Development of thyroid gland and ultimobranchial body cyst is independent of p63.

The ultimobranchial body (UBB) and thyroid primordium are the origins of the thyroid gland that fuse around embryonic day 14.5 of mouse gestation, ultimately giving rise to calcitonin-producing C cells and thyroglobulin-producing follicular cells, respectively. A homeodomain transcription factor NKX2-1 is expressed both in the UBB and the thyroid primordium, and is critical for development of the thyroid gland. In this study, the role of p63 in development of UBB and the thyroid gland was analyzed by histological, immunohistochemical, and electron microscopic analyses using mice with various combinations of Nkx2-1 and p63 wild-type, heterozygous, and null alleles. In the absence of p63, a normal thyroid gland develops, as revealed by expression of thyroglobulin and calcitonin, thus showing that p63 is not required for thyroid development. However, in mice carrying the Nkx2-1-null allele, the UBB remains as a cystic vesicular structure and/or in nested patterns consisting of p63-positive cells surrounding the vesicle and undifferentiated immature cells with occasional cilia lying inside. The cystic UBB was present even in the Nkx2-1;p63 double-null mice. The structure and p63 expression pattern of the UBB cyst strikingly resemble the solid cell nest. These results show that in the absence of NKX2-1, UBB becomes cystic independent of p63, which is likely the origin of SCN.

Fine structure of the parathyroid and ultimobranchial glands of the snake, Elaphe quadrivirgata.

The ultrastructure of the parathyroid gland and ultimobranchial gland of the snake, Elaphe quadrivirgata, was studied. The parenchyma of the parathyroid gland was consisted of chief cells arranged in cords. Oxyphil cells and water-clear cells were not recognized. The chief cells contained irregular shaped nucleus, moderately-developed mitochondria, granular endoplasmic reticulum and Golgi complexes. A few secretory granules of 100-300 nm in diameter distributed in the cytoplasm. Small aggregations of glycogen particles and lipid droplets were dispersed widely in the cytoplasm. The ultimobranchial glands consisted of follicles and interfollicular aggregates of C-cells. Follicles were invaginated and composed of C-cells, goblet cells and ciliated cells. C-cells were located in the basal position of the follicle and possessed variety of secretory granules of 100-300 nm in diameter in the basal region of the cell. C-cells presented various secretory cycles. Goblet cells projected microvilli into follicular lumen. At the apical region the goblet cells had large granules of 300-1,200 nm in diameter. Ciliated cells projected cilia intermixed with micovilli from the apical surface of the cell into the follicular lumen.

Morphological and functional aspects of reptilian ultimobranchial gland.

The intention of this review is to compare studies on the morphology and histology (light and electron microscopic) of ultimobranchial glands of various groups of reptiles. Moreover, experiments (including our investigations) on suppression or stimulation of the ultimobranchial gland are included. Adult reptiles possess one (on the left side) or two ultimobranchial glands (UBG). The UBG lie just anterior to the heart. Light as well as electron microscopically, the gland has been shown to contain follicles and cell cords (cell aggregates). The follicular epithelium is lined by simple cuboidal or pseudostratified columnar cells. Ciliated and goblet cells may be present in the follicular epithelia in some groups. The lumen may contain a colloid-like substance with desquamated cells or debris. The UBG of reptiles seem to be an active secretory organ with influence on calcium regulation. Other functions of calcitonin have also been suggested in reptiles for example in neurotransmission, in volume regulation, phosphate balance and promotion of bone calcification (at least in juveniles).

Immuno-electron-microscopic localization of enkephalin in the secretory granules of C cells in the chicken ultimobranchial glands.

In the chicken, enkephalin-immunoreactive cells and nerve fibers are distributed in the ultimobranchial glands, which consist of C-cell groups and cyst structures. Ultrastructural features of the enkephalin cells and nerve fibers were examined by immuno-electron microscopy using both the streptavidin-biotin-peroxidase method and the protein A-colloidal gold method. Immunoreactivity for enkephalin was located on the secretory granules of C cells. In 1-day-old chickens, three types of C cells were distinguished on the basis of their granule size. Type-I cells were filled with large secretory granules (200-600 nm in diameter). These elements represented a majority of the C-cell population. Type-II cells contained medium-sized granules (100-280 nm in diameter). Type-III cells displayed small secretory granules (60-200 nm in diameter). The latter cells were elongate or irregular in shape and frequently extended cytoplasmic processes into the connective tissue stroma or contacted other C cells. Enkephalin-immunoactivity was revealed by dense deposits of immunogold particles on the secretory granules of type-II and type-III cells. There were only a few type-I cells showing immunoreactivity for enkephalin. A double immunogold labeling procedure demonstrated that calcitonin and enkephalin were colocalized in the same secretory granules of type-I and type-II cells. Type-III cells were devoid of immunoreactivity for calcitonin. Enkephalin-immunoreactive nerve fibers were characterized by the presence of granular vesicles, 60-160 nm in diameter, and frequently established direct contact with the surface of C cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Occurrence of calcitonin-positive C cells within the distal vagal ganglion and the recurrent laryngeal nerve of the chicken.

The chicken ultimobranchial glands are richly supplied with nerve fibers originating from both the main trunk of the vagus nerve and its branch--the recurrent laryngeal nerve. C cells immunoreactive for calcitonin were invariably found in the large nerve bundles distributed throughout the ultimobranchial glands. In addition, these cells were often present within the distal vagal ganglia and the recurrent laryngeal nerves. The frequency of occurrence and the pattern of distribution of the C cells in the distal vagal ganglia and the recurrent laryngeal nerves were determined in chickens of various ages by means of an immunoperoxidase method with anticalcitonin and antineurofilament antisera. The left and right sides of the ultimobranchial region were asymmetrical. The left ultimobranchial gland was in close contact with the vagus nerve trunk, especially with the distal vagal ganglion, but it was separated from the recurrent laryngeal nerve. The right gland contacted the recurrent laryngeal nerves, its medial edge being frequently penetrated by the nerve, but the gland was separated from the distal vagal ganglion. On the left side, C cells were found in 25 out of 39 distal vagal ganglia but they were not distributed in the recurrent laryngeal nerve. On the right side, the cells were present in 28 out of 43 recurrent laryngeal nerves but absent in the distal vagal ganglia. The results indicate that the C cells secreting a hormone calcitonin can enter into nerves, but their occurrence is restricted to the nerves in close proximity to the ultimobranchial glands. Electron microscopic studies revealed that C cells in the nerves received numerous axon clusters enveloped with Schwann cell cytoplasm. Naked axons regarded as axon terminals were found in direct contact with the surface of C cells. They were mainly composed of efferent-type nerve endings showing the accumulation of numerous small clear vesicles and a few large dense-cored vesicles. In addition, C cells were partly covered with the long cytoplasmic processes of Schwann cells and were also in contact with the Schwann cell perikarya. The C cells in nerves appear to be controlled by neural stimulation.

Ultimobranchial body and parathyroid gland of the frog, Rana tigrina in response to calcitonin administration.

The effects of salmon calcitonin (0.25 MRC mU/g body wt) on the serum calcium and phosphate levels as well as on the activity of ultimobranchial body and parathyroid glands was investigated in the frog, Rana tigrina for 15 days. The hormone evokes hypocalcemia (on day 1 and day 3) which is followed by a significant hypercalcemia on day 10. Thereafter, the level of calcium decreases again on day 15. Calcitonin induces hypophosphatemia (on day 3 and day 5). Thereafter, hyperphosphatemia is recorded on day 10. By day 15 normal serum phosphate value is achieved. After treatment with calcitonin, the ultimobranchial body becomes inactive and the parathyroid glands get activated.

Localization of immunoreactive keratins in cyst epithelium of chick ultimobranchial glands.

The cyst structures of chick ultimobranchial glands were studied by electron microscopy and immunocytochemistry to characterize the type of intermediate-sized filaments present in the cells lining cyst lumina. Electron microscopy showed that the majority of the lumen-bordering cells contained extensive meshworks of intermediate-sized (7-11 nm) filaments, many of which were arranged in bundles. Apical regions of C cells directly bordering on cyst lumina were also filled with thinner (5-6 nm) filaments. Immunoperoxidase staining showed that the majority of cyst epithelial cells were stained intensely with anti-keratin antiserum, but not with anti-neurofilament antiserum, which is a specific marker for neuronal differentiation. The cyst epithelium also showed moderate-to-weak immunoreactivity for actin. Subsequently, the differentiation and maturation of cyst structures related to intermediate filament expression were studied. In 18-day-old chick embryos, keratin immunoreactivity began to appear in the cell clusters destined to form cysts and in the primordial cysts with small cavities. At this time, fine networks of intermediate filaments were already detected in the cells lining the cystic cavities. At 1 day after hatching, the cysts became a consistent feature of ultimobranchial glands. Intermediate filaments associated in bundles were observed, and the intensity of immunostaining for keratins increased. Thereafter, with progressive enlargement of cysts, numbers of intermediate filaments and intensity of keratin immunoreactivity gradually increased with age. Thus, the data indicate that in cyst epithelium keratin filaments are highly organized and may confer the structural strength necessary for cells lining cyst lumina.

The role of ultimobranchial bodies in the modulation of the response of chick embryos to 1,25-dihydroxy-cholecalciferol.

Ultimobranchial bodies (UBBs) were dissected from 17-day-old chick embryos and grafted onto the chorioallantoic membrane of 8-day-old embryos. The embryos with UBB grafts as well as sham-grafted controls were injected on the 10th day of incubation with 100 ng 1,25(OH)2D3 dissolved in ethyl alcohol or with an equal volume of ethyl alcohol alone; embryos were sacrificed on the 13th day. Grafted UBBs showed ultrastructural characteristics typical of actively secreting glands. A histological study of the tibiae from all embryos showed that while the grafted embryos responded to the injection of 1,25(OH)2D3 with a peripheral rim of undermineralized bone trabeculae, sham-grafted embryos never did so. These results confirm the original hypothesis that the presence of differentiated UBBs is a precondition for the production of undermineralized bone (osteoid) by 1,25(OH)2D3. In a second series of experiments, similarly treated embryos were sacrificed on the 10th, 11th, 12th and 13th day; the levels of calcium and inorganic phosphate were determined in their blood. The injection of 1,25(OH)2D3 produced in all embryos hypercalcaemia and hypophosphataemia. However, the hypophosphataemic response was more prolonged in the embryos with UBB grafts than in sham-grafted ones. These results suggest that the grafted UBBs prolonged the hypophosphataemic response, probably by secreting calcitonin and thus reducing the rate of bone resorption. It is also probable that the prolonged hypophosphataemia produced or contributed to the undermineralization of the peripheral (subperiosteal) trabeculae.

An ultrastructural study of the cysts in chicken ultimobranchial glands, with special reference to C-cells.

The ultimobranchial glands of the chicken were examined by electron microscopy and immunocytochemistry using a calcitonin antiserum. Electron microscopy confirmed the presence of C-cells, containing numerous secretory granules storing calcitonin, in the luminal lining of cyst-like structures found in these glands. These cells were furnished with prominent microvillar projections at their luminal surface, and the cytoplasm of the apical region was filled with fibril material. Furthermore, the cells contained prominent junctional complexes and desmosomes at their apico-lateral surfaces. In these C-cells, secretory granules were concentrated near the lumen and some were attached to the apical cell membrane. The luminal content of the cysts had a colloid-like and flocculent appearance, and was frequently seen attached to the cytoplasmic projections or apical cell membrane of the C-cells. Since the cysts progressively increase in volume and number with age, it is suggested that they may partly play a role in the storage of excess or unneeded hormonal products.

Ultrastructural localization of immunoreactive calcitonin in the two cell types of the ultimobranchial gland of the common toad (Bufo bufo L.).

The ultimobranchial (UB) glands of the common toad Bufo bufo consist of several cellular masses containing two quite different cell types which line a central lumen filled with amorphous material. The morphologically defined Type I cell is akin to a typical calcitonin secretory cell as observed for all vertebrates, with small dense-core secretory granules. On the contrary the Type II cell displays large apical dense bodies which may be related to the secretion and/or absorption of the amorphous material. Cells morphologically related to Type II cells have been described in the UB glands of Sauropsidea and in the UB follicles of mammalian thyroid gland. An immunocytochemical stain using an antiserum raised against synthetic salmon calcitonin demonstrated the specific localization of an immunoreactive product in both Type I and Type II cell granules, suggesting that both cell types could be involved in calcitonin metabolism. Moreover, the presence of immunoreactive calcitonin in the Type II cells of the Bufo UB gland raises the question of the function of the morphologically equivalent cells of other species.

[Ultrastructural modifications in the thyroid glands of mice treated with lithium gluconate]. / Modifications ultrastructurales de la thyroïde de Souris traitée au gluconate de lithium.

The various morphological and functional changes produced by lithium gluconate treatment of Mouse thyroid and ultimobranchial cells show complex interactions, involving direct and indirect effects. In most of the cases these effects determine metabolism (thyroid cells) or differentiation (ultimobranchial cell) disturbances. In most of the cases analysed changes do not concern the whole gland and should not have important consequences on its general activity-so it is more likely that at low concentration the effects of lithium gluconate on the thyroid gland of manic depressive patients might be neglected at the level of the functional state of the whole gland.

[Seasonal variations of the ultimobranchial body of the turtle (Pseudemys scripta)]. / Sulle modificazioni cicliche stagionali del corpo ultimobranchiale di tartaruga (Pseudemys scripta).

Ultrastructural seasonal cyclic aspects of ultimobranchial body (C.U.B.) in fresh turtles (Pseudemys scripta) are studied. The C.U.B. consists of follicles and cords. The cord cells are characterized by many secretory granules measuring approximately 180 nm with variable feature and electron density. These granules are localized in the cytoplasm close to basal laminae. The follicular cells, on the contrary, present few and large secretory granules, glycogen particles, bundle of filaments and a scarcely developing Golgi apparatus and granular endoplasmic reticulum. The apical and follicular cytoplasmatic membrane is provided with small number of cilia and short microvilli. Seasonal cellular variations are described clearly in the winter (february-march) specimens of ultimobranchial body, these are characterized by more cord aspects and few follicles. The only cord cells present significant ultrastructural changes, represented by more glycogen particles, middle and wide lipid droplets and poor presecretory granules in Golgi zone. These morphological elements orient to a parallelism with the C cells of hibernant animals (Azzali 1967, Frink and Coll.) and the chief cells of parathyroid gland of the self turtle species as by our previous study.

The carotid body of the pheasant (Phasianus colchicus L.). An electron microscopy study.

In the pheasant, the carotid body lies laterally to the common carotid artery, is partly surrounded by parathyroid tissue and is immediately adjacent to the ultimobranchial body. In some cases these tissues intermingle. The cellular component of the carotid body consists primarily of granular endocrine cells, which are arranged in islands and lie in immediate proximity to wide capillaries and nerve fibres. A quantitative evaluation of granule size frequencies in these main cells, which we divided into three types, showed, inter alia, that the types represented three stages of cell differentiation, type III being the most mature cells. Only this last, i.e. mature, type can be used for differentiation from the granular cells of the ultimobranchial body of the same species, from which they differ. In addition, the main carotid body cells are typically surrounded by sustentacular cells. Numerous contacts with the terminal parts of axons were found on the main cells.

Fine structure of the ultimobranchial body of the pheasant (Phasianus colchicus L.).

The pheasant's ultimobranchial body is characterized by the presence, in the connective tissue stroma, of epithelial cells between which two types of granulated cells comprising the main part of the glandular portion of the body are formed. Type I is characterized chiefly by the presence of nonrounded electron-dense secretory granules measuring 65-240 nm in the cytoplasm and by a very well-developed Golgi apparatus. The cytoplasm of type II cells also contains dark secretory granules, but somewhat smaller (50-150 nm). This type is very frequently in close contact with specific and often branching tubular structures from whose cells microvilli project into the lumen. The dark cytoplasm of the cells lining these structures contains a relatively large number of mitochondria and dense bodies, but no secretory granules.