A histochemical study of amines in palatal tissue from normal and glycosuric mice

Synopsis A fluorescence histochemical technique for localizing catecholamines was applied to palatal tissue obtained from laboratory mice and from glycosuric and nonglycosuric Australian hopping mice. Specific catecholamine fluorescence related to blood vessels was demonstrated in all the tissue samples, though there was a tendency for this to be less marked in palates taken from the glycosuric animals.Although the presence of discrete areas of bright yellow fluorescence was noted in the tissue of the laboratory mice, these areas of yellow fluorescence (probably serotonin within mast cells) were virtually absent from the palates of the hopping mice despite the fact that there was little difference in the respective mast cell populations in the palatal tissues of the laboratory and hopping mice as determined by metachromatic staining procedures.     

A morphologic and histochemical study of the mesentery in the guinea pig

The guinea pig mesentery is a uniform, continuous, thin (18 micron) sheet of connective tissue covered by a single layer of flattened mesothelial cells on both surfaces. Tight and gap junctions provide for cell-to-cell adhesion among mesothelial cells. These cells possess numerous micropinocytotic vesicles; a conspicuous basal lamina separates the mesothelium from the underlying connective tissue. Most of the material found between the two serous coverings consisted of a three-dimensional meshwork of abundant collagenous fibers intermingled with a sparse net of very thin (0.4 micron) elastic fibers. Two distinct populations of collagen fibrils are segregated into different compartments of the mesentery. One population is formed of thick (56 nm) fibrils which associate to form closely packed fibers. The second population, composed of loosely arranged thin (38 nm) fibrils which do not become assembled into fibers, is found underlying the basal lamina that separates the mesothelium from the connective tissue. These observations strongly suggest that the mesentery contains both collagens type I and type III. The guinea pig mesentery contains 6.8 mg of sulfated glycosaminoglycans/g dry weight. Most of these glycosaminoglycans (78%) were identified as dermatan sulfate, whilst the rest (22%) corresponded to heparan sulfate.     

A histochemical study of liver enzymes involved in glycogen metabolism in the X-irradiated rat

Summary The liver enzymes responsible for the breakdown and synthesis of glycogen from glucose have been investigated cytochemically in rats exposed to 1200 rads of x-irradiation. It was found that significant changes occur in their activities and that amylophosphorylase and amylo-1,6-glucosidase (debranching enzyme), both of which are responsible for the conversion of glycogen to glucose, are markedly inhibited by radiation. A significant inhibition of the activity of 1,41,6 transglucosidase (branching enzyme) was also observed. In contrast, the activity of UDPG-glycogen transglucosylase, which is responsible for the in vivo synthesis of 1,4-polysaccharides, was found to be stimulated.     

Molecular and morphologic changes during the epithelial-mesenchymal transformation of palatal shelf medial edge epithelium in vitro.

The fate of the medial edge epithelial (MEE) cells during palatal fusion has been proposed to be either programmed cell death or epithelial-mesenchymal transformation. Vital cell labeling techniques were used to mark the MEE and observe their fate during palatal fusion in vitro. Fetal mouse palatal shelves were labeled with Dil and allowed to proceed through fusion while maintained in an organ culture system. The tissues were examined at several stages of palatal fusion for the distribution of Dil, presence of specific antigens and ultrastructural appearance of the cells. The MEE labeled with Dil occupied a midline position at all stages of palatal fusion. Initially the cells had keratin intermediate filaments and were separated from the underlying mesenchyme by an intact basement membrane. During the process of fusion the basement membrane was degraded and the Dil-labeled MEE were in contact with the mesenchymal-derived extracellular matrix. In the late stages of fusion the Dil-labeled MEE altered their cellular morphology, had vimentin intermediate filaments, and were not associated with an identifiable basement membrane. Dil-labeled cells, without an epithelial phenotype, remained present in the midline of the completely fused palate. The data indicate that the MEE did not die but underwent a phenotypic transformation to viable mesenchymal cell types, which were retained in the palatal mesenchyme.     

Synaptonemal complex alterations in X-irradiated and in oestrogen-treated mice: a comparative study.

Synaptonemal complexes (SCs) were analysed in male NMRI mice either X-irradiated or treated with oestradiol benzoate (E2B). Animals 30 days old underwent a single X-ray exposure of either 5, 7.5 or 10 Gy and were killed at different times after exposure, i.e., 24 h, 1, 4, 12 and 16 weeks. E2B was injected daily to adult mice from day 30 to day 60 or up to day 90 of age. Oestradiol was also administered during the neonatal period and animals were examined on days 28, 60 and 90 of age. Different SC alterations were found in X-irradiated and in E2B-treated mice. SC lesions were rare in oestrogen-treated adult mice. Among SC anomalies, asynapsis and fragmentation of SC were common lesions. However, the former was more frequent in E2B-treated mice, whereas the latter was more frequent in X-irradiated mice. Quadri- or multi-valents, bridges between bivalents, rings and loops were exclusively encountered in the latter, whereas heterotelomeric associations seemed to be specific in E2B-treated animals. The mechanisms of the different SC lesions are discussed.     

Fine structural changes and localization of phosphatases in the epithelium of the duodenal crypt of X-irradiated mice

Summary The ultrastructural localization of acid phosphatase has been studied in the stem cells of duodenal crypt of X-irradiated mice. After a transitional increase of the cytochemically detectable activity of this enzyme, in the first hours after irradiation, the reaction appears less and less important in the cells. These observations are in agreement with the morphological alterations of the ultrastructures provoked by the irradiation. The meaning of these modifications is discussed.This work was done thanks to the contract C.E.N./A.I.E.A. No. 347/RB and thanks to grants from the Fonds de la Recherche Scientifique Fondamentale Collective.     

Tracheobronchial epithelium in the adult rhesus monkey: a quantitative histochemical and ultrastructural study

Previous studies of the intrapulmonary conducting airways of sheep and rabbit have demonstrated marked diversity in the epithelial populations lining them. Because studies of trachea and centriacinar regions of macaque monkeys suggested that primates may be even more diverse, the present study was designed to characterize the epithelial population throughout the airway tree of one primate species, the rhesus monkey. Trachea and intrapulmonary airways of the right cranial and middle lobes of glutaraldehyde/paraformaldehyde-infused lungs of five adult rhesus monkeys were microdissected following the axial pathway. Each branch was assigned a binary number indicating its specific location within the tree. The trachea and six generations of intrapulmonary airway from the right cranial lobe were evaluated for ultrastructure and quantitative histology as were those of the right middle lobe for quantitative carbohydrate histochemistry. Four cell types were identified throughout the tree: ciliated, mucous goblet, small mucous granule, and basal. The tallest epithelium lined the trachea; the shortest, the respiratory bronchiole. The most cells per unit length of basement membrane were in proximal intrapulmonary bronchi; the least, in the respiratory bronchiole. The nonciliated bronchiolar epithelial or Clara cell was restricted to respiratory bronchioles. Sulfomucins were present in the vast majority of surface goblet cells in the trachea and proximal bronchi. In proximal bronchi, neutral glycoconjugates predominated in glands and acidic glycoconjugates in surface epithelium. In terminal and respiratory bronchioles the ratio of acidic glycoconjugate to neutral glycoconjugate equaled that in proximal bronchi, although glands were not present. Sulfomucins were minimal in terminal airways. We conclude that the characteristics of the epithelial lining of the mammalian tracheobronchial airway tree are very species-specific. The lining of the rhesus monkey does not have the diversity in cell types in different airway generations observed in sheep and rabbit. Also, the populations lining these airways in the rhesus are very different from either the sheep or rabbit in number, proportions of different cell types, glycoconjugate content, and distribution of specific cell types.     

Effects of vitamin A-deprivation on hamster tracheal epithelium. A quantitative morphologic study

The effects of vitamin A-deprivation on the tracheal epithelium were studied in 35-day old hamsters that had been raised since birth on a vitamin A-deficient diet. Colchicine and 3HTdR were given 6 hours before death and the proliferative activities of basal cells and mucous cells were quantified separately by 3HTdR labeling indices and mitotic rates. Vitamin A-deprivation decreased replication of basal cells and mucous cells in tracheal epithelium which showed minimal morphologic change. The mitotic rates and labeling indices were reduced 3 to 4-fold in basal cells and 14-fold in mucous cells (analyzed as percent of total number of each cell type) compared with controls. Thus, replication of mucous cells was more inhibited by lack of vitamin A, than replication of basal cells. The disparate hypoplasia of basal cells and mucous cells in epithelium showing minimal change, resulted in a relative increase in the proportion of basal cells and a relative decrease in the proportion of mucous cells, which could be erroneously interpreted as "basal cell hyperplasia". Proportions of preciliated and ciliated cells were also decreased compared to controls. At foci of stratification and epidermoid metaplasia, cell replication rates were increased over controls and more than 70% of all mitotic activity was associated with "non-basal" cells. Genesis of these lesions was coincident with cell death and cell loss. The histogenesis of stratification and epidermoid metaplasia was characterized. Morphological evidence indicated that these lesions were closely related histogenetically and were composed, for the most part, of altered mucous cells which expressed dual phenotypes i.e. keratinization and mucus synthesis.     

A histochemical study of acid phosphatase in normal and virus-transformed cultured fibroblasts.

The distribution of acid phosphatase has been investigated in normal and virus-transformed cultured hamster and mouse fibroblasts. The enzyme was found to be present in lysosomes, autophagic vacuoles and elements of the Golgi apparatus. It was also found to be associated with a surface coat in some virus-transformed mouse cells and in the cytoplasm of both normal and transformed hamster cells.     

Restoration of mucociliary tracheal epithelium following deprivation of vitamin A. A quantitative morphologic study

In order to learn more about the respective roles played by basal cells and mucous cells in the maintenance of tracheal mucociliary epithelium, cell kinetics and epithelial cell morphology were characterized over a 7-day period, during which dietary vitamin A was restored to previously deprived hamsters. Hamsters were reared from birth to 35 days of age on vitamin A-replete or deficient diets. Deprived hamsters were made replete by 5 mg vitamin A-acetate orally, plus a vitamin A-replete diet. Colchicine and 3HTdR were given 6 h before death. The numbers of basal cells, mucous cells, preciliated cells and ciliated cells, and mitotic rates (MR) and labeling indices (LI) of basal cells and mucous cells, were quantified in glycol methacrylate sections stained with PAS-lead hematoxylin. Vitamin A-deprivation decreased replication of basal cells and mucous cells in tracheal epithelium which showed minimal morphological change. The proportion of basal cells was increased and proportions of mucous, preciliated and ciliated cells were decreased. Following restoration of vitamin A to the diet, the basal cell MR remained below control level throughout the experimental period, but the mucous cell MR started to rise on day 2-replete, and on day 3-replete and thereafter the mucous cell MR was within the control range. Basal cell and mucous cell LI's showed similar trends. Preciliated cells were reduced or absent in vitamin A-deprived epithelium. Their number had risen by day 3-replete and thereafter they were generated within the control range. These cells matured into ciliated cells. By day 4-replete, the proportion of basal cells had decreased markedly and the proportions of mucous cells, and preciliated plus ciliated cells had increased, so that at this time cellular proportions were within or near control values. This trend continued so that by day 7-replete, a nearly normal mucociliary epithelium was restored. The results show that vitamin A-levels modulate replication rates of basal cells and mucous cells and indicate that mitotic division of mucous cells is a prerequisite for the genesis of preciliated cells and new mucous cells and for restoration of the mucociliary epithelium following deprivation of vitamin A in the diet.     

Palatal shelf reorientation in hamster embryos following treatment with 5-fluorouracil

A study was undertaken to examine the issue of whether achieving a critical mass of cells and/or palatal shelf volume during vertical development of shelf is essential for reorientation to occur. In control and 5-fluorouracil (5FU)-treated hamster embryos' palatal shelves, at different times during gestation, the numbers of both epithelial and mesenchymal cells were counted and cross-sectional area was measured. DNA synthesis was measured by 3H-thymidine incorporation and was used as an index of growth by cell proliferation. The control data indicated that, unlike development during initial 24 hours, the later period of vertical palatal development was characterized by a steady level of mesenchymal and epithelial cell numbers and palatal shelf area. Following 5FU treatment all the measurements were reduced, and until they reached the equivalent of control values, the palatal shelves did not reorient. The density of mesenchymal cells in the developing palate did not seem to affect cell number. On the basis of the analysis of results of the present study, along with those reported in the literature, it is suggested that, in hamsters, acquisition and maintenance of both a specified number of mesenchymal cells and shelf area, at least 24 hours prior to reorientation, may be critical for ensuing mesenchymal differentiation to enforce palatal shelf reorientation on schedule. 5FU affected these features to delay reorientation of the palatal shelf.     

Fluorescence study of thymus lymphocytes of X-irradiated mice and their progeny.

The mechanisms of changes in the ultra-violet fluorescence (U.V.F.) intensity of mouse thymus lymphocytes 24 hours and 30 days after whole-body X-irradiation have been studied. The thymus lymphocytes of the first generation offspring (F1) from X-irradiated males and unirradiated females were also investigated. At 24 hours after irradiation the U.V.F. intensity decreased for small doses (50 and 65 rad) and increased for doses of more than 100 rad. The changes in U.V.F. intensity were related to a size-independent mechanism. It was found that the U.V.F. increase for doses of 100-700 rad was not connected with the appearance of non-viable (eosin test) cells. The changes in U.V.F. intensity and cellular composition of the thymus were the same 30 days after irradiation and for F1 mice. The increase in U.V.F. intensity was about 14% and did not depend on dose between 50 and 500 rad. About one-half of this increase was connected with an increase in the proportion of medium and large lymphocytes in the thymus. The rest of the effect was related to a size-independent mechanism.