Follicular Lumen Research Articles

Pinopsin and photoreception in the pineal organ of the domestic turkey during post-hatching development

The avian pineal organ is photosensitive because of the presence of photopigments, of which pinopsin seems to be one of the most important. This organ is subject to far-reaching changes during post-hatching development, but evidence regarding pinopsin presence and direct photoreception during this time is lacking. This study was carried out to demonstrate the following: 1) the structures showing immunoreactivity to pinopsin in the turkey pineal organ, 2) the changes of these structures during development, 3) the pinopsin localization in pinealocytes in monolayer cultures, and 4) the role of direct photoreception in the regulation of melatonin secretion in pineal organs in adult turkeys. Pinopsin immunoreactivity was localized in the apical extensions of columnar cells limiting the follicular lumen, in fiber-like structures located between columnar cells in the inner part of follicle wall, in string-shapes or small spherical structures distributed in the outer part of follicle wall and in amorphous material inside the follicle lumen. In young birds, immunoreactivity was also sporadically noted in cell bodies of rudimentary receptor pinealocytes. The distribution of pinopsin showed prominent age-dependent changes, including a subsequent increase in pinopsin-positive structures in the outer part of the follicle wall and a prominent reduction in the number and size of positive apical extensions in 40- and 56-week-old turkeys. These data demonstrate that the role of secretory pinealocytes in pineal photoreception increases with age. In monolayer cultures, all pinealocytes showed strong reactions in club- or bulbous-shaped prolongations. The pineal organs of adult birds were less sensitive to light exposition at night than those of young turkeys, which points to differences in light sensitivity between rudimentary receptor and secretory pinealocytes. However, direct photoreception could play an important role in the regulation of melatonin secretion in adult turkeys.

Primary Cilium in the Human Thyrocyte: Changes in Frequency and Length in Relation to the Functional Pathology of the Thyroid Gland.

Primary cilia (PC) are conserved structures in the adult thyroid gland of different mammals. It was recently described that in humans, PC are usually present as a single copy per follicular cell emerging from the follicular cell apex into the follicular lumen. To understand the role developed by PC in thyroid hormonogenesis better, their changes in different human functional thyroid diseases (diffuse toxic hyperplasia/Graves' disease [GD] and nodular hyperplasia [NH]/nodular goiter), in comparison to normal thyroid tissue, were investigated using immunofluorescence, morphometry, and electron microscopy analyses. Significantly decreased ciliary frequencies were found in both NH (51.16 ± 11.69%) and GD (44.43 ± 23.70%) compared to normal thyroid tissue (76.09 ± 7.31%). Similarly, PC lengths were also significantly decreased in both NH (2.02 ± 0.35 μm) and GD (2.4 ± 0.48 μm) compared to normal glands (3.93 ± 0.90 μm). Moreover, in GD patients, hyperactive-follicle foci always showed diminished ciliary frequency and length compared to any other thyroid follicle pattern, independent of their thyroid status. Finally, in GD, the percentage of thyrocytes exhibiting PC in the "normal-appearance areas" was significantly lower in correspondence with the subsistence of signs of thyroid biosynthetic hyperactivity after long-term antithyroid drug treatment. The results suggest a direct relationship between ciliogenesis and both follicle activity and tissue heterogeneity in the functional pathology of the thyroid gland.

The role of thyroglobulin in thyroid hormonogenesis.

In humans, the thyroid hormones T3 and T4 are synthesized in the thyroid gland in aprocess that crucially involves the iodoglycoprotein thyroglobulin. The overall structure of thyroglobulin is conserved in all vertebrates. Upon thyroglobulin delivery from thyrocytes to the follicular lumen of the thyroid gland via the secretory pathway, multiple tyrosine residues can become iodinated to form mono-iodotyrosine (MIT) and/or di-iodotyrosine (DIT); however, selective tyrosine residues lead to preferential formation of T4 and T3 at distinct sites. T4 formation involves oxidative coupling between two DIT side chains, and de novo T3 formation involves coupling between an MIT donor and a DIT acceptor. Thyroid hormone synthesis is stimulated by TSH activating its receptor (TSHR), which upregulates the activity of many thyroid gene products involved in hormonogenesis. Additionally, TSH regulates post-translational changes in thyroglobulin that selectively enhance its capacity for T3 formation - this process is important in iodide deficiency and in Graves disease. 167 different mutations, many of which are newly discovered, are now known to exist in TG (encoding human thyroglobulin) that can lead to defective thyroid hormone synthesis, resulting in congenital hypothyroidism.

Expression, Localization, and Regulation of the Sodium Bicarbonate Cotransporter NBCe1 in the Thyroid.

The intrafollicular space of thyroid follicles is the storage compartment for thyroid hormones. Its pH has been established at around 7.6 at least after thyrotropin (TSH) stimulation. This alkaline intrafollicular pH is thought to be critical for iodide coupling to thyroglobulin and internalization of iodinated thyroglobulin. At least in mice, this alkalinization requires the expression of pendrin (Slc26a4) within the apical membrane, and a lack of pendrin results in acidic follicular lumen pH. Yet, the mechanism importing HCO3- into the cytoplasm is unknown. This study investigated whether the rather ubiquitous sodium bicarbonate cotransporter NBCe1 (SLC4A4) might play this role. It also examined which variant was expressed and where it was localized in both rat and human thyroid tissue. Lastly, the dependence of its expression on TSH was studied. Reverse transcription polymerase chain reaction, immunofluorescence, and Western blotting were used to test whether TSH stimulated NBCe1 protein expression in vivo. Subcellular localization of NBCe1 was performed using immunofluorescence in both rat and human thyroid. Cultured thyroid cells were also used to attempt to define how TSH affects NBCe1 expression. Only transcripts of the NBCe1-B variant were detected in both rat and human thyroid. Of interest, NBCe1-C was not detected in human tissues, not even in the brain. On immunofluorescence microscopy, the immunostaining of NBCe1 mainly appeared in the basolateral membrane upon stimulation with TSH. This TSH induction of basolateral membrane expression of NBCe1 protein was confirmed in vivo in rat thyroid and in vitro on human thyroid slices. This study demonstrates the expression of the sodium bicarbonate cotransporter NBCe1-B in rat and human thyroid. Additionally, the data suggest that TSH blocks the degradation of NBCe1 protein by trafficking it to the basolateral membrane. Hence, TSH increases NBCe1 half-life without increasing its synthesis.

Loss of Primary Cilia Results in the Development of Cancer in the Murine Thyroid Gland.

Communications at the interface between the apical membrane of follicular cells and the follicular lumen are critical for the homeostasis of thyroid gland. Primary cilia at the apical membrane of thyroid follicular cells may sense follicular luminal environment and regulate follicular homeostasis, although their role in vivo remains to be determined. Here, mice devoid of primary cilia were generated by thyroid follicular epithelial cell-specific deletion of the gene encoding intraflagellar transport protein 88 (Ift88 ). Thyroid follicular cell-specific Ift88-deficient mice showed normal folliculogenesis and hormonogenesis; however, those older than 7 weeks showed irregularly dilated and destroyed follicles in the thyroid gland. With increasing age, follicular cells with malignant properties showing the characteristic nuclear features of human thyroid carcinomas formed papillary and solid proliferative nodules from degenerated thyroid follicles. Furthermore, malignant tumor cells manifested as tumor emboli in thyroid vessels. These findings suggest that loss-of-function of Ift88/primary cilia results in malignant transformation from degenerated thyroid follicles.

DUOX Defects and Their Roles in Congenital Hypothyroidism.

Extracellular hydrogen peroxide is required for thyroperoxidase-mediated thyroid hormone synthesis in the follicular lumen of the thyroid gland. Among the NADPH oxidases, dual oxidases, DUOX1 and DUOX2, constitute a distinct subfamily initially identified as thyroid oxidases, based on their level of expression in the thyroid. Despite their high sequence similarity, the two isoforms present distinct regulations, tissue expression, and catalytic functions. Inactivating mutations in many of the genes involved in thyroid hormone synthesis cause thyroid dyshormonogenesis associated with iodide organification defect. This chapter provides an overview of the genetic alterations in DUOX2 and its maturation factor, DUOXA2, causing inherited severe hypothyroidism that clearly demonstrate the physiological implication of this oxidase in thyroid hormonogenesis. Mutations in the DUOX2 gene have been described in permanent but also in transient forms of congenital hypothyroidism. Moreover, accumulating evidence demonstrates that the high phenotypic variability associated with altered DUOX2 function is not directly related to the number of inactivated DUOX2 alleles, suggesting the existence of other pathophysiological factors. The presence of two DUOX isoforms and their corresponding maturation factors in the same organ could certainly constitute an efficient redundant mechanism to maintain sufficient H2O2 supply for iodide organification. Many of the reported DUOX2 missense variants have not been functionally characterized, their clinical impact in the observed phenotype remaining unresolved, especially in mild transient congenital hypothyroidism. DUOX2 function should be carefully evaluated using an in vitro assay wherein (1) DUOXA2 is co-expressed, (2) H2O2 production is activated, (3) and DUOX2 membrane expression is precisely analyzed.

Cytotoxic effects of Euterpe oleraceae fruit oil (açaí) in rat liver and thyroid tissues

Euterpe oleracea Mart., Arecaceae, fruit (açaí) presents considerable potential for the development of new medicines due to its phytochemical composition and antioxidant activity. More recently, special attention has been given to the pharmacological potential of the fruit's oil. This study analysed the histological and histochemical effects of different dosages of açaí oil on rat's liver and thyroid cells, in order to evaluate its cytotoxic potential after administration for consecutive days. Male Wistar rats were treated with the açaí oil by gavage at doses of 30, 100 and 300mg/kg, for 14 days, within a 24h interval. Liver and thyroid fragments were collected for histology (hematoxylin and eosin) and histochemistry analysis (blue of Nilo (lipids), Baker (lipids), bromophenol blue (protein), PAS (polysaccharides)). The results showed that animals exposed to açaí oil presented alterations in the liver cells, where the integrity of the liver tissue was increasingly lost as the açaí oil doses increased. Nuclear pyknosis was observed in several hepatocytes, evidencing the occurrence of cell death. Alteration in the amount of lipids, polysaccharides, vacuoles in the cytoplasm, and proliferation of Kupffer cells were observed in histochemical analyzes. As for the thyroid of the treated rats, alterations were observed in the size of the follicular lumen and also in the connective tissue found between the follicles. Under the experimental conditions employed in the present study, the cytotoxicity observed in this work is worrying, specially considering the liver, when frequent or continuous damage could lead pathological disorders in this organ.

Glycosylation in the Thyroid Gland: Vital Aspects of Glycoprotein Function in Thyrocyte Physiology and Thyroid Disorders.

The key proteins responsible for hormone synthesis in the thyroid are glycosylated. Oligosaccharides strongly affect the function of glycosylated proteins. Both thyroid-stimulating hormone (TSH) secreted by the pituitary gland and TSH receptors on the surface of thyrocytes contain N-glycans, which are crucial to their proper activity. Thyroglobulin (Tg), the protein backbone for synthesis of thyroid hormones, is a heavily N-glycosylated protein, containing 20 putative N-glycosylated sites. N-oligosaccharides play a role in Tg transport into the follicular lumen, where thyroid hormones are produced, and into thyrocytes, where hyposialylated Tg is degraded. N-glycans of the cell membrane transporters sodium/iodide symporter and pendrin are necessary for iodide transport. Some changes in glycosylation result in abnormal activity of the thyroid and alteration of the metabolic clearance rate of hormones. Alteration of glycan structures is a pathological process related to the progression of chronic diseases such as thyroid cancers and autoimmunity. Thyroid carcinogenesis is accompanied by changes in sialylation and fucosylation, β1,6-branching of glycans, the content and structure of poly-LacNAc chains, as well as O-GlcNAcylation, while in thyroid autoimmunity the main processes affected are sialylation and fucosylation. The glycobiology of the thyroid gland is an intensively studied field of research, providing new data helpful in understanding the role of the sugar component in thyroid protein biology and disorders.

Perigestational exposure to low doses of PBDE-47 induces excessive ER stress, defective autophagy and the resultant apoptosis contributing to maternal thyroid toxicity

Brominated flame retardant 2,2′,4,4′‑tetrabromodiphenyl ether (PBDE-47) is known to induce developmental neurotoxicity by disturbing thyroid hormones (THs). Evidence shows that maternal THs are crucial for brain development and growth of fetuses and infants. However, little is known about the effects of PBDE-47 on maternal thyroid status and its mode of action. Here, using female Sprague-Dawley rats orally exposed to low doses of PBDE-47 (0.1, 1.0, 10 mg/kg/day) from pre-pregnancy until weaning of offspring to mimic human exposure, we show that perigestational exposure to PBDE-47 elevated serum triiodothyronine and thyroxine levels in mother rats. This is accompanied by disrupted thyroid follicle structure including expanded follicles, hyperplastic epithelial cells and shed cell remnants filled in the exhausted follicular lumen. Mechanistically, PBDE-47 enhanced apoptosis in thyroid tissue, as demonstrated by Caspase-3 activation, PARP cleavage and DNA fragmentation. Further study identified that PBDE-47 upregulated the levels of GRP78, ATF4, active Caspase-12 and CHOP, suggesting endoplasmic reticulum (ER) stress and unfolded protein response activation. Moreover, PBDE-47 reduced the levels of LC3-II, an autophagy marker protein essential for the autophagosomes formation, while increased the autophagy substrate p62 accumulation, indicating autophagy defect. Importantly, the colocalization of apoptotic cells with CHOP, a key mediator of ER stress-induced apoptosis, or p62, uncovered the contribution of excessive ER stress and defective autophagy to apoptosis. Collectively, our results suggest that excessive ER stress, defective autophagy and the resultant apoptosis are implicated in maternal thyroid injury following perigestational PBDE-47 exposure, which offers insight into a better understanding of PBDE-47-induced maternal thyroid toxicity.

Прогностичні маркери проліферації та апоптозу в пацієнтів на вузлові форми зоба на фоні автоіму-нного тиреоїдиту

Мета роботи: вивчити процеси апоптозу та проліферації в пункційному матеріалі щитоподібної залози (ЩЗ) при вузловому зобі на фоні автоімунного тиреоїдиту ВЗАІТ з використанням імуногістохімічного методу дослідження, а також визначити індекс проліферативної активності.Матеріали і методи. На пункційному матеріалі ЩЗ, отриманому від 75 хворих із гістологічно верифікованим діагнозом ВЗАІТ, проведено імуногістохімічне дослідження шляхом використання моноклональних антитіл проти Fas, FasL, Bcl-2, P53 та Ki67 антигенів.Результати досліджень та їх обговорення. Результати показали ступінь проліферативної активності тканини ЩЗ при ВЗАІТ. Виявлено високу проліферативну активність лімфоїдної тканини, помірну проліферативну активність тиреоцитів у ділянці лімфоїдної інфільтрації і низьку – поза нею. Виражена експресія Fas та FasL на тиреоцитах, у ділянках лімфоїдної інфільтрації побічно свідчить про те, що при ВЗАІТ відбуваються імунологічно зумовлені процеси апоптозу тиреоцитів. Збільшення кількості імунореактивних клітин, що експресують Ki67 у ділянці лімфоїдної інфільтрації і деструкції тиреоцитів, свідчить про регенерацію фолікулярного епітелію як компенсаторно-пристосувальну реакцію органа. Виражена експресія bsl-2 у лімфоцитах запобігає вхід клітин в апоптоз і подовжує час виживання клітин. Спостерігалася висока експресія протеїну р53 в ядрах тиреоцитів та фолікулярних просвітах, яку можна пояснити мутаціями гена р53, що дозволяє клітинам знайти толерантність до апоптичної дії ефекторів імунної системи.

Iodinated TG in Thyroid Follicular Lumen Regulates TTF-1 and PAX8 Expression via TSH/TSHR Signaling Pathway.

Our previous study showed that highly iodinated thyroglobulin (TG) inhibited thyroid transcription factor-1 (TTF-1) and paired box gene 8 (PAX8) expression, but the potential mechanism remains unclear. In this study, we constructed a thyroid follicle model in vitro to mimic its natural physiological structure and explored how iodinated TG in the follicular lumen tuned TTF-1 and PAX8 expression. Our data showed that lowly iodinated TG enhanced PKA activity while upregulation of both TTF-1 and PAX8 expression; and that highly iodinated TG triggered PKC activity while suppression of TTF-1 and PAX8 expression. Further, PKA agonist alone could increase TTF-1 and PAX8 expression while PKC agonist decreased TTF-1 and PAX8 level. If blocking PLC-PKC pathway using PKC-specific inhibitor, highly iodinated TG significantly promoted the expressions of TTF-1 and PAX8, and similarly PKA-specific blocker moderately inhibited TTF-1 and PAX8 expression. And opposite tendencies of TTF1 and PAX8 aberrant expression were observed in the condition of low iodinated TG when blocking PLC-PKC and cAMP-PKA signaling pathways. Our results indicated that iodinated TG manipulated TTF-1 and PAX8 expression through PLC-PKC and cAMP-PKA pathways, and highly iodinated TG played inhibitory role via PLC-PKC pathway from the TTF1 and PAX8 perspective while low level of iodinated TG was an activator through cAMP-PKA pathway. Our findings proved that iodinated TG in thyroid follicular lumen regulated TTF-1 and PAX8 expression through thyroid stimulating hormone/thyroid stimulating hormone receptor (TSH/TSHR) mediated cAMP-PKA and PLC-PKC signaling pathways. J. Cell. Biochem. 118: 3444-3451, 2017. © 2017 Wiley Periodicals, Inc.

Low Iodine in the Follicular Lumen Caused by Cytoplasm Mis-localization of Sodium Iodide Symporter may Induce Nodular Goiter.

Iodine is a key ingredient in the synthesis of thyroid hormones and also a major factor in the regulation of thyroid function. A local reduction of iodine content in follicular lumen leads to overexpression of local thyroid-stimulating hormone receptor (TSHr), which in turn excessively stimulates the regional thyroid tissue, and result in the formation of nodular goiter. In this study, we investigated the relationship between iodine content and sodium iodide symporter (NIS) expression by using the clinical specimens from patients with nodular goiter and explored the pathogenesis triggered by iodine deficiency in nodular goiter. In total, 28 patients were clinically histopathologically confirmed to have nodular goiter and the corresponding adjacent normal thyroid specimens were harvested simultaneously. Western blot and immunohistochemistry were performed to assay NIS expression and localization in thyrocytes of both nodular goiter and adjacent normal thyroid tissues. NIS expression mediated by iodine in follicular lumen was confirmed by follicular model in vitro. Meanwhile, radioscan with iodine-131were conducted on both nodular goiter and adjacent normal thyroid. Our data showed that NIS expression in nodular goiter was significantly higher than that in adjacent normal tissues, which was associated with low iodine in the follicular lumen. Abnormal localization of NIS and lower amount of radioactive iodine-131 were also found in nodular goiter. Our data implied that low iodine in the follicular lumen caused by cytoplasm mis-localization of NIS may induce nodular goiter.

Thyroid hormone biosynthesis and release

Thyroid hormones (TH) 3,5,3′,5’- tetraiodothyronine or thyroxine (T4) and 3,5,3’- triiodothyronine (T3) contain iodine atoms as part of their structure, and their synthesis occur in the unique structures called thyroid follicles. Iodide reaches thyroid cells through the bloodstream that supplies the basolateral plasma membrane of thyrocytes, where it is avidly taken up through the sodium/iodide symporter (NIS). Thyrocytes are also specialized in the secretion of the high molecular weight protein thyroglobulin (TG) in the follicular lumen. The iodination of the tyrosyl residues of TG preceeds TH biosynthesis, which depends on the interaction of iodide, TG, hydrogen peroxide (H2O2) and thyroid peroxidase (TPO) at the apical plasma membrane of thyrocytes. Thyroid hormone biosynthesis is under the tonic control of thyrotropin (TSH), while the iodide recycling ability is very important for normal thyroid function. We discuss herein the biochemical aspects of TH biosynthesis and release, highlighting the novel molecules involved in the process.

In situ cellular level Raman spectroscopy of the thyroid.

We report a novel Raman spectroscopy method for in situ cellular level analysis of the thyroid. Thyroids are harvested from control and lithium treated mice. Lithium is used to treat bipolar disorder, but affects thyroid function. Raman spectra are acquired with a confocal setup (514 nm laser, 20 µm spot) focused on a follicular lumen. Raman peaks are observed at 1440, 1656, and 1746 cm-1, corresponding to tyrosine, an important amino acid for protein synthesis. Peaks are also observed at 563, 1087, 1265 and 1301 cm-1. With lithium, the tyrosine peaks increase, indicating tyrosine buildup. Raman spectroscopy can study the impact of many exogenous treatments on thyroid biochemistry.

A Case of Mammary-Analogue Secretory Carcinoma of the Parotid Gland

Salivary gland carcinoma was originally classified as acinic cell carcinoma because of its histological similarity to secretory breast carcinoma. However, in 2010, Skalove et al. characterized this tumor as mammary-analogue secretory carcinoma, because like secretory breast carcinoma, it often expresses the ETV6-NTRK3 fusion oncogene. We encountered a case of mammary-analogue secretory carcinoma arising from the parotid gland. This specific diagnosis was made because (1) a follicular variant was present, (2) physaliphorous cells were observed, (3) no zymogen granules were observed, (4) the mucoid discharge in the follicular lumen was D-PAS-positive, and (5) the tumor showed immunostaining characteristics consistent with those of mammary-analogue secretory carcinoma (e.g., positive for S-100 and adipophilin). No additional treatment was administered to the patient after the resection, as the tumor is known to show a low malignancy potential and the surgical margins were negative. Tumor recurrence or metastasis has not been observed until date.

Defective ciliogenesis in thyroid hürthle cell tumors is associated with increased autophagy.

Primary cilia are found in the apical membrane of thyrocytes, where they may play a role in the maintenance of follicular homeostasis. In this study, we examined the distribution of primary cilia in the human thyroid cancer to address the involvement of abnormal ciliogenesis in different thyroid cancers. We examined 92 human thyroid tissues, including nodular hyperplasia, Hashimoto's thyroiditis, follicular tumor, Hürthle cell tumor, and papillary carcinoma to observe the distribution of primary cilia. The distribution and length of primary cilia facing the follicular lumen were uniform across variable-sized follicles in the normal thyroid gland. However, most Hürthle cells found in benign and malignant thyroid diseases were devoid of primary cilia. Conventional variant of papillary carcinoma (PTC) displayed longer primary cilia than those of healthy tissue, whereas both the frequency and length of primary cilia were decreased in oncocytic variant of PTC. In addition, ciliogenesis was markedly defective in primary Hürthle cell tumors, including Hürthle cell adenomas and carcinomas, which showed higher level of autophagosome biogenesis. Remarkably, inhibition of autophagosome formation by Atg5 silencing or treatment with pharmacological inhibitors of autophagosome formation restored ciliogenesis in the Hürthle cell carcinoma cell line XTC.UC1 which exhibits a high basal autophagic flux. Moreover, the inhibition of autophagy promoted the accumulation of two factors critical for ciliogenesis, IFT88 and ARL13B. These results suggest that abnormal ciliogenesis, a common feature of Hürthle cells in diseased thyroid glands, is associated with increased basal autophagy.

Morphological and Functional Changes in the Thyroid Follicles of the Aged Murine and Humans.

BackgroundAlthough both thyroid histology and serum concentrations of hormones are known to change with age, only a few reports exist on the relationship between the age-related structural and functional changes of the thyroid follicles in both mice and humans. Our objectives were to investigate age-related histological changes of the thyroid follicles and to determine whether these morphological changes were associated with the functional activity of the follicles.MethodsThe thyroid glands of mice at 18 weeks and at 6, 15, and 30 months of age were histologically examined, and the serum levels of thyroid hormones were measured in 11-week-old and 20-month-old mice. Samples of human thyroid tissue from 10 women over 70 years old and 10 women between 30 and 50 years of age were analyzed in conjunction with serum thyroid hormone level.ResultsThe histological and functional changes observed in the thyroid follicles of aged mice and women were as follows: variable sizing and enlargement of the follicles; increased irregularity of follicles; Sanderson’s polsters in the wall of large follicles; a large thyroglobulin (Tg) globule or numerous small fragmented Tg globules in follicular lumens; oncocytic change in follicular cells; and markedly dilated follicles empty of colloid. Serum T3 levels in 20-month-old mice and humans were unremarkable.ConclusionsThyroid follicles of aged mice and women show characteristic morphological changes, such as cystic atrophy, empty colloid, and Tg globules.

Glycolysis-associated enzymes existing in the follicular lumen of the thyroid may interfere with energy metabolism.

Synthesis and storage of the thyroid hormone precursor, thyroglobulin (TG), occurs within the follicular lumen of the thyroid and the TG is then absorbed into cells for further processing before release into the blood. However, the mechanism of energy metabolism in the follicular lumen of the thyroid remains unknown. In the present study, the three dimensional structure of thyroid follicles was constructed using a primary culture of swine cells and the follicular protein was identified via matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Three glycolysis-associated enzymes, enolase, pyruvate kinase and phosphoglyceraldehyde dehydrogenase were identified in addition to TG. These results support the hypothesis that anaerobic glycolysis of glucose exists in the follicle and supports energy consumption for hormone synthesis.

The reproductive cycle of the sea urchin Lytechinus variegatus (Echinodermata: Echinoidea) in southern Brazil

Lytechinus variegatus (Lamarck) is a common echinoid found on the Atlantic coast of America from North Carolina to southern Brazil and their populations inhabit sub littoral shallow-water environments. This research examined some aspects of reproduction in the southern limit of distribution of the species. A total of 86 adult individuals of L. variegatus (50.8 to 77.0 mm test diameter) were manually collected from September 2000 to September 2001 along the unconsolidated bottoms at the eurihaline sector of Paranagua Bay (48° 19’ W - 25° 35’ S) for histological characterization of gametogenesis, sex ratio and gonadal index (GI) evaluation. This population showed a synchronicity in maturation and spawning events (autumn and winter respectively), with a 1:1 sex ratio (χ2 = 2.57, df = 7, p < 0.05). The GI increase was associated with storage of nutritional resources periods (resting stages and proliferation) that occurred between late winter and spring. Biometric analyzes indicated a greater investment in size and/or quantity of gametes between late spring and summer, when it was possible to observe mature sperm occupying on average 63.9 % of the follicular lumen and oocytes reaching values of 70.0 to 75.0 μm. Mature oocyte mean size is the highest diameter of those developed by other regular echinoids in the same latitude. However, the average diameter of mature oocytes of L. variegatus in the southern limit of its distribution was lower than the values recorded for the northern hemisphere.

Is Sodium Monocarboxylate Transporter (SMCT1) A Protein Involved in the Apical Iodide Transport?

SMCT1 is a sodium-coupled cotransporter of monocarboxylates and short-chain fatty acids, which has been found in the apical membrane of diverse epithelia such as colon, kidney, brain and thyroid. In thyroid follicular cells, SMCT1 originally was associated with passive apical iodide (I-) transport. However, this evidence was subsequently rejected, as Xenopus oocytes expression studies of SMCT1 demonstrated that this protein does not cotransport iodide or other inorganic anions. Interestingly, the role of SMCT1 in I- transport is now controversial, because an I- transport mode occurs at low extracellular sodium concentrations, such as that found in thyroid follicular lumen. Considering this discrepancy, here we have performed a comparative study between SMCT1 and the sodium iodide symporter (NIS) in order to find conserved sodium/iodide binding motifs between both proteins. Specifically, homology models of hSMCT1 and hNIS were built based on the crystal structure of vSGLT, a sodium/galactose transporter from Vibrio parahaemolyticus. The hNIS model predicts an iodide-binding pocket between the transmembrane TM3 and TM7 segments, where mutated residues previously were identified in patients with hypothyroidism. A similar iodide-biding pocket was identified in the hSMCT1 model involving some conserved residues in homologs of these proteins. Putative sodium binding sites were also identified in the two models, which comprise characteristic sequence motifs of sodium-dependent transporters. Finally, molecular dynamics simulations of the iodide-bound models were performed, demonstrating that conserved aromatic residues may be key in iodide coordination. Our results provide further evidence of homology between SMCT1 and NIS transporters and predict residues involved in binding sodium and iodide. Future work will be essential for determining the precise transport mechanism in these proteins.