Evidence of the Presence of Thyroid Hormones in Achatina fulica Snails

ABSTRACT The objective of this study was to identify thyroid hormones and to examine their putative site of synthesis in Achatina fulica snails. For this purpose, radioimmunoassays were performed for T3 and T4 before and after long starvation with or without hemolymph deproteinization. Sodium/iodide symporter activity in vivo was analyzed through 125I administration with and without KClO4 pretreatment. Only T4 was detected, and its concentration decreased due to starvation or deproteinization. However, high-performance liquid chromatography analysis also showed the presence of T2 and T3 apart from T4, but rT3 was not detected in the A. fulica hemolymph. The sodium/iodide symporter activity was greater in cerebral ganglia than digestive gland, but KClO4 treatment did not inhibit iodide uptake in any of the tissues analyzed. Altogether, our data confirm for the first time the presence of thyroid hormones in A. fulica snails and suggest their participation in the metabolism control in this species, although the putative site of hormone biosynthesis remains to be elucidated.

Ca2+ is a Regulator of the WNK/OSR1/NKCC Pathway in a Human Salivary Gland Cell Line

Wnk kinase maintains cell volume, regulating various transporters such as sodium-chloride cotransporter, potassium-chloride cotransporter, and sodium-potassium-chloride cotransporter 1 (NKCC1) through the phosphorylation of oxidative stress responsive kinase 1 (OSR1) and STE20/SPS1-related proline/alanine-rich kinase (SPAK). However, the activating mechanism of Wnk kinase in specific tissues and specific conditions is broadly unclear. In the present study, we used a human salivary gland (HSG) cell line as a model and showed that Ca2+ may have a role in regulating Wnk kinase in the HSG cell line. Through this study, we found that the HSG cell line expressed molecules participating in the WNK-OSR1-NKCC pathway, such as Wnk1, Wnk4, OSR1, SPAK, and NKCC1. The HSG cell line showed an intracellular Ca2+ concentration ([Ca2+]i) increase in response to hypotonic stimulation, and the response was synchronized with the phosphorylation of OSR1. Interestingly, when we inhibited the hypotonically induced [Ca2+]i increase with nonspecific Ca2+ channel blockers such as 2-aminoethoxydiphenyl borate, gadolinium, and lanthanum, the phosphorylated OSR1 level was also diminished. Moreover, a cyclopiazonic acid-induced passive [Ca2+]i elevation was evoked by the phosphorylation of OSR1, and the amount of phosphorylated OSR1 decreased when the cells were treated with BAPTA, a Ca2+ chelator. Finally, through that process, NKCC1 activity also decreased to maintain the cell volume in the HSG cell line. These results indicate that Ca2+ may regulate the WNK-OSR1 pathway and NKCC1 activity in the HSG cell line. This is the first demonstration that indicates upstream Ca2+ regulation of the WNK-OSR1 pathway in intact cells.

Gitelman syndrome combined with complete growth hormone deficiency

Gitelman syndrome is a rare autosomal recessive hereditary salt-losing tubulopathy, that manifests as hypokalemic metabolic alkalosis, hypomagnesemia, and hypocalciuria. It is caused by mutations in the solute carrier family 12(sodium/chloride transporters), member 3 (SLC12A3) gene encoding the thiazide-sensitive sodium chloride cotransporter channel (NCCT) in the distal convoluted tubule of the kidney. It is associated with muscle weakness, cramps, tetany, vomiting, diarrhea, abdominal pain, and growth retardation. The incidence of growth retardation, the exact cause of which is unknown, is lower than that of Bartter syndrome. Herein, we discuss the case of an overweight 12.9-year-old girl of short stature presenting with hypokalemic metabolic alkalosis. The patient, on the basis of detection of a heterozygous mutation in the SLC12A3 gene and poor growth hormone (GH) responses in two provocative tests, was diagnosed with Gitelman syndrome combined with complete GH deficiency. GH treatment accompanied by magnesium oxide and potassium replacement was associated with a good clinical response.

A Case of Gitelman Syndrome

Gitelman's syndrome is an autosomal recessive disorder characterized by hypokalemic metabolic alkalosis, hypomagnesemia, and hypocalciuria that has recently been reported to be linked to thiazide-sensitive Na-Cl cotransporter gene mutation. We have experienced one patient whose initial complaint was paresthesia of hand and feet, who had hypokalemic metabolic alkalosis, hypomagnesemia, and hypocalciuria. We report the case of Gitelman's syndrome with a brief review of related literature.

Topologia do simportador tireoideano sódio/iodeto / Topology of the thyroid sodium-iodine symporter

A síntese de hormônios tireoideanos depende fundamentalmente da captação de iodo do meio extracelular para o interior do tireócito. Esse processo é mediado por uma glicoproteína transmembrânica denominada simportador sódio/iodeto, que transporta iodeto para o interior do tireócito, juntamente com dois íons sódio em um processo de cotransporte. Esse processo é orquestrado pelo potencial eletroquímico gerado pela bomba Na+/K+ ATPïase dependente. O simportador sódio/iodeto também está envolvido no transporte ativo de iodeto em tecidos extratireoideos, tais como glândulas salivares, mucosa gástrica e a mama em lactação. A alta capacidade de acumular iodeto pelo tireócito constitui a base do diagnóstico cintilográfico e também da terapêutica com radioiodo em situações de hiperfunção tireoidea, como, por exemplo, na doença de Graves. Algumas mutações no simportador sódio/iodeto geram prejuízo no transporte de iodeto para o tireócito, resultando em hipotireoidismo congênito; além disso, o simportador sódio/iodeto pode tornar-se alvo de imunocomplexos, como, por exemplo, nas doenças tireoideanas autoimunes. Finalmente, o estudo molecular do simportador sódio/iodeto apresenta importância em muitas áreas, que compreendem desde proteínas transportadoras até o diagnóstico e tratamento de cânceres em tecidos tireoidianos e extratireoideos. Este artigo objetivou descrever o simportador sódio/iodeto presente na glândula tireoide, destacando sua sequência de resíduos de aminoácidos, topologia e todos os demais aspectos pertinentes a sua estrutura e função. Foi desenvolvido através de revisão sistemática da literatura nacional e internacional pelo indexador Medline/PubMed, utilizando os unitermos: iodeto, tireoide, transportador, topologia, sequência de resíduos de aminoácidos e estrutura

The synthesis of thyroid hormones depends essentially on the uptake of iodide by thyrocytes, which is mediated by an intrinsic membrane glycoprotein, the sodium-iodide symporter. The NIS actively cotransports a sodium cation and an iodide anion simultaneously. NIS-mediated transport of iodide is driven by the electrochemical sodium gradient generated by Na+/K+ ATPïase. Sodium-Iodide Symporter also mediates active iodide transport in other tissues, including salivary glands, gastric mucosa, and lactating mammary gland. The ability of the thyroid to accumulate iodide via NIS has long provided the basis for diagnostic scintigraphic imaging of the thyroid with radioiodine and served as an effective means for therapeutic doses of radioiodide to target and destroy hyperfunctioning thyroid tissue, as seen in Graves? disease. Another relevant clinical aspect of Sodium-Iodide Symporter is the fact that some spontaneous mutations have been identified as the cause of congenital iodide transport defect, resulting in hypothyroidism. Furthermore, the sodium-iodide symporter can become the target of autoantibodies, resulting in autoimmune thyroid diseases. Finally, the molecular analysis of NIS clearly holds the potential of having an even greater impact on a wide spectrum of s, ranging from the structure and function of transport proteins to the diagnosis and treatment of cancer, in thyroid and nonthyroid tissues. The aim of this paper is to describe the sodium/iodide symporter present in the thyroid gland, highlighting its sequence of amino acid residues, topology, and all other relevant aspects of structure and function. This study is based on a systematic review of the domestic and international literature found in Medline/ PubMed with the keywords: iodide, thyroid, carrier, topology, sequence of amino acid residues and structure

Water and Sodium Regulation in Heart Failure

Heart failure is the pathophysiological state characterized by ventricular dysfunction and associated clinical symptoms. Decreased cardiac output or peripheral vascular resistance lead to arterial underfilling. That is an important signal which triggers multiple neurohormonal systems to maintain adequate arterial pressure and peripheral perfusion of the vital organs. The kidney is the principal organ affected when cardiac output declines. Alterations of hemodynamics and neurohormonal systems in heart failure result in renal sodium and water retention. Activation of sympathetic nervous system, renin-angiotensin-aldosterone system and non-osmotic vasopressin release stimulate the renal tubular reabsorption of sodium and water. Dysregulation of aquaporin-2 and sodium transporters also play an important role in the pathogenesis of renal sodium and water retention.

Altered Regulation of Renal Sodium Transporters in Salt-Sensitive Hypertensive Rats Induced by Uninephrectomy

Uninephrectomy (uNx) in young rats causes salt-sensitive hypertension (SSH). Alterations of sodium handling in residual nephrons may play a role in the pathogenesis. Therefore, we evaluated the adaptive alterations of renal sodium transporters according to salt intake in uNx-SSH rats. uNx or sham operations were performed in male Sprague-Dawley rats, and normal-salt diet was fed for 4 weeks. Four experimental groups were used: sham-operated rats raised on a high-salt diet for 2 weeks (CHH) or on a low-salt diet for 1 week after 1 week's high-salt diet (CHL) and uNx rats fed on the same diet (NHH, NHL) as the sham-operated rats were fed. Expression of major renal sodium transporters were determined by semiquantitative immunoblotting. Systolic blood pressure was increased in NHH and NHL groups, compared with CHH and CHL, respectively. Protein abundances of Na+/K+/2Cl- cotransporter (NKCC2) and Na+/Cl- cotransporter (NCC) in the CHH group were lower than the CHL group. Expression of epithelial sodium channel (ENaC)-gamma increased in the CHH group. In contrast, expressions of NKCC2 and NCC in the NHH group didn't show any significant alterations, compared to the NHL group. Expressions of ENaC-alpha and ENaC-beta in the NHH group were higher than the CHH group. Adaptive alterations of NKCC2 and NCC to changes of salt intake were different in the uNx group, and changes in ENaC-alpha and ENaC-beta were also different. These altered regulations of sodium transporters may be involved in the pathogenesis of SSH in the uNx rat model.

Expression of Occludin in Porcine Renal Epithelial Cells / 대한해부학회지

Occludin is a cell adhesion molecule that is abundantly expressed in the kidney. However, the expression pattern in various renal epithelial cells is not well established. The purpose of this study was to determine the cellular localization along the tubular epithelial cells in the kidney. Kidneys from adult pigs crossbred of Yorkshire, Landrace and Duroc (three breeds) were processed for immunohistochemistry. Thiazide sensitive sodium chloride cotransporter (TSC), Na+-KATPase bat1, calbindinD28k, and H+-ATPase were used to identify the thick ascending limb, distal convoluted tubule, connecting tubule, and collecting duct, respectively. In the pig kidney, occludin was expressed in the apical domain of the tubular epithelial cells. The immunoreactivity of occludin was strongest in the collecting duct, and then gradually decreased in the connecting tubule, distal convoluted tubule, and thick ascending limb. Occludin expression was weak in the thin limbs of the loop of henle and in the proximal tubule in the pig kidney. These results suggest that occludin may be a major adhesion molecule in distal tubular epithelial cells and play a critical role in maintaining epithelial polarity of these nephron segments.

Association of thiazide-sensitive Na+-Cl* cotransporter gene polymorphisms with the risk of essential hypertension / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the association of thiazide-sensitive Na+ -Cl* cotransporter (TSC) gene 1784C/T and 2736G/A polymorphisms with the risk of essential hypertension (EH) in a Han nationality population.</p><p><b>METHODS</b>A community-based, case-control study including 190 EH patients and 94 sex- and age-matched controls was conducted. Genotypes of TSC gene 1784C/T and 2736G/A polymorphisms were analyzed by gene chip technology.</p><p><b>RESULTS</b>The genotype (1784C/T CC, CT, TT:87, 88, 15 vs 36, 52, 6û2736G/A GG, AG, AA:167, 22, 1 vs 83, 10, 1) and alleles frequency (1784C/T C, T:68.9%, 31.1% vs 66.0%, 34.0%; 2736G/A G,A:93.7%, 6.3% vs 93.6%, 6.4%) distribution of 1784C/T and 2736G/A showed no significant difference between the EH group and the control group (P >0.05). Moreover, no significant difference was observed in the frequencies distribution of four haplotypes (P > 0.05); Logistic regression analysis of haplotypes showed that the risk of EH had no significant difference in the population with different haplotypes (P > 0.05).</p><p><b>CONCLUSION</b>The 1784C/T and 2736G/A polymorphisms of TSC gene may not play an important role in the etiology of EH in a Han nationality population. The studies in the future are warranted to validate our findings.</p>

Increased Expression of Sodium Transporters in Rats Chronically Inhibited of Nitric Oxide Synthesis

The present study was done to determine whether endogenous nitric oxide (NO) plays a role in the regulation of sodium transporters in the kidney. Male Sprague-Dawley rats were treated with NG-nitro-L-arginine methyl ester (L-NAME, 100 mg/L drinking water) for 4 weeks. Control rats were supplied with tap water without drugs. Expression of Na, K-ATPase, type 3 Na/H exchanger (NHE3), Na/K/2Cl cotransporter (BSC1), and thiazide-sensitive Na/Cl cotransporter (TSC) proteins was determined in the kidney by Western blot analysis. Catalytic activity of Na,K-ATPase was also determined. The treatment with L-NAME significantly and steadily increased the systemic blood pressure. Total and fractional excretion of urinary sodium decreased significantly, while creatinine clearance remained unaltered. Neither plasma renin activity nor aldosterone concentration was significantly altered. The alpha1 subunit expression and the catalytic activity of Na, K-ATPase were increased in the kidney. The expression of NHE3, BSC1 and TSC was also increased significantly. These results suggest that endogenously-derived NO exerts a tonic inhibitory effect on the expression of sodium transporters, including Na, K-ATPase, NHE3, BSC1, and TSC, in the kidney.

A Case of Gitelman's Syndrome Presented with Chest Pain and Syncope / 대한신장학회잡지

Gitelman's syndrome is a rare autosomal recessive, inherited renal tubular disorder, first described by Gitelman et al. in 1966, and it is characterized by hypokalemic metabolic alkalosis, hypomagnesemia, salt wasting, normal to low blood pressure and rather low urinary calcium excretion rates with elevated plasma renin activity. This syndrome is caused by inactivating mutation in the SLC12A3 gene coding for the thiazide-sensitive sodium chloride cotransporter in the distal convoluted tubule. In most of the patients with Gitelman's syndrome, the disease manifests with transient episodes of muscular weakness and tetany in the adulthood. Herein, we report a case of Gitelman's syndrome atypically presented with chest pain and syncope.

Absence of intact thiazide-sensitive sodium-chloride cotransporter in the renal tissue of a Gitelman's syndrome patient / 대한내과학회지

BACKGROUND: Gitelman's syndrome is an autosomal recessive renal tubular disorder characterized by hypokalemic metabolic alkalosis, hypomagnesemia, and hypocalciuria. It is known to be caused by a mutation of SLC12A3 gene coding the sodium-chloride cotransporter (NCCT) in the distal tubule. The defect of NCCT in human renal tissues has not been investigated, and we tested whether the defect of NCCT can be detected in renal tissue of a patient with Gitelman's syndrome by using immunohistochemistry. METHODS: In an adult patient with Gitelman's syndrome, blood and urine samples were collected for measurement of biochemical parameters. Renal clearance study and gene analysis were performed. Immunohistochemistry was performed on the renal tissue of the patient using a rabbit polyclonal antibody directed against a synthetic peptide corresponding to a portion in the amino terminal tail for human NCCT. Normal human renal tissues from surgical nephrectomy due to renal cell carcinoma and renal biopsy tissues from patients with glomerulonephritis but without any electrolyte disturbance were used as controls. RESULTS: The patient had hypokalemic metabolic alkalosis, hypocalciuria and hypomagnesemia. Renal clearance study revealed a decrease in distal fractional chloride reabsorption after the administration of furosemide. SLC12A3 gene mutation (S967F) was found by direct sequencing method. Immunohistochemistry showed the absence of NCCT staining in the renal tissue of the patient. On the other hand, the immunostaining of other transporters was all positive in renal tissues from both Gitelman's syndrome patients and controls. CONCLUSIONS: We report the absence of intact NCCT in the renal tissue of a Gitelman's syndrome patient.

Attenuated Renal Excretion in Response to Thiazide Diuretics in Gitelman's Syndrome: A Case Report

Gitelman's syndrome is a variant of Bartter's syndrome characterized by hypocalciuria and hypomagnesemia. The administration of thiazide diuretics may induce a subnormal increase of urinary Na+ and Cl- excretion in patients with Gitelman's syndrome, consistent with the hypothesis that less Na+ and Cl- than normal is reabsorbed by the thiazide-inhibitable transporter in Gitelman's syndrome. Specific mutations of NaCl cotransporter, coupled with mutant NaCl cotransporter expression studies clearly demonstrated that many of the characteristics of individuals with Gitelman's syndrome are explained by lack of function of NaCl cotransporter. We recently diagnosed a patient with Gitelman's syndrome by performing the thiazide and furosemide tests, and it is suggested that the clearance studies by diuretic administration may be of diagnostic help in Gitelman's syndrome.