Demonstration of the heterogeneity of the mast cell population on the basis of the mucopolysaccharide content.

From the aspect of its mucopolysaccharide content the mast cell population is not homogeneous. The pulmonary and heart muscle mast cells of the rat are alcian blue positive, the mast cells of the thyroid gland, lymph nodes, subcutaneous connective tissue, mesentery and peripheral nerve are safranin positive, whereas among the mast cells of the peritoneal cavity and the thymus there are both alcian blue and saffranin positive forms. The least acid mucopolysaccharides are in the mast cells of the peritoneal fluid, the mesentery and the lungs, whereas the most acid ones are in the mast cells of the lymph nodes, the subcutaneous connective tissue and the thyroid gland. There is a considerable difference between the two last mentioned organs. The mast cells of the subcutaneous connective tissue are end-product cells without amine or precursor turnover, whereas the mast cells of the thyroid gland incorporate and deliver amines, which may participate in the regulation of the host gland.     

Cell Migration and Induction in the Development of the Surface Ectodermal Pattern of the Xenopus laevis Tadpole

The surface of the Xenopus tadpole contains three specialized, transient cell types; the ciliated, hatching gland, and cement gland cells. To distinguish whether the appearance of these cell types on the surface is due to induction of surface cells or due to migration of deep ectodermal cells into the surface, we transplanted labelled surface or deep cells to unlabelled hosts at early to mid-gastrulae. After raising the host to a tadpole (Stage 28), we examined the embryo's surface for ciliated, hatching gland, and cement gland cells, and observed which cells were labelled. We find that all ciliated cells move into the surface from the deep ectodermal layer along with other cells of unknown function. Hatching gland cells arise by induction of surface cells as do the majority of cement gland cells. A few deep cells give rise to cement gland cells. Therefore, migration of deep cells to the surface and localized induction of surface cells contribute to the final surface patterning of the Xenopus tadpole.     

Cytophotometric analysis of the cell population composition of the the preoptic area of the hypothalamus in the common frog

DNA contents in squashed cells of the adult frog hypothalamic preoptic region (HPR) were measured using the Feulgen and UV cytophotometry techniques. The histone-DNA ratio in the cell nucleus was determined by means of a combined Feulgen-heparin-Alcian blue staining procedure. The nuclei of the vast majority of HPR cells have a diploid DNA content. However, in cells of this group the mean values of DNA amount and the distribution range were always higher than those in hepatocytes used as a diploid standard. Such a heterogeneity in DNA content in the diploid part of HPR cell population could apparently suggest some differences in the nuclear chromatin arrangement to be always higher in spring before the frog spawning, and it seems to be characteristic of this type of cells. About 1 per cent of cells with hyperdiploid surplus of DNA (H2c cells) as well as of tetraploid cells (4c and 2c X 2 cells) is found in HPR in frogs sacrificed both in winter and in summer. The quota of these cells has no reference either to the frog's age or to the annual cycle. The fact that the mean DNA values in H2c and 4c cells are much higher than in the standard cannot be explained by the presence of different amounts of nuclear proteins only. It is suggested that at least some part of the highest DNA values may be due to an actual extra DNA synthesis in a small constantly existing pool of HPR cell population.     

Motile cells in the mucosa of the cloacal urodaeum and proctodaeum of the hen

Summary Motile cells (mast cells, granulocytes, lymphoid cells) are described in the mucosa of the cloacal urodaeum and proctodaeum of the female domestic fowl. Diffuse lymphoid tissue with lymphatic nodules occurs in the urodaeum at the ureteral ostium. Small local aggregations of lymphoid tissue can be observed in the mucosa of the proctodaeum. Cells originating from these sites penetrate the basal lamina of the epithelium and are then found between the epithelial cells.In the subepithelial layers the motile cells sometimes are in contact with each other. Mast cells (tissue basophils) form contact zones, resembling desmosomes or half desmosomes, with smooth muscle cells. In the mast cells three types of granules can be distinguished. Their ultrastructure is discussed in comparison with that in similar cells of the guinea pig.Dedicated to Professor Dr. med. M. Watzka in honor of his 75th birthday     

Immunohistochemical localization of the transferrin receptor in the seminiferous epithelium of the rat

The transferrin receptor has been immunohistochemically localized in the seminiferous epithelium of the rat with a monoclonal antibody, MRC OX26, which recognizes the transferrin receptor glycoprotein. The receptor was detectable on mitotically and meiotically dividing germ cells and, less abundantly, on round spermatids. It was lost from germ cells during spermatid elongation and was undetectable on immature spermatozoa. The transferrin receptor was also present on Sertoli cells in the testes of immature animals and on Sertoli cells in the testes of aspermatogenic animals that had been irradiated in utero. It was not detectable on Sertoli cells in the testes of cryptorchid animals. These studies demonstrate that the transferrin receptor is abundant on dividing germ cells as well as dividing somatic cells.     

Development of the mammalian gonad: the fate of the supporting cell lineage

Sex determination in mammals is mediated via the supporting cell lineage in the fetal gonad. In the very early stages of gonadal development, the fate of the supporting cell population is critically dependent on the expression of the male-determining gene on the Y chromosome. If this gene is absent or fails to be expressed, or is expressed too late or in too small a number of supporting cells, all supporting cells (XX or XY) differentiate as pre-follicle cells and development proceeds along the female pathway. Supporting cells in which the male-determining gene is expressed in a timely manner differentiate as pre-Sertoli cells; given sufficient such cells, testis cords form and development proceeds in a male direction. If XX supporting cells are also present, a few may be recruited into the pre-Sertoli population and participate in testis cord formation. The subsequent fate of pre-follicle cells depends critically on interaction with the germ cell population in the developing gonad: absence of germ cells may lead to partial masculinization of the gonad, and/or to disappearance of the supporting cell component.     

Ultrastructure of the cardiac and pyloric glands of the gastric mucosa of the South African hedgehog,Atelerix frontalis

The cardiac and pyloric glands in the gastric mucosa of the South African hedgehog, Atelerix frontalis, are described. The cardiac area of the stomach contains proper cardiac glands and lacks undifferentiated fundic glands. The cardiac glands are simple tubular, coiled, and lined with columnar cells ultrastructurally similar to those of the gastric surface epithelium. Secretory granules with varying electron densities fill the apical cytoplasm of these cells. In contrast to other mammals, these glands lack mucous neck cells. The neck of the pyloric glands contains only a single cell type, whereas the basal regions of these glands contain “light” and “dark” cells. The secretory granules in the “dark” cells and the pyloric neck cells have a moderate electron density and often contain an electron dense core. An electron-lucent cytoplasm with numerous polysomes is characteristic of the “light” cells. Some “light” cells contain electron-dense granules in the apical cytoplasm. The presence of only neutral mucins in the cardiac gland cells denotes the absence of mucous neck cells. The acidic mucins within the pyloric neck cells seem to indicate that these cells are mucous neck cells, whereas the neutral mucins within the basally located pyloric gland cells show at least a partial functional difference from the pyloric neck cells. © 1993 Wiley-Liss, Inc.     

Culturing the epiblast cells of the pig blastocyst

Summary Pig epiblast cells that had been separated from other early embryonic cells were cultured in vitro. A three-step dissection protocol was used to isolate the epiblast from trophectoderm and primitive endoderm before culturing. Blastocysts collected at 7 to 8 days postestrus were immunodissected to obtain the inner cell mass (ICM) and destroy trophectodermal cells. The ICM was cultured for 2 to 3 days on STO feeder cells. The epiblast was then physically dissected free of associated primitive endoderm. Epiblast-derived cells, grown on STO feeders, produced colonies of small cells resembling mouse embryonic stem cells. This primary cell morphology changed as the colonies grew and evolved into three distinct colony types (endodermlike, neural rosette, or complex). Cell cultures derived from these three colony types spontaneously differentiated into numerous specialized cell types in STO co-culture. These included fibroblasts, endodermlike cells, neuronlike cells, pigmented cells, adipogenic cells, contracting muscle cells, dome-forming epithelium, ciliated epithelium, tubule-forming epithelium, and a round amoeboid cell type resembling a plasmacyte after Wright staining. The neuronlike cells, contracting muscle cells, and tubule-forming epithelium had normal karyotypes and displayed finite or undefined life spans upon long-term STO co-culture. The dome-forming epithelium had an indefinite life span in STO co-culture and also retained a normal karyotype. These results demonstrate the in vitro pluripotency of pig epiblast cells and indicate the epiblast can be a source for deriving various specialized cell cultures or cell lines.     

An ultrastructural study of the regenerating breast feather of the fowl

The developmental morphology of regenerating male breast feathers of the jungle fowl was studied at the ultrastructural level. The process of keratinization was observed in the three types of cells which form feather barbs: barbule cells, cortical cells, and medulla cells. Keratinization first became evident in the barbule cells and resembled the process of keratinization as observed in hair cortical cells and embryonic down feathers. Eventually the whole cytoplasmic area of the barbule cell was occupied by keratin. The barb cortex cells became keratinized in a similar fashion as the barbule cells but not until they were developmentally twice as old as the barbule cells. When keratinization was complete in these cells, the keratin was in the form of large agglomerates scattered in the cytoplasm. The barb medulla cells showed no obvious signs of keratinization until they were developmentally three times as old as the barbule cells. Keratin filament bundles were first seen near the plasma membranes of the medulla cells. Large empty vacuoles appeared in the cytoplasm which also contained moderate amounts of glycogen.     

Ultrastructure of the Testicular Interstitial Tissue of the Hagfish Eptatretus stouti

Ultrastructure of the somatic components of the testis of the Pacific hagfish was studied. Interstitial cells, equivalent to Leydig cells of higher vertebrates, containing smooth-surfaced endoplasmic reticulum and mitochondria with tubular cristae were found in the interstitial tissue, as well as leucocytes, fibroblasts and other cells of unknown role. Two kinds of somatic cells were observed in the testicular follicles: Sertoli cells and “stellate cells”. The significance of interstitial cells was discussed in relation to possible involvement in steroidogenesis.     

Loss of polar trophoblast during differentiation of the blastocyst of the horse

Twelve blastocysts, collected 7-12 days after ovulation (Day 0), were examined by light and electron microscopy to investigate the nature of the relationship of the polar trophoblast (Rauber's layer) to the inner cell mass. On Day 7, the polar trophoblast was intact and formed a flattened layer overlying the epiblast cells of the inner cell mass. As blastocysts enlarged to greater than 1 mm in diameter, small discontinuities appeared in the polar trophoblast, where epiblast cells intruded onto the surface. At this time, trophoblast cells adhered closely to adjacent and underlying epiblast cells, forming an irregular layer of cells capping the epiblast. With continued increase in blastocyst size, polar trophoblast cells became isolated but maintained their characteristic apical endocytic structures. By Days 10-12, the scattered trophoblast cells showed evidence of deterioration, and vacuoles containing cell debris were common within the epiblast. It is suggested that polar trophoblast cells become scattered, rather than withdrawing as a unit, because they become more adherent to subjacent epiblast cells than to adjacent trophoblast cells. It is further suggested that most of the isolated cells are eventually phagocytosed by epiblast cells.     

Morphological characterization of stationary reticulum cells of the stromal in the mesenteric lymph node of the guinea pig

The large mesenteric lymph node taken from guinea pigs in a period of time ranging from the 10th day prepartum till the 26th day postpartum has been examined in order to study: the morphological features of the stromal stationary reticulum cells with particular regard to recognize their stages of development; the possible ontogenetic relationship between these cells during the maturation of the lymphoid tissue. Our data support the hypothesis that from local mesenchymal cells originates a pool of poorly differentiated reticulum cells that can give rise to stromal stationary reticulum cells (myofibroblast-like cells, fibroblast-like cells, pericyte-like cells and dendritic cells). These elements have a characteristic distribution pattern likely related to different local functional requirements.     

Analysis of the Behavior of Mitochondria in the Ovaries of the Earthworm Dendrobaena veneta Rosa 1839

We examined six types of cells that form the ovary of the earthworm Dendrobena veneta ogonia, prooocytes, vitellogenic oocytes, trophocytes, fully grown postvitellogenic oocytes and somatic cells of the gonad. The quantitative stereological method revealed a much higher “volume density” of mitochondria in all of the types of germ-line cells except for the somatic cells. Fluorescent vital stain JC-1, however, showed a much higher oxidative activity of mitochondria in the somatic cells than in the germ-line cells. The distribution of active and inactive mitochondria within the studied cells was assessed using the computer program ImageJ. The analysis showed a higher luminosity of inactive mitochondria in all of the types of germ-line cells and a higher luminosity of active mitochondria in somatic cells. The OXPHOS activity was found in somatic cells mitochondria and in the peripheral mitochondria of the vitellogenic oocytes. The detection of reactive oxygen species (ROS) revealed a differentiated distribution of ROS in the different cell types. The amount of ROS substances was lower in somatic cells than in younger germ-line cells. The ROS level was also low in the cytoplasm of fully grown postwitellogenic oocytes. The distribution of the MnSOD enzyme that protects mitochondria against destructive role of ROS substances was high in the oogonia and in prooocytes and it was very high in vitellogenic and postvitellogenic oocytes. However, a much lower level of this protective enzyme was observed in the trophocytes and the lowest level was found in the cytoplasm of somatic cells. The lower mitochondrial activity and higher level of MnSOD activity in germ-line cells when compared to somatic cells testifies to the necessity of the organisms to protect the mitochondria of oocytes against the destructive role of the ROS that are produced during oxidative phosphorylation. The protection of the mitochondria in oocytes is essential for the transfer of healthy organelles to the next generation.     

Fine structure of the parotid gland in the pika and the volcano rabbit

The parotid glands of the pika and the volcano rabbit were examined by light and transmission electron microscopy. The acinar cells of the pika consisted of light cells containing basophilic granules of low density, while in the volcano rabbit the acinar cells consisted of light and dark cells containing acidophilic granules of moderate density. Intercalated duct cells were composed of light cells containing a few granules of moderate density. These segments of the two animals were similar in morphology. The striated duct cells in both species were composed of light and dark cells. Most of those in the pika contained a few moderately dense granules. In both animals, no myoepithelial cells were detected around the acini, intercalated ducts or striated ducts, while nerve terminals were observed among the adjacent acinar cells.     

Ultrastructure of the APUD-like cells of the frog thymus gland

Within the thymus gland of the European common frog, Rana temporaria, cells with endocrine- like appearance have been found. At the ultrastructural level the most characteristic feature of their cytoplasm is the presence of secretory granules. Some cells possess irregular electron lucent granules with an eccentrically located dense core while others possess smaller electron dense granules. The cytoplasm contains also cisterns of rough endoplasmic reticulum, free ribosomes, and small mitochondria. The cells possess irregular nuclei with the pronounced nucleoli. These endocrine-like cells are connected by desmosomes with neighbouring non-granulated epithelial cells. Ultrastructural features of the cells described here resemble those seen in polypeptide hormone-secreting cells belonging to the family of cells of the APUD (Amine Precursor Uptake and Decarboxylation) series.     

The postnatal development of the submandibular gland of the mouse

Summary The postnatal development of the submandibular gland was investigated in male mice of the Swiss-Webster strain, which were killed at 1, 2, 3, 4, 5, 6, 8, 10, 12, 16 and 20 weeks of age, while the older mice had been weaned at 3 weeks of age. The mean weight of the submandibular gland increases from 9.5 mg at 1 week to 232.9 mg at 20 weeks of age, and the rate of increase is rapid between 3 and 10 weeks of age. The gland's contents of DNA, RNA and protein increase in a similar manner.The changes in the constituent cell types of the gland were studied in radioautographs prepared from Epon-embedded sections of mice given ³H-thymidine and stained with toluidine blue. At 1 week of age, the gland consists of acinar cells (36%), intercalated duct cells (26%), juxta-acinar cells (13%), striated duct cells (12%) and others. The cellular composition of the gland changes little before weaning, but the absolute number of all types of cells increases with age. Between 3 and 4 weeks, juxta-acinar cells disappear and granular convoluted tubule cells appear and increase rapidly in number with age. The rapid expansion of the population size of granular convoluted tubule cells after weaning coincides with the second peak of increased proliferative activity of intercalated duct cells, whereas all the other cell types show a progressive decrease in their proliferative activity with age. In spite of the burst in proliferative activity, there is no corresponding increase in the absolute number of intercalated duct cells. The number of striated duct cells peak at 5 weeks of age and then declines. These findings indicate that the mitoses of intercalated duct cells give rise to granular convoluted tubule cells through a stage of striated duct cells. At 20 weeks of age, the gland consists of granular convoluted tubule cells (47%), acinar cells (28%), intercalated duct cells (12%), striated duct cells (1%) and others.Supported by Public Health Service Research Grant AMDE 19753 from the National Institute of Health. The authors are indebted to Mr. I. Borcsanyi for technical assistance     

Mast cells and cells producing alpha-endorphin in the mucosa of the antral section of the stomach

The distribution of mast cells, alpha-endorphine-producing cells (AER-cells) and argyrophilic cells in lamina propria of the antral mucosa was determined quantitatively in 13 normal men. The cells were detected by histochemical and immunohistochemical (PAP) methods. The form and site of AER-cells resembled those of mast cells and mucosal argyrophilic cells (in lamina propria). The authors assume that part of human gastric mucosal cells have argyrophilic properties and contain alpha-endorphine.     

A morphologic study of the ductus of the epididymis of Sus domesticus

The head, body, and tail regions of the epididymal duct (or caput, corpus, and cauda epididymis) in two healthy and sexually mature Sus domesticus males were examined by light microscopy and by scanning or transmission electron microscopy. The epididymal duct is lined with a pseudostratified epithelium with stereocilia and covered by a muscular-connective tissue sheath that is thickest in the tail region. Diameter of the epididymal duct and height of epididymal epithelium are maximal in the head region. Length of the sterocilia and spermatic density are higher in the head and body regions. Somatic cells are abundant in the tail region. The epididymal epithelium is made up of five cell types: basal cells, principal cells, clear cells, narrow cells, and basophilic cells. Abundant secretory units are observed in the supranuclear cytoplasm of columnar principal cells. Each mature secretory unit is constituted by electron-dense secretion granules covered by more than eight layers of cisternae of reticulum between which the mitochondria are intercalated. In the apical cytoplasm the isolated secretion granules become larger and less electron dense. The apical surface is covered by numerous sterocilia. Basal cells are pyramidal and less high than principal cells. The clear cells, arranged between the principal cells, are characterized by the presence of abundant vesicular elements and electron-lucid secretion granules, and by an apocrine secretory process. The narrow cells are characterized by their highly vacuolized cytoplasm. Intermediate cell typologies can be found among basal, principal, clear, and narrow cells, which could be four developmental stages of the same cell type. The basophilic cells are spheroidal and are found at different levels between the epithelial cells and in the connective tissue underlying the epithelium. © 1993 Wiley-Liss, Inc.     

Multiplication of the dictyosome during the formation of autospores in the green alga Chlorococcum infusionum

Numerical changes in dictyosomes during the formation of autospores in a green alga, Chlorococcum infusionum, were investigated by electron microscopy. Two dictyosomes were seen near the nucleus in young vegetative cells. Four dictyosomes were seen in large mononucleate cells which appeared to enter mitosis soon. Binucleate cells contained 4 or 8 dictyosomes, the latter number being found in the large binucleate cells. Large tetranucleate cells contained 16-25 dictyosomes in each cell. Dictyosomes consisted of about 4 cisternae with a diameter of 0.4-0.6 micron in mono- to tetranucleate cells. Cytokinesis began with the formation of the septa during the third nuclear division, and 16 cells were finally formed. Dictyosomes did not increase in number in 8- and 16-nucleate cells. In septum-forming cells, dictyosomes were 0.6-1.0 microns in diameter, with 6-9 cisternae. A single dictyosome was included in each of the 16 resultant cells. These observations suggest that the dictyosomes multiply in association with the multiplication of the nuclei without correlation with formation of the cell wall or cytokinesis.