Ultimobranchial Research Articles

Dynamics of plasma Ca and calcitonin levels in stonefish (Inimicus japonicus) administered a high-Ca solution into the stomach.

In stonefish, changes in plasma total Ca and calcitonin levels were examined after administration of a high-Ca solution into the stomach. Blood was taken successively at 0, 1, 3, 9, 33, and 81 hr from a fine tube cannulated into the aortic bulb. Plasma Ca levels increased acutely at 1 hr and attained the peak after 3-9 hr of the administration. Although plasma calcitonin levels did not exhibit conspicuous changes for 1-3 hr, they began to rise significantly at 33 hr. The plasma Ca level began to decline significantly at 33 hr, although the level was still significantly higher than the initial level. At 33 hr, however, the plasma calcitonin level still continued to increase. At 81 hr, the plasma Ca level had returned to the initial level. At that time, the plasma calcitonin level was also significantly lower than that at 33 hr. These results suggest that, in stonefish, the ultimobranchial gland has the ability to respond physiologically to rises in plasma Ca levels, to secrete calcitonin, and to cease the secretion when the plasma Ca levels return to the initial level.

Additional phenotypic abnormalities with presence of cysts within the empty thyroid area in patients with congenital hypothyroidism with thyroid dysgenesis.

Congenital hypothyroidism (CH) is most frequently caused by thyroid developmental abnormalities, and it has recently been shown to have a familial component with members affected by either CH or asymptomatic thyroid developmental abnormalities. The pathogenesis of the disease is unknown, but it seems possible that a common genetic mechanism underlies these heterogeneous phenotypic expressions. Associations among these anomalies in the same individuals have occasionally been described. The aim of this study was to investigate whether cysts of the thyroglossal duct could be shown by ultrasonography in patients with CH caused by thyroid dysgenesis. Children with CH (n = 57) who were diagnosed by newborn TSH screening were prospectively evaluated by ultrasonography at the age of 10.5 +/- 4.5 yr. The etiology of CH (ectopic thyroid tissue, n = 42; athyreosis, n = 15) was established before treatment initiation on the basis of thyroid radioiodine scanning and the absence of any thyroid tissue in the normal location confirmed by ultrasonography. Cysts were found in 39 patients (68% of cases) with either ectopic thyroid tissue (n = 29) or athyreosis (n = 10). All cysts were located in the empty thyroid area in the left (57%) or right (43%) side and were mostly closer to the midline. Patients had either a single cyst (n = 16 patients) or multiple cysts (n = 23 patients). The cysts were bilateral in 17 of the 39 patients. Most of them were vertically oval or round, with a size ranging in diameter from 2-21 mm (mean, 3.5 +/- 2). In conclusion, the presence of cysts within the empty thyroid area in 68% of patients with CH due to thyroid dysgenesis is a novel observation that is part of the developmental anomaly of this disease. Several explanations can be put forward to explain the presence of these cysts. They might be due to the persistence of the ultimobranchial bodies as a cystic structure or part of the thyroid-forming material, which may migrate along the normal pathway of the usual course of the thyroglossal duct, giving rise to cell residues within the empty thyroid area.

Is calcitonin an important physiological substance?

That calcitonin (CT) at supraphysiological doses is hypocalcemic led to the mistaken conclusion that it was important for calcium homeostasis and this idea has persisted to this day. Despite these findings, there is no readily apparent pathology due to CT excess or deficiency and there is no evidence that circulating CT is of substantial benefit to any mammal. Mammalian CT at physiological doses is not essential and very likely the CT gene has survived because of the gene's alternate mRNA pathway to produce calcitonin-gene-related peptide found in neural tissues. CT is not involved in calcium homeostasis or any other important physiological function, except, possibly, as an adjunct to protection of the skeleton under conditions of calcium stress, and appears to be in the process of becoming vestigial. CT produced in other tissues has paracrine actions that modulate functions such as proton transport, acid-base balance, prolactin secretion, and gastrointestinal motility. C-cells in mammals evolved from the ultimobranchial body that secretes CT in all lower vertebrates. It is highly probable that changes in amino acid sequence during evolution are responsible for the loss of activity, as fish CT is about 40 times as potent as human CT. CT may have been very important to survival in seawater fish, but the presence of the parathyroid gland and other evolutionary changes occurring in tetrapods suggest that the function of CT is no longer important.

Carotid body and glomus cells distributed in the wall of the common carotid artery in the bird.

In the bird the carotid body is located between the distal (nodose) ganglion of the vagus nerve and the recurrent laryngeal nerve at the beginning of the common carotid artery, that is, the organ is located at the cervicothoracic border. The chicken carotid body receives numerous branches from the vagus and the recurrent laryngeal nerves. In addition, dense networks of the peptidergic nerve fibers immunoreactive for substance P, calcitonin gene-related peptide (CGRP), vasoactive intestinal peptide (VIP), galanin, and neuropeptide Y (NPY) are distributed in and around the carotid body parenchyma. The substance P- and CGRP-immunoreactive fibers are derived from both the superior and inferior ganglia of the vagus nerve. The VIP-, galanin-, and NPY-immunoreactive fibers originate from the 14th cervical ganglion of the sympathetic trunk. The endocrine organs including the thyroid gland, parathyroid glands, carotid body, and ultimobranchial gland are situated as a continuous series along the common carotid artery. The organs are supplied with arteries arising as one trunk from the common carotid artery. Glomus cells are widely distributed not only in the carotid body but also in the wall of the common carotid artery and around the common trunk and its branches. The glomus cells of the chicken carotid body exhibit intense immunoreactivity for serotonin, tyrosine hydroxylase, and chromogranin A. The cells located in the wall of the common carotid artery further express NPY mRNA and peptide. In the chickens exposed to isocapnic hypoxia for 35 days, 3-4-fold increase of the carotid body volume is induced and the carotid body glomus cells show enhanced synthetic and secretory activities. On the other hand, the cells in the wall of the common carotid artery display little changes after the long-term hypoxia, having different functions from the carotid body. The carotid body rudiment is formed in the lateral wall of the third branchial artery. The neural cells immunoreactive for TuJ1, PGP 9.5, and HNK-1, which are continuous with the inferior vagal (nodose) ganglion, first surround and then invade both the carotid body rudiment and the other portions of the third branchial artery, becoming glomus cells.

Responses of the ultimobranchial gland to vitamin D3 treatment in freshwater mud eel, Amphipnous cuchia, kept in different calcium environments.

Freshwater mud eel, Amphipnous cuchia, were injected intraperitoneally daily with 100 ng of vitamin D3/100 g body weight and maintained in media containing either no calcium or different calcium concentrations. The eels were killed after 1, 3, 5, 10 and 15 days following the treatment and their serum calcium levels were measured. The ultimobranchial glands were fixed and processed using the routine paraffin method for histological studies. The results of the present study indicate that vitamin D3 can induce hypercalcaemia in eels kept in different calcium environments. Also, the ultimobranchial glands became hyperactive following vitamin D3 treatment. It is concluded that in mud eels, the gland has a calcium-regulating function.

Direct raises in blood Ca levels by infusing a high-Ca solution into the blood stream accelerate the secretion of calcitonin from the ultimobranchial gland in eels.

In eels, a CaCl(2) solution was infused into the pneumatic duct vein. Plasma Ca levels were significantly increased during 3 hr and were followed by significant raises in plasma calcitonin levels. These results strongly suggest that, in eels, direct raises in blood Ca levels by infusion of a high-Ca solution via blood vessels can accelerate the secretion of calcitonin from the ultimobranchial gland.

Eya1is required for the morphogenesis of mammalian thymus, parathyroid and thyroid

Eyes absent (Eya) genes regulate organogenesis in both vertebrates and invertebrates. Mutations in human EYA1 cause congenital Branchio-Oto-Renal (BOR) syndrome, while targeted inactivation of murine Eya1 impairs early developmental processes in multiple organs, including ear, kidney and skeletal system. We have now examined the role of Eya1 during the morphogenesis of organs derived from the pharyngeal region, including thymus, parathyroid and thyroid. The thymus and parathyroid are derived from 3rd pharyngeal pouches and their development is initiated via inductive interactions between neural crest-derived arch mesenchyme, pouch endoderm, and possibly the surface ectoderm of 3rd pharyngeal clefts. Eya1 is expressed in all three cell types during thymus and parathyroid development from E9.5 and the organ primordia for both of these structures failed to form in Eya1(-/-) embryos. These results indicate that Eya1 is required for the initiation of thymus and parathyroid gland formation. Eya1 is also expressed in the 4th pharyngeal region and ultimobranchial bodies. Eya1(-/-) mice show thyroid hypoplasia, with severe reduction in the number of parafollicular cells and the size of the thyroid lobes and lack of fusion between the ultimobranchial bodies and the thyroid lobe. These data indicate that Eya1 also regulates mature thyroid gland formation. Furthermore, we show that Six1 expression is markedly reduced in the arch mesenchyme, pouch endoderm and surface ectoderm in the pharyngeal region of Eya1(-/-) embryos, indicating that Six1 expression in those structures is Eya1 dependent. In addition, we show that in Eya1(-/-) embryos, the expression of Gcm2 in the 3rd pouch endoderm is undetectable at E10.5, however, the expression of Hox and Pax genes in the pouch endoderm is preserved at E9.5-10.5. Finally, we found that the surface ectoderm of the 3rd and 4th pharyngeal region show increased cell death at E10.5 in Eya1(-/-) embryos. Our results indicate that Eya1 controls critical early inductive events involved in the morphogenesis of thymus, parathyroid and thyroid.

Parathyroid and ultimobranchial glands: Calcium metabolism in birds

Calcium is essential for normal metabolic processes in birds. Several hormonal systems exist to regulate calcium levels. These include calcitonin from the ultimobranchial glands, parathyroid hormone from parathyroid tissue, vitamin D, and the reproductive hormones, especially estrogens. These systems interact closely to keep plasma and tissue calcium levels at a steady state. Calcium disorders may be caused by a number of factors, and can lead to a variety of clinical problems in pet birds. The physiology of calcium metabolism in avian species is discussed along with common disease states arising from derangements in calcium homeostasis.

Calcitonin: characterisation and expression in a teleost fish, Fugu rubripes.

The present report describes the structure and expression of the calcitonin gene in Fugu rubripes. It is composed of 4 exons and 3 introns. Splicing of exons 1, 2 and 3 generates the calcitonin pre-proprotein, while splicing of exons 1, 2 and 4 generates calcitonin gene-related protein (CGRP). Exons 1 and 2 encoding the signal sequence and the N-terminal peptide are common in both the gene products and this gene organisation has been conserved in human, rat, chicken and salmon. The gene environment around calcitonin in Fugu has been poorly conserved when compared with human, apart from a small gene cluster. The calcitonin gene in Fugu has a widespread tissue distribution but it is most highly expressed in the brain. The abundance of gene expression in the ultimobranchial gland and the pituitary indicates that these are important sites of production and that the peptide is probably secreted into the circulation and/or acts as a paracrine or autocrine controlling factor. Whilst the function of calcitonin in fish is still largely unknown, the distribution described here suggests that one of the potential functions may be as a neuropeptide.

Carcinoma misto medular-papilar de tireóide: relato de caso

Casos de carcinomas mistos medular-papilar são raros e não foram classificados pela OMS. Relatamos o caso de uma paciente de 32 anos, com nódulo cervical há 4 anos, sem história familiar de doença nodular tireóidea ou de irradiação prévia. A citologia revelou neoplasia de células de Hürthle e, após tireoidectomia total, o diagnóstico histológico foi de um carcinoma misto: medular-papilar com diferenciação oxifílica, apresentando arranjos papilares com eixos fibro-vasculares contendo predomínio de células de padrão oxifílico, tanto no nódulo como nos gânglios cervicais metastáticos. Notou-se, ainda, a presença de material amorfo de aspecto amilóide que corou positivamente para vermelho Congo, acompanhado de focos de calcificação, tanto no tumor primário quanto nas metástases. A tireoglobulina não foi imunorreativa em células da citologia, porém foi reativa em áreas dos cortes histológicos, tanto no nódulo tireóideo como nas metástases. A imunorreatividade à calcitonina foi predominante nas células da citologia, nos cortes histológicos do tumor primário e das metástases. Levanta-se a hipótese de que tais tumores surjam da transformação neoplásica de um único tipo celular. Recentemente, tem sido sugerido que do último corpo branquial do homem possam surgir ambas as células e que uma célula tronco poderia ser a precursora destas neoplasias.

C-cell and thyroid epithelial tumours and altered follicular development in transgenic mice expressing the long isoform of MEN 2A RET.

Gain-of-function mutations in the gene encoding the receptor tyrosine kinase RET have been identified as the aetiological factor for multiple endocrine neoplasia type 2A (MEN2A). MEN2A is a dominantly-inherited cancer predisposition syndrome characterized by medullary thyroid carcinoma, a tumour of the calcitonin-producing thyroid C-cells. There are three isoforms of RET: RET9, RET43 and RET51, and although in vitro evidence suggests they vary in cellular transformation activities, little is known about their function in tumorigenesis in vivo. To address this, we used RET51 cDNA to construct mice in which the most frequent MEN2A mutation, Cys-634-Arg, was expressed under the control of the human calcitonin promoter (CT-2A mice). These mice developed C-cell tumours resembling human MTC and follicular tumours resembling human papillary thyroid carcinoma (PTC) depending on the founder line examined. One founder line developed compound MTC/PTC at low frequency (8%) and pancreatic cystadenocarcinoma. CT-2A mice also displayed a developmental defect in thyroid follicular structure, in which much of the thyroid was occupied by large irregular cystic follicles thought to be derived from the ultimobranchial body, a developmental precursor of the thyroid gland. The CT-2A mice will provide a suitable model to further study the effects of the MEN 2A RET mutation in vivo.

Neuronal properties in cultured ultimobranchial C cells of chick embryos: process outgrowth and expression of TuJ1 and enkephalin

We have analyzed the neuronal properties in cultured ultimobranchial C cells isolated from embryonic chicks at different developmental stages (12–16 days of incubation) by immunohistochemistry and electron microscopy. The ultimobranchial glands mostly consist of C cell solids. In 13-day-old embryos, many C cells cultured for 7 days on laminin-coated slides extended long neurite-like processes, reaching 300 μm in length. Neuritic outgrowth of C cells was regulated developmentally and virtually unaffected by nerve growth factor (NGF). The cultured C cells expressed intense immunoreactivity for calcitonin and enkephalin. It was also confirmed by confocal laser-scanning microscopy that almost all C cells were intensely immunostained by both the calcitonin antiserum and the monoclonal antibody TuJ1, a neuron-specific marker. Scanning electron microscopy identified the outgrowth of long, branching neuritic processes emerging from C cell soma. The processes had numerous varicosities along their course and ended in growth cones. The C cells with processes were usually monopolar and less frequently bipolar or multipolar. Transmission electron microscopy revealed the presence of membrane-bounded secretory granules in the cultured C cells. The neuritic processes of C cells contained aggregations of microfilaments, intermediate filaments and microtubules arranged in parallel to the long axis. In addition, synaptic-like contacts showing desmosome-like membrane-thickenings and accumulations of small clear vesicles and dense-cored vesicles were formed between the endings of the processes and the surface of C cells. These results indicate that the C cells cultured from early chick embryos (12- and 13-day-old) maintain the neuronal characteristics for long periods in vitro.

A morphological and immunohistochemical study of the ultimobranchial body in the Japanese lizard and the snake.

A morphological and immunohistochemical study of the ultimobranchial body of reptiles Japanese lizard and snake was carried out. The ultimobranchial body of the Japanese lizard was located adjacent to the left arch of the aorta between the trachea and esophagus. It was found as a cluster or group of cells with no capsule. Grimelius' silver impregnation and lead-hematoxylin staining produced positive reactions in some of the clustered cells and follicular cells. The same reaction pattern was observed with anti-calcitonin using the PAP method. The PAP reactions were positive to antiserum against pig calcitonin, but negative to antiserum against synthesized human calcitonin. Furthermore, the PAP reactions were negative to antiserum against tyrosine hydroxylase. The immunofluorescent study of the snake ultimobranchial body revealed that most of the clustered cells and some of the follicular cells were calcitonin-immunoreactive but none was tyrosine hydroxylase-immunoreactive. Certain histological similarities exist between the Japanese lizard ultimobranchial body and snake ultimobranchial body, but the distribution of calcitonin-positive cells were slightly different. In the Japanese lizard, the positive cells were scattered between the foliicles and the number was small. However, most of the cells which formed the cluster in the ultimobranchial body of snake were positive. The findings suggest that the configuration of amino acid in the Japanese lizard calcitonin and snake calcitonin are similar to that of pig calcitonin, and the reptile and the birds is a boundary of the tyrosine hydroxylase existence.

Ultimobranchial Glands in the Teleost (Plecoglossus altivelis ryukyuensis): Special References to Changes of Gland Volume with Maturation

In lower vertebrates, the bone-mineralizing hormone "calcitonin" is secreted from ultimobranchial glands, which assume various shapes, e.g., follicles, cellular masses, or cell strands. Histological observations support the view that in some teleosts, the glands increase in size when females maturate. We determined the exact volume of the gland in juveniles and adult males and females of a teleost, Ryukyuayu (Plecoglossus altivelis ryukyuensis). Furthermore, we examined plasma Ca, Na and K levels. In this species, the gland was fundamentally composed of a single follicle. The gland volume was converted to numerical data under a certain condition. It thus became clear that the value of the follicle wall was significantly increased only in maturing and mature females with high plasma Ca levels and that the value of the lumen did not show any statistically significant changes during growth and maturation.

Expression and development of the proenkephalin mRNA in the C cells of chicken ultimobranchial glands

A large number of enkephalin-immunoreactive cells transiently appear in chick ultimobranchial glands during embryonic development. The expression and development of proenkephalin mRNA were examined in the ultimobranchial glands by in situ hybridization with digoxigenin (DIG)-labeled oligonucleotide probes, in comparison with those of calcitonin mRNA and enkephalin peptide. Proenkephalin mRNA, as well as calcitonin mRNA, appeared in some C cells at embryonal day 14 (E 14), and in many cells at E 16. Subsequently, there is a marked increase in the level of calcitonin mRNA around E 18–19; all C cells exhibited intense reaction for calcitonin mRNA. After hatching, intensity of calcitonin mRNA expression was more and more increased. Northern blot analysis with the calcitonin probe also indicated that calcitonin synthesis of the C cells progressively increased with developmental gradient, and reached to the adult level at 1 month after hatching. On the other hand, intensity of hybridization signal of proenkephalin mRNA was maintained moderately during development. In contrast to enkephalin immunoreactivity, which is markedly decreased after hatching, proenkephalin mRNA expression was consistently detected in many C cells of 1- and 2-month-old chickens. Reverse transcription–polymerase chain reaction (RT–PCR) analysis confirmed that proenkephalin mRNA was obtained in the ultimobranchial glands of not only embryos but also 1-day- and 1-month-old chickens. Furthermore, Northern blot analysis demonstrated that a single band for proenkephalin mRNA was obtained in the poly (A) +RNA isolated from the ultimobranchial gland of 1-day-old chicks. Thus, the present study evidences that proenkephalin mRNA is synthesized in almost all C cells of chicken ultimobranchial glands throughout life. Enkephalin may be essential for C cell function.

Anuran Calcitonins are Diverse in Lower Vertebrates

We investigated the nucleotide sequences of cDNA fragments coding calcitonin from ultimobranchial glands in 2 species of urodelans (1 salamander and 1 newt) and 4 species of anurans (1 toad and 3 frogs) by the reverse transcription polymerase chain reaction (RT-PCR) method and rapid amplification of cDNA ends (RACE)-PCR method. The salamander and newt calcitonins were each 97% and 94% similar to the lungfish and caiman calcitonins that we have already reported, in the amino acid sequences. However, anuran calcitonins were not only dissimilar (63-81%) to the lungfish and caiman calcitonins but also diversified (59-91%) even among anurans. The sequence identity of toad calcitonin was always low (59-66%) among anurans.

Posterior Mediastinal Goiters: Literature Review and Report of Three Cases

Posterior mediastinal goiters are very rare and occur mostly due to descent of a posterolaterally enlarging inferior pole of the thyroid gland, or very infrequently to failure of fusion of the ultimobranchial bodies with the isthmus in the 7th embryonic week. Most patients present with a cervical mass and symptoms due to compression or distortion of the trachea, esophagus, or superior vena cava. The diagnosis is established by chest skiagram, computed tomography scan, and barium esophagogram. Progressive enlargement, risk of sudden hemorrhage within the gland causing respiratory impairment, and the possibility of associated malignancy, make excision of the goiter mandatory. A combined cervicothoracic approach is the procedure of choice as it provides easy access and visualization, better control of blood vessels, and avoids the risk of perioperative tumor seeding. We present our experience of 3 such cases successfully managed at our institute. All 3 patients presented with a cervical mass and symptoms of posterior mediastinal compression; one had thyrotoxicosis. A combined cervicothoracic approach was used for surgical excision with excellent results.

Topography and immunohistochemistry of the ultimobranchial glands in some subtropical fishes

Topography and immunohistochemistry of the ultimobranchial glands in some subtropical fishes

Fish Calcitonin Genes: Primitive Bony Fish Genes Have Been Conserved in Some Lower Vertebrates

Using the polymerase chain reaction method, we amplified calcitonin genes from the cDNA of the ultimobranchial glands or from the genomic DNA of the blood cells and liver of various fishes. The fishes examined were primitive bony fishes (lungfish, polypterus, sturgeon, and gar), 16 species of teleosts, and cartilaginous fishes (stingray). Sequenced calcitonin genes were compared among fishes and with those of reptiles and chickens. The similarity of the calcitonin genes was the highest between the reptile and chicken groups and the primitive bony fishes (sequence similarity of the nucleotides 81–90%). The values between teleosts and the primitive bony fishes were 70–81%, with the exception of eels. Eel calcitonin genes were very similar to those of primitive bony fishes (83–88%). Stingray calcitonin genes were relatively more similar to those of the primitive bony fishes (74–78%) than to teleosts (63–73%). In goldfish and sardine, two types of calcitonin genes were found. We concluded that the genes of primitive bony fish are placed at a fundamental position in this hormone, at least among these vertebrates.

Parathyroids and ultimobranchial bodies in monotremes

Parathyroids and ultimobranchial bodies in monotremes