Molecular cloning of the cDNA encoding the β subunit of thyrotropin and regulation of its gene expression by thyroid hormones in the goldfish, Carassius auratus

A cDNA encoding the subunit of thyrotropin (TSH) was isolated from a goldfish (Carassius auratus) pituitary gland cDNA library. By comparing the sequence with other teleost TSHs, a signal peptide of 19 amino acids and a mature hormone of 131 amino acids were predicted for goldfish TSH subunits. The resulting putative mature hormone of 131 amino acids had well-conserved cysteine positions and a putative N-linked glycosylation site; homology was 51–67% with TSHs from other teleosts, 38–43% with tetrapod TSHs, but only 27 and 29% with goldfish GTH-I and -II, respectively. We also examined the effects of thyroid hormones (TH) and thiourea (TU, an inhibitor of TH production) treatments on TSH and GTH subunit gene expressions in the goldfish pituitary gland. After thyroxine (T₄) treatment, circulating T₄ concentration increased and TSH mRNA level decreased. Supressing the amount of circulating T₄ and triiodothyronine (T₃) by TU treatment increased the TSH mRNA level. Moreover, T₄ replacement therapy (simultaneous treatment of both TU and T₄) caused a high level of circulating T₄ and a low level of circulating T₃, and a decrease in the TSH mRNA level. Thus, changing levels of circulating TH exert a negative feedback on the level of TSH subunit mRNA in goldfish in vivo. On the other hand, GTH subunit mRNA levels were not affected by changes in the levels of circulating TH.     

幼龄山羊腺垂体分泌TSH活动的季节和昼夜变化

用RIA法测定幼龄去势莎能公山羊血清中TSH含量的季节和昼夜变化,发现血清中TSH的年平均含量为3.198±1.02μIU/ml。冬季昼夜平均含量最高,为4.454±0.34μIU/ml,春季、秋季、夏季昼夜平均含量分别为3.510±0.39μIU/ml,2.748±0.41μIU/ml,2.078±0.59μIU/ml。各季度血清中TSH含量,有着极明显的季节差异。全年平均含量的昼夜变化及各季度含量的昼夜变化,均无明显的昼夜差异,但血清中TSH含量有明显的个体差异。     

鸡胚胎腺垂体促甲状腺激素细胞的发生及其变化

应用免疫组织化学方法,对第3.5～20.5 d鸡胚腺垂体促甲状腺激素亚单位(TSH撸┫赴姆⑸⑹亢托翁植急浠媛山辛搜芯俊=峁允荆杭ε叻⒂牡?0.5 d,有少量TSH呦赴植加谙俅固迩耙叮赴派常伲舜螅患ε叻⒂牡?4.5 d以后,TSH呦赴灾黾樱≒ < 0.05),细胞体积增大,胞浆增多,细胞着色浓,分布于腺垂体前叶边缘区,少量分布于后叶背侧。研究结果证明,鸡胚腺垂体TSH呦赴⑸谂咛シ⒂闹衅冢幌赴脑鲋撤只谭⑸诩ε叻⒂闹泻笃冢籘SH的合成和分泌功能在胚胎发育的后期最旺盛。TSH呦赴植加谙俅固迩耙逗秃笠侗巢唷?     

Effect of recombinant human TSH on the uptake of radioactive iodine (I) by the thyroid gland in healthy beagles

In human medicine, recombinant human thyroid-stimulating hormone (rhTSH) increases thyroid radioactive iodine uptake (RAIU), allowing radioiodine-131 (¹³¹I) dose reduction and greater efficacy in the treatment of differentiated thyroid cancer and multinodular goiter. The goal of this study was to evaluate the effect of rhTSH, administered 24 h and 48 h before radioiodine-123 (¹²³I), on the thyroid RAIU in healthy dogs. Seven healthy euthyroid beagles were randomly allocated to 3 groups (2 groups of 2 dogs and 1 group of 3 dogs) in a prospective, blinded, crossover study. At Week 1, 1 group received ¹²³I for a baseline RAIU; 1 group received 100 μg of rhTSH IV 24 h before ¹²³I, and 1 group received 100 μg of rhTSH IV 48 h before ¹²³I. All dogs received 37 MBq of radioactive ¹²³I IV, and thyroid RAIU was determined 8 h, 24 h, and 48 h thereafter. The study was designed in such a manner that each dog received the 3 treatments and a wash-out period of 3 wk was respected in between. Blood samples were taken for measurement of serum total thyroxine (TT4) and thyrotropin (TSH) concentrations at baseline and 6 h, 12 h, 24 h, and 48 h after rhTSH administration. Recombinant human TSH caused no significant change on thyroid RAIU. The overall mean thyroid RAIU significantly decreased during the study independent of the treatment. Recombinant human TSH significantly increased serum TT4 concentration, which peaked 6 h after rhTSH administration. Compared to baseline, serum TSH concentration remained higher at 6 h, 12 h, 24 h, and 48 h. However, a statistically significant difference was reached only at 6 h and 12 h after rhTSH administration. No adverse effects of rhTSH were observed during the study. Further studies are needed to determine the best timing and dosage of administration of rhTSH in healthy and thyroid carcinoma dogs.     

梅山猪垂体中GH、PRL及TSHβ基因发育性表达规律的研究

生长激素(GH)、催乳素(PRL)及促甲状腺激素β亚基(TSHβ)是动物生长发育过程中的重要调控因子。本研究我们通过RT-PCR检测了GH、PRL及TSHβ基因在不同生长阶段梅山猪垂体中mRNA的表达水平。结果发现:GH和TSHβ基因在公、母猪的生长发育过程中的表达量总体上都呈现先升高后降低的趋势;而PRL在公猪中的表达量在不同发育阶段相对稳定,而在母猪中呈现先下降后升高的趋势。总体上GH、PRL及TSHβ3个基因在公、母猪的生长发育过程中的表达呈现性别二相性。其中GH基因的表达在1、30、90和150日龄公、母猪间呈现出显著差异(P0.05);而PRL基因的表达在1、60和90日龄的公、母猪表现显著差异(P0.05);而TSHβ基因的表达在60和150日龄的公、母猪表现显著差异(P0.05)。     

Concentrations of thyroid hormones and iodothyronine binding proteins in serum of the koala (Phascolarctos cinereus)

Objective To examine circulating total and free thyroid hormone (T₃ and T₄) concentrations, determine serum iodothyronine binding characteristics and estimate thyroid stimulating hormone (TSH) activity in sera of coastal and inland koalas.
Design A prospective study.
Procedure Koala serum T₃ and T₄ were measured by radioimmunoassay. T₄ binding parameters were determined by radioligand binding and electrophoresis. Koala TSH values were determined by bioassay.
Results Mean total T₄ concentrations were 3.2 ± 2.1 nM although values were significantly higher in inland-dwelling females in comparison to coastal-dwelling males. Free T₄ was 3.3 ± 2.1 pM. Total and free T₃ were 0.4 ± 0.2 nM and 1.4 ± 0.9 pM respectively, although these values were at the lower end of the assay detection limit and should be viewed with reservation. Electrophoresis of [¹²⁵I]-T₄-labelled serum revealed only two proteins of electrophoretic mobility similar to human transthyretin (TTR) and albumin. Scatchard analysis of T₄ binding to serum gave a curvilinear plot, which could be resolved into two binding sites with affinities identical to that of TTR and albumin but both of low concentration. The bioactivity of the TSH present in the sera was measured using a cell line (JP09) transfected with the human TSH receptor. The mean level of stimulation found in the sera corresponded to a bovine TSH activity of less than 10 mU/L.
Conclusion These results suggest that the serum concentrations of free and total thyroid hormones in koalas are low compared to other marsupials and very low compared to eutherian mammals. The mechanism of maintenance of euthyroidism in this species remains to be determined.     

Clinical Evaluation of a Novel Liquid Formulation of L-Thyroxine for Once Daily Treatment of Dogs with Hypothyroidism

Background: A liquid solution of levothyroxine (L-T4) is available for treatment of canine hypothyroidism.
Hypothesis: Once daily oral administration of a liquid L-T4 solution is effective and safe for controlling hypothyroidism in dogs.
Animals: Thirty-five dogs with naturally occurring hypothyroidism.
Methods: Dogs received L-T4 solution PO once daily at a starting dosage of 20 μg/kg body weight (BW). The dose was adjusted every 4 weeks, based on clinical signs and peak serum total T4 (tT4) concentrations. Target peak serum tT4 and thyroid stimulating hormone (TSH) concentrations, 4–6 hours posttreatment, were 35–95 nmol/L and < 0.68 ng/mL, respectively. Dogs were followed for up to 22 weeks after establishment of the maintenance dose.
Results: Clinical signs of hypothyroidism improved or resolved in 91% of dogs after 4 weeks of L-T4 treatment at 20 μg/kg once daily. The maintenance dose was established in 76, 94, and 100% of dogs after 4, 8, and 12 weeks of treatment, respectively. This was 20 μg L-T4/kg BW for 79% of the dogs, 30 μg/kg BW for 15%, and 10–15 μg/kg BW in the remaining 6%, once daily. Thereafter, median peak tT4 and TSH concentrations were 51 nmol/L and 0.18 ng/mL, respectively, and remained stable during the 22-week follow-up; clinical signs did not recur.
Conclusions and Clinical Importance: All of the hypothyroid dogs had rapid clinical and hormonal responses to supplementation with the PO-administered L-T4 solution. The starting dosage of 20 μg L-T4/kg BW once daily was suitable for 79% of dogs.     

Thyrotropin-releasing hormone-induced growth hormone secretion in dogs with primary hypothyroidism

Primary hypothyroidism in dogs is associated with increased release of growth hormone (GH). In search for an explanation we investigated the effect of intravenous administration of thyrotropin-releasing hormone (TRH, 10 microg/kg body weight) on GH release in 10 dogs with primary hypothyroidism and 6 healthy control dogs. The hypothyroid dogs had a medical history and physical changes compatible with hypothyroidism and were included in the study on the basis of the following criteria: plasma thyroxine concentration < 2 nmol/l and plasma thyrotropin (TSH) concentration > 1 microg/l. In addition, (99m)TcO(4)(-) uptake during thyroid scintigraphy was low or absent. TRH administration caused plasma TSH concentrations to rise significantly in the control dogs, but not in the hypothyroid dogs. In the dogs with primary hypothyroidism, the mean basal plasma GH concentration was relatively high (2.3+/-0.5 microg/l) and increased significantly (P=0.001) 10 and 20 min after injection of TRH (to 11.9+/-3.5 and 9.8+/-2.7 microg/l, respectively). In the control dogs, the mean basal plasma GH concentration was 1.3+/-0.1 microg/l and did not increase significantly after TRH administration. We conclude that, in contrast to healthy control dogs, primary hypothyroid dogs respond to TRH administration with a significant increase in the plasma GH concentration, possibly as a result of transdifferentiation of somatotropic pituitary cells to thyrosomatotropes.     

冷应激对雏鸡下丘脑-垂体-甲状腺轴的影响

越来越多的证据表明,应激能够激活下丘脑-垂体-甲状腺轴,进而影响促甲状腺激素释放激素(TRH)、促甲状腺激素(TSH)及甲状腺激素的合成与分泌。为了探明冷应激对雏鸡下丘脑-垂体-甲状腺轴的影响,以公雏鸡为实验动物,进行急性(0.25、1、3、6、12h与24h)与慢性(5、10d与20d)冷应激处理(12±1)℃,检测了雏鸡下丘脑TRH mRNA的表达水平,血清TSH、FT(3T3的游离形式)及FT(4T4的游离形式)的含量。结果表明:急性应激时,TRH mRNA的表达水平在各应激时间点均显著升高,TSH变化不明显,FT3开始无变化,在6h时突然降低,而后又显著升高,FT4开始变化不大,6h后显著升高;慢性应激时,TRH mRNA的表达水平与相应的对照组相比显著降低,TSH变化仍不明显,FT3呈上升趋势,而FT4呈下降趋势。这说明冷暴露可以使雏鸡下丘脑-垂体-甲状腺轴的激素分泌发生改变,而且不同程度的冷应激对同一激素也会产生不同的影响。