Endocrine factors in the hypothalamic regulation of food intake in females: a review of the physiological roles and interactions of ghrelin, leptin, thyroid hormones, oestrogen and insulin.

Controlling energy homeostasis involves modulating the desire to eat and regulating energy expenditure. The controlling machinery includes a complex interplay of hormones secreted at various peripheral endocrine endpoints, such as the gastrointestinal tract, the adipose tissue, thyroid gland and thyroid hormone-exporting organs, the ovary and the pancreas, and, last but not least, the brain itself. The peripheral hormones that are the focus of the present review (ghrelin, leptin, thyroid hormones, oestrogen and insulin) play integrated regulatory roles in and provide feedback information on the nutritional and energetic status of the body. As peripheral signals, these hormones modulate central pathways in the brain, including the hypothalamus, to influence food intake, energy expenditure and to maintain energy homeostasis. Since the growth of the literature on the role of various hormones in the regulation of energy homeostasis shows a remarkable and dynamic expansion, it is now becoming increasingly difficult to understand the individual and interactive roles of hormonal mechanisms in their true complexity. Therefore, our goal is to review, in the context of general physiology, the roles of the five best-known peripheral trophic hormones (ghrelin, leptin, thyroid hormones, oestrogen and insulin, respectively) and discuss their interactions in the hypothalamic regulation of food intake.

Oestrogens in the mammalian brain: from conception to adulthood--a review.

Environmental and plant oestrogens have been identified as compounds that when ingested, disrupt the physiological pathways of endogenous oestrogen actions and thus, act as agonists or antagonists of oestrogen. Although the risks of exposure to exogenous oestrogens (ExEs) are subject to scientific debate, the question of how ExE exposure affects the central nervous system remains to be answered. We attempt to summarise the mechanisms of oestrogenic effects in the central nervous tissue with the purpose to highlight the avenues potentially used by ExEs. The genomic and rapid, non-genomic cellular pathways activated by oestrogen are listed and discussed together with the best known interneuronal mechanisms of oestrogenic effects. Because the effects of oestrogen on the brain seem to be age dependent, we also found it necessary to put the age-dependent oestrogenic effects in parallel to their intra- and intercellular mechanisms of action. Finally, considering the practical risks of human ExE exposure, we briefly discuss the human significance of this matter. We believe this short review of the topic became necessary because recent data suggest new fields and pathways for endogenous oestrogen actions and have generated the concern that the hidden exposure of humans and domestic animal species to ExEs may also exert its beneficial and/or adverse effects through these avenues.

Partial cloning and localisation of leptin and its receptor in the one-humped camel (Camelus dromedarius).

Based on the studies and results presented here, leptin and its receptor were expressed by adipose tissue, mammary alveolar epithelial cells, liver hepatocytes, and the lining epithelium of the bile duct of the one-humped camel (Camelus dromedarius). Our observations support the biological importance of leptin in the mammary gland as well as the likely local effect of leptin on the peripheral tissues. We suggest that there may be an association between hepatic leptin and the lipogenic activity of the liver in the dromedary camel.

Thyroid hormone metabolism in the brain of domestic animals.

The action of thyroid hormones in the brain is strictly regulated, since these hormones play a crucial role in the development and in the physiological functioning of the central nervous system. It has been shown by many authors that brain tissue represents a special site of thyroid hormone handling. A relative independence of this tissue of the actual thyroid status was shown by our research group in birds and mammals. Hypothyroid animals can maintain a close to normal level of triiodothyronine in the brain tissue for extended periods. This phenomenon is due to at least three regulating mechanisms. (1) Uptake of thyroid hormones is enhanced. It was shown that the uptake by the telencephalon of labelled triiodothyronine (T3) was much higher in thyroidectomized (TX) animals than in controls or in thyroidectomized and T3 supplemented ones. (2) Conversion of thyroxine into triiodothyronine is increased. One of the most important elements of this process is the adjustment of the expression and activity of the type II deiodinase of the brain to a higher level. Enzyme kinetic studies, expression of TRalpha and beta nuclear thyroid hormone receptors and--after cloning the chicken type II deiodinase--in situ hybridization studies clearly supported the central role of the conversion process. (3) The rate of loss of triiodothyronine from the brain tissue is slowed down under hypothyroid conditions as evidenced by our hormone kinetic studies.

Expression of leptin and its receptors in various tissues of ruminants.

The energy metabolism of domestic animals is under the control of hormonal factors, which include thyroid hormones and leptin. Leptin signals from the periphery to the centre. It is mostly produced in the white adipose tissue and informs the central nervous system (CNS) about the total fat depot of the body. Low and high levels of leptin induce anabolic and catabolic processes, respectively. Besides controlling the food uptake and energy expenditure leptin is also involved in regulation of the reproduction and the immune system. Leptin is produced in several tissues other than fat. In the present paper the leptin expression of ruminant (Egyptian water buffalo, cow, and one-humped camel) tissues are examined. The mammary gland produces leptin in each species investigated. The local hormone production contributes to milk leptin and most probably helps to maintain lactation. Considerable leptin receptor expression was observed in the milk-producing epithelial cells, which is the same cell type that produces most of the udder leptin. Based on the results tissues participating in production have an autoregulative mechanism through which tissues can be relatively independent of the plasma leptin levels in order to maintain the desired function.

Expression and localisation of leptin and leptin receptor in the mammary gland of the dry and lactating non-pregnant cow.

Leptin and leptin receptor were studied in the mammary gland of non-pregnant dry and lactating cows. Using RT-PCR it was demonstrated that leptin and its short (Ob-Ra) and long (Ob-Rb) receptor isoforms are expressed both in the dry and the lactating mammary gland tissue. Tissue distribution of leptin and its receptor mRNA transcripts were examined by in situ hybridisation, while the leptin protein was localised by immunohistochemistry. Although in situ hybridisation is semiquantitative, our morphological data suggest that the epithelial leptin mRNA expression of the lactating gland is higher than that of the dry gland. To compare the leptin mRNA levels between dry and lactating udders competitive PCR was used, which showed no difference in leptin expression for the whole mammary tissues. The lack of difference in total leptin mRNA levels is explained by the high adipose tissue content of the dry mammary gland. Leptin and its receptor transcripts are expressed mainly in the epithelial cells of lactating cows, while in dry mammary tissue the signal is found in the stromal tissues as well. The results provide additional evidence that locally produced leptin takes part in the regulation and maintenance of mammary epithelial cell activity.

Partial cloning and localization of leptin and leptin receptor in the mammary gland of the Egyptian water buffalo.

Originally an overall metabolic control was attributed to the leptin hormone, which is produced mainly by the adipose tissue. Recently, leptin gene expression was demonstrated in several additional peripheral tissues. Furthermore, several isoforms of leptin receptor were found both in the central nervous system and in the peripheral tissues. Using reverse transcription and polymerase chain reaction analysis we demonstrate that leptin is expressed both in the adipose tissue and in the lactating mammary gland tissue of Egyptian water buffalo. Our results show that, short and long isoforms of leptin receptor are expressed in buffalo mammary gland tissue. We have partially cloned the buffalo leptin and its short and long isoforms of receptor, which show a high sequence homology to previously published sequences of other mammalian species especially to that of other ruminants. Localization of leptin and its receptor mRNA transcripts, as determined by in situ hybridization procedure, revealed that leptin and its receptor transcripts are expressed specifically in the alveolar epithelial cells of the mammary gland. These morphological data support that leptin could also act as an autocrine and paracrine mediator for mammary gland metabolism and as a facilitator of alveolar epithelial cell activity during lactation.

Transcriptional regulation of iodothyronine deiodinases during embryonic development.

A single dose of chicken growth hormone (cGH) or dexamethasone acutely increases circulating T(3) levels in 18-day-old chicken embryos through a reduction of hepatic type III iodothyronine deiodinase (D3). The data in the present study suggest that this decrease in D3 is induced by a direct downregulation of hepatic D3 gene transcription. The lack of effect of cGH or dexamethasone on brain and kidney D3 activity, furthermore suggests that both hormones affect peripheral thyroid hormone metabolism in a tissue specific manner. Dexamethasone administration also results in an increase in brain type II iodothyronine deiodinase (D2) activity and mRNA levels that is also regulated at a transcriptional level. In contrast, however, cGH has no effect on brain D2 activity, thereby suggesting that either GH cannot pass through the blood-brain barrier in chicken or that cGH and dexamethasone regulate thyroid hormone deiodination by different mechanisms. In addition, the very short half-life of D2 and D3 (t(1/2)<1 h) in comparison with the longer half life of type I iodothyronine deiodinase (D1, t(1/2)>8 h), allows for D2 and D3 to play a more prominent role in the acute regulation of peripheral thyroid hormone metabolism than D1.

[Therapeutic solution for a combined endo-periodontal lesion. Case report]. / Kombinált endoparodontális elváltozás megoldása (Esetismertetés).

The endo-periodontal lesion may lead to diagnostic and therapeutic difficulties in general dental practice. In the present case endo-periodontal inflammation of the lower left first molar caused the patient's complaints. The inflammation of periodontal and pulpal origin was separated although they simultaneously were present in the same time. Endo-periodontal lesion can be treated by endodontic and periodontal care and sometimes complemented by surgery. Scaling and polishing, as well as root planing were performed following the endodontic treatment of distal root, then the tooth was dissected and the mesial root was removed. Finally the remained distal part of molar was used as a bridge abutment. Combined endo-periodontal lesion can be cured with appropriate treatment as root filling, periodontal treatment, supplemented with tooth dissection.

Characterization of the 5'-flanking and 5'-untranslated regions of the cyclic adenosine 3',5'-monophosphate-responsive human type 2 iodothyronine deiodinase gene.

The type 2 iodothyronine deiodinase (D2) catalyzes T4 activation. In humans, unlike rodents, it is widely expressed, and its action probably contributes to both intracellular and plasma T3 pools. We have isolated the 6.5-kb 5'-flanking region (FR) and the previously uncloned 553 nucleotides (nt) of the 5'-untranslated region (UTR) of hdio2. The 5'-UTR is complex, with three transcription start sites (TSS) (708, 31, and approximately 24 nt 5' to the ATG), an alternatively spliced approximately 300-nt intron in the 5'-UTR, and three short open reading frames 5' to the initiator ATG. The previously reported approximately 7.5-kb D2 messenger RNA (mRNA) is actually an approximately 7-kb doublet that is present in thyroid, pituitary, cardiac and skeletal muscle, and possibly brain, but with only the longer transcript in placenta. A canonical cAMP response element-binding protein-binding site is present at about 90 bp 5' to the most 5'-TSS. It accounts for the robust response of the 6.8-kb hdio2 5'-FR to protein kinase A. Forskolin increases D2 mRNA in human thyroid cells, which may explain the high D2 mRNA in Graves' thyroid and thyroid adenomas. The hdio2 gene structure and Northern blot results suggest that D2 expression is tightly controlled and tissue specific.

Cloning and expression of the chicken type 2 iodothyronine 5'-deiodinase.

The type 2 iodothyronine deiodinase (D2) is critical for the intracellular production of 3,5,3'-triiodothyronine from thyroxine. The D2 mRNA of higher vertebrates is over 6 kilobases (kb), and no complete cDNA clones have been reported. Using 5'- and 3'-rapid amplification of cDNA ends and two cDNA libraries, we have cloned the 6094-base pair full-length chicken D2 cDNA. The deduced protein is approximately 31 kDa and contains two in-frame UGA codons presumably encoding selenocysteine. One of these is in the highly conserved active catalytic center; the other is near the carboxyl terminus. Unusual features of the cDNA include a selenocysteine insertion sequence element approximately 4.8 kb 3' to the UGA codon in the active center and three short open reading frames in the 5'-untranslated region. The Km of D2 is approximately 1.0 nM for thyroxine, and the reaction is insensitive to inhibition by 6-n-propylthiouracil. Chicken D2 is expressed as a single transcript of approximately 6 kb in different brain regions and in the thyroid and lung. Hypothyroidism increases D2 mRNA in the telencephalon. Unlike in mammals, D2 mRNA and activity are expressed in the liver of the chicken, suggesting a role for D2 in the generation of plasma 3,5,3'-triiodothyronine in this species.

Regional expression of the type 3 iodothyronine deiodinase messenger ribonucleic acid in the rat central nervous system and its regulation by thyroid hormone.

Type 3 iodothyronine deiodinase (D3) is a selenoenzyme that inactivates thyroid hormone. It is necessary for T3 homeostasis in the central nervous system. D3 activity has been identified in many regions of the brain and parallels thyroid status, but the level at which it is regulated and its specific cellular locations are not known. We evaluated the effect of thyroid status on the expression of the D3 gene within the central nervous system using in situ hybridization histochemistry. D3 messenger RNA (mRNA) was identified throughout, but with high focal expression in the hippocampal pyramidal neurons, granule cells of the dentate nucleus, and layers II-VI of the cerebral cortex. In every region, D3 mRNA abundance was correlated with thyroid status. Four different D3 transcripts were identified by Northern analyses, with evidence for region-specific processing, and D3 mRNA increased 4- to 50-fold from the euthyroid to the hyperthyroid state. D3 mRNA was not detectable in hypothyroid brain. In the central nervous system, the D3 gene is highly T3 responsive, and its focal localization within the hippocampus and cerebral cortex suggests an important role for T3 homeostasis in memory and cognitive functions.

The guanosine monophosphate reductase gene is conserved in rats and its expression increases rapidly in brown adipose tissue during cold exposure.

Non-shivering thermogenesis is required for survival of rodents during cold stress. Uncoupling protein-1 acts in brown adipose tissue (BAT) to transport protons, thus dissipating the proton gradient across the inner mitochondrial membrane. This permits respiration uncoupled from ATP synthesis. UCP-1 function is inhibited by the binding of purine nucleotides, with GTP/GDP being more potent than ATP/ADP. We used a cDNA subtraction analysis to identify cDNAs rapidly induced by cold exposure. One of these encodes rat guanosine monophosphate reductase (GMP-r). This was surprising in that previous data had suggested that this enzyme was absent in rodents. Rat GMP-r is 96% identical to human GMP-r, and its mRNA is increased 30-fold in BAT within 6 h of cold exposure. The gene is also expressed (but not cold-responsive) in muscle and kidney, but not in white fat. We speculate that the physiological function of the marked increase in BAT GMP-r during cold stress may be to deplete the brown adipocyte of guanine nucleotides, converting them to IMP, thus permitting enhanced UCP-1 function. This is a previously unrecognized regulatory aspect of thermogenesis, an essential physiological response of rodents to cold.

3,3',5-Triiodothyronine (T3) uptake and expression of thyroid hormone receptors during the adaptation to hypothyroidism of the brain of chicken.

Thyroid hormone action in the brain is strictly regulated, since these hormones play a crucial role in the development and physiological functioning of the central nervous system. Hormone kinetics and molecular events at the nuclear receptor level during the adaptation of the brain of chicken to hypothyroidism were simultaneously investigated. Data obtained by Oldendorff's 'single-pass' technique showed a significantly higher labelled 3,3'5-triiodothyronine (125I-T3) uptake into the brain of surgically thyroidectomized (TX) 2-week-old broilers after 1 week of surgery in comparison to sham-operated (SH) and t3 supplemented (TX + T3) controls in the 10th second after the bolus injection. Telencephalons showed the highest, while cerebellum the lowest uptake intensity in all groups. In a similar arrangement of experiments the expression of the TR alpha- and TR beta nuclear thyroid receptors in the telencephalon of TX and control chickens was investigated by a semiquantitative RT-PCR-based approach for beta-actin, then amplified for thyroid receptors. The level of both the TR alpha and TR beta coding mRNA was elevated in hypothyroidism. In conclusion, the presented hormone kinetics and TR expression data provide further details of the cellular and molecular events occurring during the adaptation to hypothyroidism of the brain of chicken.

Regional distribution of type 2 thyroxine deiodinase messenger ribonucleic acid in rat hypothalamus and pituitary and its regulation by thyroid hormone.

To identify the specific locations of type 2 deiodinase (D2) messenger RNA (mRNA) in the hypothalamus and pituitary gland and determine its regulation by thyroid hormone, we performed in situ hybridization histochemistry, Northern analysis, and quantitative RT-PCR in euthyroid, hypothyroid, and hyperthyroid rats. By in situ hybridization histochemistry, silver grains were concentrated over ependymal cells lining the floor and infralateral walls of the third ventricle extending from the rostral tip of the median eminence (ME) to the infundibular recess, surrounding blood vessels in the arcuate nucleus (ARC), and in the ME adjacent to the portal vessels and overlying the tuberoinfundibular sulci. Silver grains also accumulated over distinct cells in the midportion of the anterior pituitary. In hypothyroid animals, an increase in signal intensity was observed in the caudal hypothalamus, and a marked increase in the number of positive cells occurred in the anterior pituitary. Microdissection of the hypothalamus for Northern and PCR analysis established the authenticity of D2 mRNA in the caudal hypothalamus, and confirmed that the majority of D2 mRNA is concentrated in this region. The distribution of D2 mRNA suggests its expression in specialized ependymal cells, termed tanycytes, originating from the third ventricle. Thus, the tanycyte is the source of the high D2 activity previously found in the ARC-ME region of the hypothalamus. The results indicate that tanycytes may have a previously unrecognized integral role in feedback regulation of TSH secretion by T4.

Molecular biological and biochemical characterization of the human type 2 selenodeiodinase.

Type 2 deiodinase (D2) is a low K(m) iodothyronine deiodinase that catalyzes the removal of a single iodine from the phenolic ring of T4 or rT3. We sequenced and subcloned the open reading frame from a partial complementary DNA (cDNA) clone (2.1 kilobases) prepared by Genethon (Z44085) from a human infant brain cDNA library. The open reading frame encodes a putative 273-amino acid protein of 31 kDa with greater than 70% similarity to the Rana catesbeiana D2 protein. Transient expression of the cDNA produces a low K(m) (5 nM for T4; 8 nM for rT3) propylthiouracil- and gold thioglucose-resistant 5'-deiodinase in 293-HEK cells. Human D2, like human type 1 (D1) and type 3 (D3) deiodinases, is a selenoenzyme, as evidenced by 1) the presence of two in-frame UGA codons (positions 133 and 266), 2) the synthesis of a 31-kDa 75Selabeled protein in D2 cDNA-transfected cells, and 3) the requirement for a 3'-selenocysteine incorporation sequence element for its translation. Unlike D1 and D3, we were not able to covalently label overexpressed D2 with N-bromoacetyl [125I]T3 or -T4. We found that the human D2 messenger RNA is 7-8 kilobases and is expressed in brain, placenta, and, surprisingly, cardiac and skeletal muscle. Type 2 deiodinase activity was also present in human skeletal muscle. These results indicate that there are unique features of D2 that distinguish it from the two other selenodeiodinases. The expression of D2 in muscle suggests that it could play a role in peripheral, as well as intracellular, T3 production.

Effect of severe energy restriction and refeeding on thyroid hormones in bulls.

Fifty-three Holstein-Friesian breeding bulls (944.99 +/- 14.59 kg) were fasted for 4 weeks. The influence of feeding on thyroid hormones was studied by comparing a starting point with a 4-week fasting period and a refeeding period. Blood samples were taken via a jugular vein catheter at 8:00 a.m. one day before, then once every week during, and two times after the fasting period. Plasma thyroxine (T4) and triiodothyronine (T3) levels were determined by direct radioimmuno-assay. The concentration of T4 and T3 decreased during fasting. The concentration of T3 increased after refeeding, but that of T4 did not. These data suggest that fasting is associated with a decrease in the peripheral conversion of T4 to T3 and, consequently, less T4 is converted into T3.

Kinetic parameters of plasma thyroid hormone and thyroid hormone receptors in a dwarf and control line of chicken.

Sex-linked dwarf chickens have in plasma low triiodothyronine (T3) levels and slightly raised thyroxine (T4) concentrations and are functional hypothyroid. The kinetic parameters of T4 and T3 were investigated using 125I-labeled hormones. In addition the nuclear T3-receptors in the liver were examined using a radioreceptorassay and Scatchard analysis. Four-week-old dwarf (dw) and normal (Dw) chickens were injected with 125I-labeled T3 or T4 and blood samples taken 60, 120, 180, and 300 min after 125I-T3 injection and 120, 240, 360, and 480 minutes after 125I-T4 treatment. Labeled T4 and T3 and the degradation products were separated by paper chromatography. After the paper strips were dried, the iodinated compounds were visualized and counted in the gamma counter. The kinetic parameters, the half-life time (T1/2), the apparent distribution volume (Vd) and the metabolic clearance rate (MCR) were calculated using the natural base logarithm values of the measured radioactivity plotted against time and used for linear regression. T4 was cleared from circulation more slowly in dwarf than in control chicks and reflected a longer T1/2 (21.8%) and a reduced MCR (45%). The Vd tended to be lower (34.7%) in dwarfs. While the T1/2 of T3 was longer (28.1%) in dwarf chickens than in control animals, the MCR for T3 was considerably increased (31.8%). This results from an increased Vd (63.1%) in the dwarf chicks. The T3-receptor study in the liver of dwarf and non-dwarf chickens from Week 1 to Week 4 posthatching revealed that the total capacity and the affinity constant of the binding sites were comparable in dwarf and normal chickens. However, the occupancy of the receptors was higher in the dwarf animals.

Increased sensitivity to triiodothyronine (T3) of broiler lines with a high susceptibility for ascites.

1. In the studies reported here, broiler lines divergently selected for susceptibility to ascites under low temperature conditions were tested for their sensitivity to 3,3',5-triiodothyronine (T3) with respect to growth rate, rate of mortality, plasma concentrations of T3, right ventricular hypertrophy (RVH) and incidence of ascites. 2. Mean body weight of the ascites-susceptible line (BC-line) was higher than that of the ascites-resistant line (A-line). Adding 0.5 mg T3/kg of the diet depressed growth rate to the same extent in both lines. The effect of T3 on growth was more pronounced for males than for females. 3. T3-supplementation increased the relative weight of the heart and the incidence of RVH to the same extent in both lines. More of the T3-treated BC-line chickens had fluid accumulation in the abdominal cavity than the T3-treated A-line chickens. 4. Dietary T3-treatment depressed the plasma concentration of growth hormone (GH) profoundly and insulin-like growth factor (IGF-I) slightly but to the same extent in both lines. The coefficient of variation of GH concentrations indicate that T3 treatment mainly decreased GH-pulsatility in young growing broilers. 5. Higher doses of dietary T3 (1 and 2 mg/kg) increased mortality in a dose-dependent manner. With 2 mg T3/kg, mortality in the BC-line was almost double that in the A-line. 6. These studies indicate that the development of ascites could be linked with thyroid function. Moreover, dietary T3 supplementation could be used to help identify ascites-inducing factors or genetic lines with differential sensitivity for ascites.

Elimination and metabolism of triiodothyronine depend on the thyroid status in the brain of young chickens.

The hypothyroid chicken brain has been found to preserve more triiodothyronine (T3) than expected from plasma T3 levels. A possible explanation is that the elimination of T3 from the hypothyroid brain is decreased. In the present experiments, the elimination rate of T3 was compared in surgically thyroidectomized animals and sham-operated controls. It was found that both T3 coming from the plasma and T3 derived locally in the cells from thyroxine have a significantly lower elimination rate in thyroidectomized chickens than in sham-operated ones. Therefore it is concluded that the adaptation of the brain to hypothyroid conditions is partly regulated by reducing the loss of the active thyroid hormone (i.e. T3) via metabolic and tissue-to-plasma exchange.