia were observed on day 3, but cilia numbers increased markedly between day 4 and days 5 and 7. Procentrioles were generated from the diplosomal centriole after 2 days of estradiol treatment. These observations have provided evidence for both ancentriolar and centriolar basal body replication in the ciliated cells of uterine tube of the gilt. Maximal secretory cell differentiation occurred after 3 days of estradiol treatment. Hypertrophy of cytoplasmic organelles was evident on day 3, but the number of secretory granules and amount of rough endoplasmic reticulum increased markedly on days 5 and 7. Close association of secretory granules, Golgi apparatus, and endoplasmic reticulum was evident after estadiol treatment. These data indicate that both ciliated and secretory cells are sensitive to estrogen.     

Bovine ileal dome lymphoepithelial cells: endocytosis and transport of Brucella abortus strain 19

Ligated ileal loops of calves were inoculated with Brucella abortus and examined at 2, 4, 6, 10, and 24 hours post-inoculation. B. abortus was identified by light and electron microscopy using immunoperoxidase and antibody-coated colloidal gold techniques. B. abortus was detected in vesicles, phagolysosomes, and large vacuoles of lymphoepithelial cells. Numbers of intracellular bacteria decreased with time after inoculation. B. abortus was also seen between and below lymphoepithelial cells and free in the dome interstitium and intestinal lymph vessels. Neutrophils and macrophages in both epithelium and lamina propria contained intact or degraded bacteria within phagosomes, phagolysosomes, and multivesicular bodies. These studies showed that (1) transepithelial migration of B. abortus occurred principally by dome lymphoepithelial cell endocytosis and transport, and (2) B. abortus was degraded by macrophages and neutrophils of the gut-associated lymphoid tissue.