A checkerboard pattern manifested by the oviduct epithelium of the Japanese quail.

The oviduct epithelium of the Japanese quail is a monolayered epithelium consisting of two types of columnar cells, goblet type gland (G-) cells and ciliated (C-) cells. We found these cells to be arranged in a checkerboard pattern. Three types of cell boundaries formed between the two different types of cells were examined statistically at various levels of the columnar cells. There was a tendency on the part of the cells to form boundaries between G- and C- cells rather than between two C- cells or between two G- cells. We therefore propose that the pattern is constructed under a rule of maximizing the length of boundaries of two different types of cells owing to the fact that theirs is the greatest adhesion capacity. The role of microfilament bundles running along the apical cell boundaries is also discussed. It is suggested that they are in a tense state so as to shorten total length by contraction.     

Development of sperm storage tubules in the quail during sexual maturation.

The development of the utero-vaginal sperm storage tubules (SST) in the quail during sexual maturation was studied using light microscopy and image analysis. SST development starts at around 28 days with low columnar cells, 10.9 +/- 0.7 microm high, found at the base of mucosal folds in the distal uterus. Seven days later, small bud-like invaginations consisting of columnar, 16.4 +/- 1.1 microm high cells with basal nuclei were found in this region. An extremely rapid growth of the oviduct occurred at approximately 40-42 days of age with considerable variation in oviductal length between birds, coefficient of variation (CV) 64.8% and 66.2%, respectively. Two of these birds had SST containing spermatozoa but were not laying. At 49 days, oviductal length was 24 +/- 0.5 cm (CV 2.0%), and all birds had functional SST with spermatozoa and had started to lay. Mature SST consist of columnar, nonciliated cells, 19.8 +/- 0.7 microm high. Although development of SST in the quail, to a large extent, coincides with the development of the rest of the oviduct, the present findings suggest that utero-vaginal sperm storage is possible before the complete maturation of the oviduct and subsequent onset of lay.     

Heterogeneity of progesterone receptor expression in epithelial cells of immature and differentiating quail oviduct

The localization of progesterone receptor (PR) in the quail oviduct was investigated before and after the onset of sexual maturation using an immunohistochemical technique. PR was revealed exclusively in nuclei of target cells whatever the hormonal state of the tissue (immature or not, pretreated or not with progesterone). In the immature or ovariectomized quail oviduct, PR was principally localized in the undifferentiated epithelial cells; some mesothelial cells and a very few stromal cells expressed the PR. Only 40-45% of the epithelial cells were immunoreactive. These positive cells were mainly localized in the furrows of the villi where further evagination of the epithelium will occur to form the tubular glands. The onset of sexual maturation was accompanied by an increase of the proportion of positive epithelial cells and stromal cells. In estradiol-treated animals, more than 90% of the tubular gland cells were strongly stained while only 40% of the luminal epithelial cells were immunoreactive. Our results show that there are two subpopulations of epithelial cells: those expressing the PR before the onset of sexual maturation even in ovariectomized quails (constitutive expression) and those expressing the PR during sexual maturation or after estrogen injection (inductive expression). These results, associated with previously published studies dealing with the cytodifferentiation of epithelial cells during natural development or after various hormonal treatments in ovariectomized animals, suggest that the first are the progenitors of tubular gland cells, and the second the progenitors of ciliated and goblet cells. In stromal cells, PR expression is also inducuible.     

Fine structural observations on magnum mucosa in quail and hen oviducts

Summary Previous investigators have reported that albuminous material in the albumin-secreting (tubular gland) cells of the magnum of hen oviduct accumulates in the ergastoplasmic cisternae and is released directly into the glandular lumen without being first concentrated into secretory granules in the Golgi region (Zeigel and Dalton, 1962). Present fine structural studies on the tubular gland cells in oviducts from actively laying wild-type Japanese quail and in an oviduct from an actively laying hen indicate that the Golgi apparatus is directly involved in the formation of secretory granules. At least three types of granules can be observed in the tubular gland cells at various times, and all types seem to be associated initially with the Golgi apparatus.In actively laying quail, the distribution of electron opaque, intermediate, and light granules within the superficial and deep regions of the glandular epithelium varies, depending on the presence of an egg in a particular region of the oviduct. Secretion of the product is merocrine, involving fusion of granule membranes with the plasmalemma of the cell surface.Granules first appear in the tubular gland cells of quail oviducts at about 4 1/2 weeks of age. The granules are of the electron opaque type and probably represent secretion in concentrated storage form. At this age, a few of the tubular gland cells exhibit distended ergastoplasmic cisternae containing material of low electron density. The appearance of these light cells, which occur with greater frequency in oviducts from older quail, probably reflects an increased level of secretory activity initiated by changes in hormonal levels. From 5 weeks of age on, intermediate and light (less concentrated) granules, as well as dark granules, are present.Supported by the National and Medical Research Councils of Canada.     

Sex-steroid-sensitive stromal cells and oviduct differentiation

The chick oviduct differentiates during sexual maturation before the age of 20 weeks. In the present work we used immunohistochemistry to study sexual maturation associated progesterone receptor (PR) expression in the chick oviduct as an indication of progesterone sensitivity. Since the PR is estrogen inducible protein, its expression also reflects the effects of endogenous estrogens. Thus PR expression can be used as a marker for action and sensitivity of cells to these sex steroids. In the luminal epithelium and mesothelium (peritoneal epithelium) the PR was expressed in high concentrations from the time before hatching (the constitutive PR). The PR was not detectable in stromal cells of immature chicks. At the age of 7-10 weeks the PR was detected in submucosal but not in mucosal stromal cells (the inductive PR). The appearance of these PR-expressing cells was associated with an increase in luminal epithelial cell proliferation. At the age of 14-16 weeks the mucosal plicae increased in height and the PR-expressing stromal cells were seen in the center of these mucosal plicae. There were also areas in the mucosal plicae where a large number of stromal cells expressing the PR were seen in the mucosal layer. Thereafter the size of the oviduct increased rapidly and the gland formation commenced. In the fully matured oviduct (over 18 weeks of age) virtually all stromal cells both in mucosa and submucosa expressed the PR. It is concluded that the PR expression in the luminal epithelium and mesothelium was constitutive (independent of sexual maturation). In stromal cells this was expressed during sexual maturation (probably induced by endogenous estrogen) and was associated with histological changes in the oviduct. We propose that direct effects of estrogen and progesterone in the oviduct growth and glandular formation are mediated through these stromal cells.     

Postnatal development of the oviduct of intact and ovariectomized rabbits

The length of the oviduct, the thickness of its wall, and the height of its mucosal epithelium and cilia were measured in (a) 0-, 2-, 4- and six-month-old rabbits, (b) rabbits ovariectomized at birth and (c) ovariectomized, estrogen-treated rabbits. The length and external diameter of the oviduct increased progressively until four months of age, after which their rates of increase declined. The thickness of the oviductal wall at the uterotubal junction was twice as large as that of the isthmus at two months of age and six times as large at four and six months of age. The height of the mucosal epithelium in the fimbriae was less than that in other oviductal segments at birth, but exceeded that in other segments at six months of age. Ciliated cells and motile cilia were absent 24 hours after birth; they were first observed two months after birth. The cilia of fimbriae were shorter than cilia elsewhere in the oviduct. Neonatal ovariectomy retarded the development of the oviduct and the mesotubarium and caused pyknosis of ciliated and non-ciliated cells of the oviductal mucosa. Cells with scarcely motile cilia were present five and one-half months after neonatal ovariectomy.     

Determination of ciliary polarity precedes differentiation in the epithelial cells of quail oviduct.

In quail oviduct epithelium, as in all metazoan and protozoan ciliated cells, cilia beat in a coordinated cycle. They are arranged in a polarized pattern oriented according to the anteroposterior axis of the oviduct and are most likely responsible for transport of the ovum and egg white proteins from the infundibulum toward the uterus. Orientation of ciliary beating is related to that of the basal bodies, indicated by the location of the lateral basal foot, which points in the direction of the active stroke of ciliary beating. This arrangement of the ciliary cortex occurs as the ultimate step in ciliogenesis and following the oviduct development. Cilia first develop in a random orientation and reorient later, simultaneously with the development of the cortical cytoskeleton. In order to know when the final orientation of basal bodies and cilia is determined in the course of oviduct development, microsurgical reversal of a segment of the immature oviduct was performed. Then, after hormone-induced development and ciliogenesis, ciliary orientation was examined in the inverted segment and in normal parts of the ciliated epithelium. In the inverted segment, orientation was reversed, as shown by a video recording of the direction of effective flow produced by beating cilia, by the three-dimensional bending forms of cilia immobilized during the beating cycle and screened by scanning electron microscopy, and by the position of basal body appendages as seen in thin sections by transmission electron microscopy. These results demonstrate that basal body and ciliary orientation are irreversibly determined prior to development by an endogenous signal present early in the cells of the immature oviduct, transmitted to daughter cells during the proliferative phase and expressed at the end of ciliogenesis.     

Evolutionary conservation of an epitope associated with striated rootlets in different epithelial ciliated cells.

In ciliated cells of metazoa, striated rootlets associated with basal bodies anchor the ciliary apparatus to the cytoskeleton. We have used here a monoclonal antibody against a 175 kDa protein associated with the striated rootlets of quail ciliated cells, to study ciliated cells of different species. In mussel gill epithelium the antibody recognized a protein of 92 kDa which shows a periodic distribution along the striated rootlets. In frog ciliated palate epithelium, two different rootlets are associated with basal bodies, both are decorated and only one protein of 48 kDa is recognized on immunoblot. The antigen is arranged in a helix around the striated rootlets. In rabbit ciliated oviduct epithelium, we detected the presence of very small and thin rootlets which are weakly labeled. We have shown that an epitope associated with the striated rootlets is preserved through evolution although the molecular weight of the peptide varies. We have also observed the appearance of this epitope on protein associated with junctional complexes in rabbit and cytoskeleton component in quail oviduct.     

First evidence of the interaction between deleted in malignant brain tumor 1 and galectin-3 in the mammalian oviduct

The oviduct supports the transport and final maturation of gametes, and harbors fertilization and early embryo development. The oviductal epithelium is responsible for providing the correct environment for these processes. Deleted in malignant brain tumor 1 (DMBT1) is expressed by multiple organisms and several cell types, and the interaction of the rabbit ortholog of DMBT1 with galectin-3 (gal-3) modulates the polarity of epithelial cells. This interaction has not yet been shown in locations other than rabbit kidney and human-cultured endothelial cells. DMBT1 and gal-3 also protect epithelial layers from pathogens and trauma, and are innate immunity components. DMBT1 has been detected in the porcine oviduct, and gal-3 has been reported in the Fallopian tube and in the cow oviduct. Interaction between both proteins would show a probable physiological function in the female reproductive tract. This work describes the presence and co-localization of DMBT1 and gal-3 mainly in the apical region of the epithelial cells of the Fallopian tube and the porcine oviduct, and co-immunoprecipitation in membrane-enriched epithelial cell extracts from the porcine oviduct. The findings strongly support a functional interaction in the mammalian oviduct, suggestive of a role on epithelial protection and homeostasis, which might be related to epithelium–gamete interaction.     

Zinc Secretion in the Oviduct of the Coturnix Quail

The oviduct from laying quail were used to investigate mechanisms of trace mineral secretion and the possible role of metallothionein in this process. Secretion of zinc occurred maximally at pH 5.4, which is close to the normal pH of the oviduct. Secretion occurred to a much greater extent in the isthmus and shell gland than in the magnum, the major protein-secretory section of the oviduct. Intraperitoneal administration of cadmium resulted in a marked reduction in Zn secretion from the oviduct of laying quail. This effect could not be correlated with metallothionein since metallothionein could not be detected in any section of the oviduct in control or Cd-induced quail. Small-molecular-weight metal-binding ligands were present in the isthmus and shell gland, which may play a role in trace mineral mobilization. Histological evaluation by light and elelctron microscopy show that Zn is transported from the smooth muscle cells through the connective tissue matrix in the extracellular space to the epithelial goblet cells. Presumably, Zn and other trace minerals are secreted from the secretory goblet cells into the egg.     

Histometry of normal thyroid glands in neonatal and adult rats

The present histometric study is on thyroid glands of Wistar rats ranging in age from 0 to 120 days. The mean volume of one lobe of the thyroid in 4-month-old animals was some 22-, 10-, 5-, and 3-fold greater, respectively, than the volumes in the newborn, 5-, 10-, and 30-day-old rats. At 4 months of age the mean length of the lobe was 3 times greater than at birth. The volumetric fractions (Vv) of the different histological components (follicular cells, C-cells, colloid, and interstitial tissue) changed considerably in the course of development. The Vv of follicular cells diminished from 61.4% at birth to 37.2% at 4 months. C-cells increased from 2.9% in the newborn to 4% at 15 days, with no further significant change at 4 months. Colloid and stroma together represented 35.7% at birth, increasing to 58.5% at 120 days. In the course of the first 4 months of life, the absolute volumes occupied by follicular cells, C-cells, colloid, and stroma increased 13.25, 30.75, 38.6, and 33.7 times, respectively; these changes reflect the variations that occur in the volume of the gland and the Vv throughout postnatal development.     

DEVELOPMENT OF VAGINAL EPITHELIUM SHOWING IRREVERSIBLE PROLIFERATION AND CORNIFICATION IN NEONATALLY ESTROGENIZED MICE: AN ELECTRON MICROSCOPE STUDY *

Irreversible proliferation and cornification of the mouse vaginal epithelium were induced by 10 daily injections of 20μg estradiol-17β starting on the day of birth. Development of the irreversible vaginal epithelium during the period of estrogen injections in early postnatal life was observed under light and electron microscopes. Small electron-dense cells (A-cells) in clusters were present in the columnar vaginal epithelium of newborn mice. A-cells were proliferated by 2 daily estrogen injections. At the sites of A-cell clumps, large electron-dense cells (B-cells) characterized by long winding cytoplasmic processes appeared in mice given 3 daily injections, forming nodules which then fused together to form a layer under the columnar epithelium after 4 daily injections. In mice given 7 daily injections, the primary epithelium was shed by the superficial cornification of the newly formed layer. The B-cell membrane bore fewer desmosomes than in the basal and intermediary cells of the vaginal epithelium of ovariectomized ‘normal’ adult mice after 5 daily injections of 100μg estradiol-17β. Hyperplastic epithelial downgrowths in old ovariectomized mice given neonatal estrogen injections contained another type of cells with reduced density which formed much fewer processes and only a few desmosomes (C-cells).     

Immunohistochemical study of the C-cell complex of dog thyroid glands with reference to the reactions of calcitonin,C-thyroglobulin and 19S thyroglobulin

Summary Continued from the previous study in fetal animals (Kameda et al. 1980), the development and maturation of C-cell complexes in postnatal dogs from newborn to adult were investigated by use of an immunoperoxidase method using antisera to calcitonin, C-thyroglobulin (C-Tg) and 19S thyroglobulin, respectively. The younger the animals were, the more numerous were undifferentiated cells and high columnar epithelial cells in the complexes. With increasing age, the constituent elements of the complexes progressively differentiated. In one type of complex there are a large number of C-cells in various developmental stages, as well as undifferentiated cells and cysts. C-cell complexes composed mostly of mature C-cells were regarded as the more highly differentiated structures of this type. A second type contains follicular cells in various stages of differentiation in addition to undifferentiated cells and C-cells, i.e., 19S-positive cell masses not yet organized into follicles, primordial follicles with small lacunae and comparatively larger follicles. The follicular cells in the complexes were similar with respect to immunoreaction and folliculogenesis to the cells of fetal thyroids, but they developed very slowly. In conclusion, the present study indicates that follicular thyroid cells can differentiate within C-cell complexes, i.e., they develop from cells of ultimobranchial body origin.     

Localization of immunoreactive gonadotropin-releasing hormone and relative expression of its mRNA in the oviduct during pregnancy in rats.

This study was designed to determine the cellular and ultrastructural distribution of the gonadotropin-releasing hormone (GnRH) and the relative expression of its mRNA in the oviduct of rats during different time points (days 7, 9, 16, and 20) of pregnancy. Immunofluorescent localization and confocal microscopic techniques were used to determine the cellular distribution of GnRH in the oviduct. Immunogold electron microscopy indicated its localization at the ultrastructural level, and real-time PCR was used to study the expression pattern of GnRH mRNA in the oviduct during pregnancy. In general, GnRH was localized within the epithelial cells lining the oviductal lumen at each selected time point. A strong correlation between the fluorescence intensity of GnRH-immunoreactive cells and the relative expression of GnRH mRNA was noted on days 7 and 16, followed by a plateau by day 20. At the ultrastructural level, uniform labeling of colloidal gold particles was observed in secretory vesicles and lamella of the luminal epithelium as well as the lumen of the oviduct. Collectively, these results demonstrate for the first time that the oviductal epithelium synthesizes and secretes the decapeptide GnRH during pregnancy in rats, which may have a possible role in postimplantation embryonic development and the maintenance of pregnancy.     

An ultrastructural and histochemical study of the development of secondary palate in Japanese quail, Coturnix coturnix japonica

Embryonic development of the secondary palate of the Japanese quail, Coturnix coturnix japonica, was studied. The palatal shelves appeared on day 5 of incubation in a horizontal direction over the dorsal surface of the tongue. Subsequently, unlike those in mammals, the opposing palatal shelves do not fuse, and a physiological cleft persists between them. In comparison with the chick, where the palatal shelves do not fuse and the medial edge epithelium (MEE) becomes orthokeratinized stratified squamous type, the quail MEE differentiates into a parakeratinized stratified squamous type. The basal cells of the quail MEE differentiated from cuboidal to columnar and showed reorganization of their cytoplasmic organelles. In contrast to both the chick and mammalian MEE, quail MEE showed very little ruthenium red (RR) binding at the plasma membrane of both the basal and superficial cells. Initial development in a horizontal direction, lack of fusion, and an absence of programmed cell death in MEE of developing quail palate distinguished it from the mammalian palatogenesis. Also, although the morphogenesis of palate in quail and chick was similar, the pattern of cytodifferentiation in their MEE was different, which may be attributed to their ontogenesis.     

Survival in foods of Staphylococcus aureus grown under optimal and stressed conditions and the effect of some food preservatives

Staphylococcus aureus was grown in a rich peptone medium which became alkaline with continued incubation. Cells were grown at 37 degrees C and in the same medium containing 1 M NaCl at 46 degrees C, a temperature at which this organism can grow only when protected by NaCl. Cells of these cultures are hereafter called 37 degrees C-cells and 46 degrees C-cells, respectively. The 37 degrees C-cells harvested when the pH was 7.1 to 7.7 had decimal reduction times (D60-value) of 1.8 to 3.1 min in 50 mM pH 7.2 Tris buffer. The D60 value of 46 degrees C-cells tested in the same way, harvested from cultures at pH 6.6 to 7.6, ranged from 5.3 to a maximum of 12.8 min. In milk, green beans, peas, or beef slurry, the D60-value of 46 degrees C-cells was about four times higher than that of 37 degrees C-cells. Length of survival after freeze-drying in skim-milk powder exposed to air was longest for the cells with the highest D-value. In freeze-dried peas and media acidified with acetic and lactic acids, 46 degrees C-cells survived longer than 37 degrees C-cells. However, the sensitivity of the two kinds of cells to potassium sorbate, sodium benzoate, and sodium propionate was essentially the same, but the 46 degrees C-cells were more resistant to butylated hydroxyanisole and sodium nitrite.     

Morphology, histochemistry and biochemistry of the oviduct of the toad, Bufo vulgaris (Anura, Amphibia), during the annual reproductive cycle.

On the basis of histological and histochemical characteristics the oviduct of adult Bufo vulgaris can be separated into six zones. These characteristics include mainly the relative abundance of secretory and ciliated epithelial cells, mucosal foldings, tubular glands and the staining properties of the secretory elements. The VI zone (uterine segment) is totally deprived of tubular glands and is prodoundly rich in epithelial secretory cells. These cells and the tubular glands, when well-developed, are rich in neutral and acid mucins in the preovulatory phase, whereas this content is greatly diminished after ovulation. Similarly, the weight and the length of the oviduct displays a marked seasonal pattern. The same is observed for its protein content and acid and alkaline phosphatases. These parameters show the highest values in the preovulatory period and lowest after ovulation. The toad oviduct also displays marked hydroxysteroid dehydrogenase activity along the mucosal epithelium. Its functional significance remains a matter of debate, however. A marked parallelism between the present data and those obtained by others in other amphibians indicates the existence of a common pattern of seasonal modifications correlated mainly with the release of the eggs and the formation of their multi-layered coat.     

Reactivity of monoclonal islet cell antibodies on normal hyperplastic and neoplastic thyroid C-cells

Summary Thyroid C-cell reactivity to 15 monoclonal antibodies raised against a series of pancreatic islet cells (H[human]ISL, B[bovine]ISL and R[rat]ISL) was evaluated using an indirect immunoperoxidase technique on frozen thyroid sections. Of the monoclonal anti-islet cell antibodies, five reacted specifically with bovine C-cells or human hyperplastic and neoplastic C-cells but not with follicular cells. Two monoclonal antibodies of the bovine series showed strong immunoreactivity with C-cells and only a weakly positive immunostaining of follicular cells. Five monoclonal antibodies reacted with both thyroid C-cells and follicular cells, whereas 3 monoclonal anti-islet cell antibodies did not stain any cell type of the thyroid. In human medullary carcinomas, calcitonin- and somatostatin-producing neoplastic cells were immunoreactive with the same monoclonal antibodies as were normal human C-cells. The protein bands identified by the monoclonal antibodies in human medullary carcinomas had the same molecular weight as those from pancreatic islet extracts. Our study demonstrates the presence of similar differentiation antigens on thyroid C-cells and pancreatic islet cells; this further illustrates common modes of differentiation and specialisation of these embryologically different members of the dispersed neuroendocrine system. The crossreactivity of seven of the monoclonal antibodies investigated with follicular epithelium of the thyroid suggests the existence of common antigenic determinants in different endocrine organs and may partly explain the multiple organ autoimmune response found in patients with polyendocrine diseases.