In vivo shedding of apical plasma membrane in the thyroid follicle cells of the mouse

Summary Clusters of luminal dense bodies, limited by a triple-layered membrane, were found in all follicle lumina in thyroid glands of mice. After thyroxine treatment the number of luminal dense bodies increased, especially in the periphery of the lumen, where the intraluminal bodies often displayed a striking resemblance to microvilli. In hyperplastic goiters, obtained by feeding mice with propylthiouracil, luminal dense bodies were replaced by intraluminal vesicles. During goiter involution the vesicles were gradually replaced by luminal dense bodies; the presence of intermediate forms suggests that vesicles and dense bodies are basically the same formations. Luminal dense bodies were observed in colloid droplets indicating their removal by endocytosis. As demonstrated by electron-microscopic cytochemistry, luminal dense bodies contain a membranebound peroxidase, and electron-microscopic autoradiography after administration of ¹²⁵I indicate that they possess an iodinating capacity.Our observations on mouse thyroid glands suggest that the luminal dense bodies, which appear as vesicles in hyperplastic glands, are formed by shedding of the apical plasma membrane of the follicle cell. The shedding process might be of importance for the turnover of plasma-membrane material.This study was supported by Grant No. 12X-537 from the Swedish Medical Research Council.     

X-ray microanalysis on the thyroid follicle of the hagfish, Eptatretus burgeri and lamprey, Lampetra japonica.

Intracellular dense bodies, cytoplasmic matrices, and luminal colloids in the thyroid follicle of cyclostomes, hagfish and lamprey were examined to identify the distribution of iodine using an energy dispersive X-ray microanalyzer attached to a scanning transmission electron microscope. High level of iodine was detected only in the large dense body of the hagfishes, while it was too slight in quantity to detect by this method in the cytoplasmic matrix as well as in the luminal colloid. In the adult lamprey thyroid, an appreciable amount of iodine was detected in a few large dense bodies. In mice and rats, it is very hard to detect iodine in the luminal colloid, intracellular colloid droplet, and in the lysosomal dense granule by this method, though these structures have been well known to contain a fairly large amount of iodine which is combined with thyroglobulin. These facts mean that intracellular dense bodies in the thyroid follicular epithelium of the cytoclostome, especially of the hagfish have an extremely larger amount of iodine. These bodies are considered to be secondary lysosomes or residual bodies containing reabsorbed colloid materials which are highly condensed.     

Evaluation of the utility of air-dried whole mounts for quantitative electron microprobe studies of platelet dense bodies.

A variety of electronmicroscope techniques have been used to examine how the air-drying process may affect the dense bodies in whole mounts of platelets. (a) Selected-area-diffraction and electron microprobe studies suggest that the air-drying process can result in the formation of crystalline precipitates of sodium chloride on grid films and platelets. However, no crystals were detected in the calcium-and-phosphorus-containing matrix of dense bodies. (b) Tilting studies show that dense bodies in human platelets are spherical or ellipsoidal in shape. Dense bodies in rabbit platelets, in contrast, appear flattened in a horizontal plane. (c) Human-platelet dense bodies probed with a small (20 nm diameter) spot vary widely in their peak/background ratios for calcium and phosphorus-a finding that suggests that the two elements may not be evenly distributed throughout the dense-body matrix. Nevertheless, when dense bodies are probed with a larger (200 nm diameter) spot, they do not appear to differ appreciably among themselves in their calcium or phosphorus content. The data suggest that with human platelets, air drying may be a preparative procedure which permits comparison by microprobe techniques of dense-body matrix content in platelet populations.     

Fine structural aspects of the black thyroid induced by minocycline, and the effects of a low iodine diet, propylthiouracil, thyroxine tablet and TSH, on the black discoloration of the rat thyroid

Fine structural aspects of the effect of minocycline, an antibiotic of the tetracycline group, on the rat thyroid were studied. In all the rats administered minocycline (100 mg/kg/day) for 21 days, diffuse black discoloration of the thyroid gland occurred. However, when the rats were fed on a low iodine diet, given propylthiouracil (PTU) or thyroxine tablet with minocycline the black pigmentation of the thyroid gland did not take place. On the other hand, black discoloration of the thyroid was accelerated in the rats administered TSH and minocycline simultaneously. Ultrastructurally, numerous dense bodies containing highly electron-dense deposits were seen in the supranuclear region of the follicular epithelial cells of the black thyroid. These dense bodies, which showed positive acid phosphatase activity, are considered to be lysosomes containing minocycline or its derivatives. It is speculated that minocycline is taken up into follicular epithelial cells with iodine, and that the black discoloration of the thyroid gland is intimately related to iodine metabolism.     

THE ORIGIN OF PROTEIN AND FATTY YOLK IN RANA PIPIENS : II. Electron Microscopical and Cytochemical Observations of Young and Mature Oocytes

Electron microscope studies of young oocytes have demonstrated that the plate-like, hexagonally shaped yolk bodies previously observed in living cells are wholly within the substance of oocyte mitochondria and that they remain within these mitochondria while increasing in size. These bodies possess a crystalline structure consisting of what appear to be lines, with a spacing of 70 to 85 A, and appear very dense in the electron microscope. After formalin fixation such bodies give an intense positive test for protein, and when viewed in the electron microscope are only slightly less dense than after OsO₄ fixation. Evidence is presented for the origin of these crystals within a single crista. The clusters of yolk globules previously studied in living cells are seen to consist of several types of bodies, but an irregular dense droplet predominates. This dense material is apparently secreted by small spherical bodies which, the evidence suggests, originate from the breaking up of filamentous mitochondria and which possess an outer double membrane and sometimes internal cristalike membranes. When thin sections of young oocytes are immersed in xylol the dense globules of the clusters are dissolved, but the hexagonal bodies are unaffected, indicating that the globules are of a predominantly fatty nature, while the hexagonal bodies are of a predominantly protein nature. Examination of mature or almost mature oocytes has revealed that the main body of the yolk platelets is crystalline in nature and is surrounded by a thick matrix which, in light microscope study, masks the fact that the face view of the main body of the platelets is often hexagonal. The spacing within the main body is found to be 70 to 85 A. The crystal laminae of this material can be resolved quite clearly into rows of particles. Dense globules of varying sizes are found in the cytoplasm between the platelets. When thin sections of these OsO₄-fixed oocytes are immersed in xylol, the material of the globules is extracted and the crystalline material of the platelets remains unaffected, indicating the fatty nature of the globules and the protein nature of the platelets. The platelets of the mature egg resemble the hexagon bodies, previously described in young oocytes, in their protein nature, their crystalline spacing, and their hexagonal outline. This is given as strong evidence for the origin of the mature platelets by the growth of the intramitochondrial hexagon bodies. The biochemical implications of this study are discussed.     

Lithium administration affects gene expression of thyroid hormone receptors in rat brain.

Even though lithium has received wide attention in the treatment of manic depressive illness, the mechanisms underlying its mood stabilizing effects are not understood. Lithium is known to interact with the thyroid axis and causes hypothyroidism in a subgroup of patients, which compromises its mood stabilizing effects. Since lithium was recently reported to alter thyroid hormone metabolism in the rat brain, the present study investigated whether these effects were mediated through regulation of thyroid hormone receptor (THR) gene expression. Adult male euthyroid rats were either given a diet containing 0.25% lithium or one without lithium for 14 days. Rats were sacrificed in the evening and RNA was isolated from different brain regions to quantitate the isoform specific mRNAs of THRs. Following 14 days of lithium treatment, THR alpha1 mRNA levels were increased in the cortex and decreased in hypothalamus; THR alpha2 mRNA levels were increased in the cortex and THR beta mRNA levels were decreased in the hypothalamus. No significant difference in the expression of these THR isoforms was observed in the hippocampus or cerebellum. Thus, chronic lithium treatment appeared to regulate THR gene expression in a subtype and region specific manner in the rat brain. It remains to be determined whether the observed effects of lithium on THR gene expression are related to its therapeutic efficacy in the treatment of bipolar disorder.     

Fine structural aspects on the fate of rat black thyroids induced by minocycline

The fate of the black thyroid induced by minocycline treatment (100 mg/kg daily for 21 days) in the rat was studied by light and electron microscopy after 6 months. The black discoloration of the thyroid gland remained and numerous dense bodies containing highly electron-dense deposits were seen in most of the follicular epithelial cells. It appears that the turnover rate of follicular epithelial cells is very low and the electron-dense deposits are largely retained, though debris derived from degenerate follicular epithelial cells containing the dense deposits may be phagocytosed by macrophage-like cells in the interfollicular connective tissue. Brown-black granules are also found in the extremely attenuated follicular epithelial cells of cold follicles.     

Immunocytochemical localization of cathepsins B, H, L, and T4 in follicular cells of rat thyroid gland

To localize cathepsins B, H, and L in follicular cells of rat thyroid gland, we applied immunocytochemistry to the thyroid tissue using their respective monospecific antibodies. On serial semi-thin sections, cathepsins B, H, and L were localized in granules of various sizes located throughout the cytoplasm, whereas T4 was detected in larger granules located in the apical and supranuclear regions. By electron microscopy, cathepsins B, H, and L were localized in large less-dense granules (so-called colloid droplets) and in dense bodies of various sizes, whereas T4 was localized more intensely in large less-dense granules than in smaller dense bodies. By double immunostaining using an immunogold method, cathepsins H and B or L were co-localized in the same cytoplasmic granules. Moreover, immunoblotting demonstrated that proteins similar to cathepsins B, H, and L in the liver are present in the thyroid gland. These results suggest that cathepsins B, H, and L participate not only in degradation of thyroglobulin but in maturation of thyroid hormones, although it remains unknown whether all of them participate in the maturation process.     

Fine needle aspiration cytology in a case of diffuse sclerosing carcinoma of the thyroid

The fine needle aspiration (FNA) cytologic findings in a rare case of bilateral diffuse sclerosing carcinoma of the thyroid are described. Along with a generally papillary architecture, the FNA biopsy smears showed a number of features not usually seen in thyroid carcinomas: psammoma bodies, an abundance of squamous metaplasia and a dense inflammatory component. The FNA diagnosis was multifocal papillary carcinoma with squamous metaplasia; histopathologic examination made the final diagnosis.     

Ultrastructural pathology of the thyroid folliculo-papillary and follicular carcinoma

Twenty cases of thyroid carcinoma (10 follicular and 10 folliculo-papillary) were ultrastructurally studied. In the follicular carcinoma the most striking features were: microfollicular cavities with microvilli from the apical surface of the tumorous cells, intracellular microlumens, swollen mitochondria sometimes containing electrondense bodies and tightly packed filaments. In the solid sheaths light and dark cells were present. Golgi complexes were disposed in small dense cristae. The nuclei were large, round, oval or with a folded appearance. In the folliculo-papillary carcinoma were found nuclei with an irregular shape containing stage I and stage II inclusions, dilated endoplasmic sacks, closely packed, sometimes dystrophic mitochondria, dense bodies or tightly packed parallel filaments and numerous phagolysosomes. The peroxidase activity wa present as black precipitates in the nuclear envelope or around colloid droplets. The acid phosphatase activity was found as unhomogeneous precipitates inside the lysosomes. From this study it could be concluded that the follicular and folliculo-papillary carcinomas have some common ultrastructural features; the ultrastructural and cytoenzymological patterns suggest marked alteration of the synthesis, storage and secretion of thyroid hormones.     

Ultrastructural studies of normal and alloxan-treated islet cells of the pancreas of the lizard, Eumeces fasciatus

Alpha and beta cells can be distinguished by differences in mitochondrial and secretion granule structure. Many mitochondria of alpha cells possess “tubular” or prismatic cristae oriented longitudinally and having triangular profiles in cross-section. The matrix is particulate, with the roughly spherical particles measuring about 100 A in diameter. Evidence is presented indicating that alpha and beta granules are sequestrated in association with Golgi elements. Fully-condensed beta granules, assumed to be insulin, appear homogeneously dense and crystalline. Recurrent profiles of crystalline beta granules suggest that they possess an octahedral configuration. Alpha cell granules also appear homogeneously dense but have round profiles. Many acinar cell nuclei in both normal and alloxan-treated pancreata display masses of moderately dense fibrils oriented roughly parallel to each other. These fibrils are about 200 A across and their terminal portions are rebranched and often appear to be continuous with the finely granular or filamentous component of the nucleoplasm. Not infrequently the fibrils show evidence of periodicity. Alloxan has a specific destructive effect on beta cells. An initial effect seems to be the disruption and coalescence of the bounding membranes of beta granules. Lysosome-like bodies are often seen in beta cell cytoplasm, which ultimately becomes degranulated and necrotic following prolonged administration of large doses of alloxan.     

Crystalline inclusions in hepatocytes and associated interhepatocytic macrophages from female Ohrid trout (Salmo letnica Kar.)

When characterizing the liver ultrastructure in Ohrid trout, we noticed that cells often displayed unusual cytoplasmic crystalline inclusions. Their morphology varied much, concerning the size, shape and electron density, showing aspects never reported in fish liver. In hepatocytes, the inclusions existed in close topographical relationship with dense bodies (often within them). Such “crystals” inclusions also existed in macrophages and in biliary passages lumina. Data revealed seasonal/breeding-related changes of the dense bodies and crystalline inclusions within hepatocytes; decreasing from the earliest to the more advanced ovary maturation stages. Additionally, based on the negative correlations between the amounts of dense bodies and of “crystals” and the ovary somatic ratio, we suggested there might be connections between the sex steroids status and the inclusions formation. A positive correlation between the quantities of “crystals” and of dense bodies further suggested that the inclusions might derive from normal processes, e.g., the females’ cyclic liver changes, that involves dense bodies’ turnover. However, a toxicological aetiology cannot be excluded. Additionally, multiple mechanisms can contribute to the “crystals” genesis. Facing literature reports and because the inclusions content seemed washed out during tissue processing, they are likely lipid in nature, but their exact composition and genesis require further analyses.     

Occurrence of Goitre during Lithium Treatment

Of 330 patients given lithium for recurrent manic-depressive disorder 12 developed goitre after treatment periods of five months to two years. All the patients remained clinically euthyroid. Pressure symptoms necessitated subtotal thyroidectomy in two patients. In 9 out of 10 patients with goitre, and in two out of seven without goitre study with radioactive iodine showed abnormal findings in iodine metabolism. Discontinuance of lithium led to disappearance of goitres, while thyroid metabolism returned to normal. Thyroxine or desiccated thyroid produced shrinkage of the gland in spite of continued lithium medication.     

Electron microscopical studies on the thyroid of a cyclostome,Lampetra japonica,during its upstream migration

Summary Electron microscopical studies were made of the thyroid gland of an adult lamprey, Lampetra japonica, in the upstream migration period.The thyroid consists of many usual follicles containing the colloid in their lumina, and a large parafollicle without colloid. The paper concerns only the usual follicle.The follicle cells found in the usual follicle wall are classified into three types; 1. a non-ciliated taller cell, 2. a ciliated taller one, and 3. a non-ciliated cuboidal one. From their cytoplasmic fine structure, it is considered that all these cells are essentially identical and differences among them are due to their functional state.All these type cells are characterized by irregularly developed interdigitations and aggregates of tonofilaments throughout the cytoplasm, especially in the perinuclear region. Although the rough-surfaced endoplasmic reticulum and the Golgi apparatus are fairly well developed in the first and second type cells, the cisternae are not so large-vacuolated but flattened, and the cytoplasm is more compact as compared with that of the higher vertebrate. In the third type cell, the cytomembranes are poorly developed.Large dense inclusion-bodies consisting of heterogeneously dense materials, of lamellar structures, and of less dense vacuoles, which are found often in taller follicle cells, are also characteristic for the lamprey thyroid. The body which might be intimately related to the Golgi apparatus is considered to be a kind of lysosomes and it perhaps corresponds to the yellow pigment observed by light microscopy.In the apical part of the cytoplasm in taller cells, there are three kinds of granules or vesicles; numerous small vesicles considered to be derived from the Golgi apparatus, a few small dense granules which seem to originate from the Golgi region, and a few large less-dense granules.In the third type cell, the cytomembranes are not so well developed as those of the first and second type cells. The large heterogeneously dense bodies and the cytoplasmic granules are very few in number.Around the follicle of the lamprey thyroid, there are a dense basement membrane and a relatively compact connective tissue with few blood capillaries. Characteristic fat cells are found in the connective tissue.     

Biokinetics of iodine-131 in rat thyroid following lead and lithium supplementation

The impact of lead as an environmental pollutant on the I-131 uptake and retention in rat thyroid was assayed alone and in combination with lithium treatment. Lead treatment significantly stimulated the 2- and 24-h uptake of I-131 in the thyroid, and the 24-h uptake showed the maximum stimulation after 3 mo of lead treatment. On the contrary, lithium supplementation reduced the uptake significantly and the maximum decrease was noticed after 2 mo of lithium administration. Further, simultaneous lead and lithium treatment resulted in more pronounced increase in the uptake of I-131 by the thyroid, which was maximum after 3 mo of combined treatment. The thyroidal biological half-life of I-131 (T _biol) was found to be increased significantly following lead and lithium treatments when given separately. Interestingly, combined lead and lithium treatment given up to 2 mo further prolonged theT _biol of I-131, thus reflecting its increased retention.     

Ultrastructure of crystalline inclusions in the thylakoids of dodder<Emphasis Type="Italic">(cuscuta japonica)</Emphasis> plastids

The plastids from seedlings of the parasitic angiospermCuscuta japonica were ultrastructurally investigated. In shoot subapical cells from 3-d-old seedlings grown in the dark, the etioplasts contained prolamellar bodies and amorphous and dense inclusions. In the shoot subapical cells obtained from 6-d-old seedlings grown under light conditions for the last 3 d, the underdeveloped chloroplasts contained phytoferritin within the stroma as well as amorphous and dense inclusions that were limited by the thylakoid membranes. In the developing chloroplasts, electron-dense materials were detected within the transversely sectioned thylakoid lumens. This dense material presented two different images, depending upon the sectional plane. When transversely prepared, the materials appeared as somewhat thick, linear structures, whereas longitudinally sectioned thylakoids revealed very large crystalline inclusions. In the developed chloroplasts, the amounts of electron-dense material or crystalline inclusions were remarkably reduced in the thylakoid lumens, which were electron-translucent. Far fewer crystalline inclusions were observed in the developed chloroplasts of seedlings than in the developing chloroplasts. These results suggest that the crystalline inclusions may be temporarily reserved within the thylakoid lumens of chloroplasts in the Gjaponica seedlings.     

The Intracellular Localization of Hormonal Activity in Transplantable Thyrotropin-Secreting Pituitary Tumors in Mice

Mouse pituitary tumors secreting almost exclusively thyroid stimulating hormone have been characterized electron microscopically. Tumors of known thyrotropin content were separated into nuclear, mitochondrial, microsomal, and soluble fractions by differential centrifugation. The hormonal activity of these fractions was correlated with that of the total homogenates and with their nitrogen and phosphorus content. Essentially all the thyrotropin of the homogenate was recovered in a particulate fraction sedimenting between 20,000 and 40,000 g. This fraction contained the RNA granules and membranous components typical of microsomal pellets, but also showed the presence of small dense bodies surrounded by smooth membranes. These bodies were also visible within the endoplasmic reticulum of intact cells, and it is postulated that these bodies may represent the sites of intracellular elaboration and/or storage of TSH. Thyrotropin is tightly associated with microsomal particles but can be brought into solution by treatment with alkaline media, deoxycholate, and certain organic solvents.     

DISTRIBUTION OF PHOSPHATASES IN THE GOLGI REGION AND ASSOCIATED STRUCTURES OF THE THORACIC GANGLIONIC NEURONS IN THE GRASSHOPPER, MELANOPLUS DIFFERENTIALIS

The neuronal perikarya of the grasshopper contain sudanophilic lipochondria which exhibit an affinity for vital dyes. These lipochondria are membrane-delimited and display acid phosphatase activity; hence they correspond to lysosomes. Unlike those of most vertebrates, these lysosomes also hydrolyze thiamine pyrophosphate and adenosine triphosphate. Like vertebrate lysosomal "dense bodies," they are electron-opaque and contain granular, vesicular, or lamellar material. Along with several types of smaller dense bodies, they are found in close spatial association with the Golgi apparatus. The Golgi complexes are frequently arranged in concentric configurations within which these dense bodies lie. Some of the smaller dense bodies often lie close to or in association with the periphery of dense multivesicular bodies. Further, bodies occur that display gradations in structure between these multivesicular bodies and the dense lysosomes. Acid phosphatase activity is present in the small as well as the larger dense bodies, in the multivesicular bodies, and in some of the Golgi saccules, associated vesicles, and fenestrated membranes; thiamine pyrophosphatase is found in both the dense bodies and parts of the Golgi complex. The close spatial association of these organelles, together with their enzymatic similarities, suggests the existence of a functional or developmental relationship between them.     

Effect of short-term and long-term lithium treatment on uptake and retention of iodine-131 in rat thyroid

No significant change occurred in the uptake by the thyroid of male Wistar rats of a standard dose of carrier-free 131I administered intraperitoneally and its retention by the thyroid, as measured by biological and effective half-life, after feeding these rats a powdered pelleted diet containing lithium carbonate (1.1 g per kg of diet) for 7 days. However, continuing this diet for 10 days inhibited thyroid uptake and increased the retention of 131I. Uptake remained suppressed for up to 4 months after lithium treatment and continuing this treatment for 6 months did not result in any significant change in 131I uptake by the thyroid. Lithium treatment for 10 days increased the biological and effective half-life of 131I in the thyroid and this increase continued for the 6 months treatment period. The dose of 131I delivered to the thyroid was significantly lower after 10 days and 1 month of lithium treatment but there was no change in this dose after 2 and 4 months of treatment. However, there was a significant increase after 6 months.     

Intracytoplasmic lumina in medullary carcinoma of the thyroid gland. Report of a case with cytologic and immunocytochemical features

BACKGROUND: Intracytoplasmic lumina (ICL) have been observed frequently in breast carcinoma cells. However, they are extremely rare in thyroid gland tumors. We encountered a medullary carcinoma of the thyroid (MCT) with ICL and present a case with cytologic, immunocytochemical and ultrastructural features. CASE: A 15-year-old female was admitted with a left thyroid mass. Ultrasound examination revealed a well-defined tumor in the left lobe of the thyroid. Fine needle aspiration cytology showed mainly dispersed spindle cells with oval nuclei and some polymorphic or triangular tumor cells. The tumor cells containing ICL were noted at high magnification. The ICL contained sparse microvilli and abundant granular material with dense, round bodies on ultrastructural sections. Immunocytochemically, these tumor cells were positive for calcitonin and carcinoembryonic antigen (CEA). Moreover, CEA was recognized in the ICL with immunocytochemical staining. All tumor cells were negative for thyroglobulin. Pathologic examination confirmed the diagnosis of medullary carcinoma of the thyroid gland. CONCLUSION: MCT can include ICL with granular material containing CEA.