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Since 1928, the iodine concentration in the ovary has been known to be higher than in every other organs except the thyroid. The ovarian iodide uptake varies with sexual activities, is enhanced by estrogens and a hypothyroid state and blocked by goitrogens. The recent discovery of a sodium iodide symporter (NIS) in ovaries has offered a possible mechanism for ovarian iodide uptake and other functional similarities to its thyroid counterpart. Nevertheless, the physiological significance of ovarian iodine uptake and accumulation remains unknown. The presence of thyroid hormones (TH) in follicular fluid (FF) has been established recently. Our preliminary studies on TH in FF (1996-1998) in rabbits, pigs, horses showed that the concentration of T4 is generally lower than that in serum and that for T3 is within the normal range or higher. A positive correlation exists between the T4 levels in FF and serum but not between the corresponding T3 levels. These studies revealed, for the first time, the presence of the ovarian 5'-monodeiodinase system in FF capable of generating T3 (ovary-born T3) by outer ring deiodination of T4. In mares, seasonal polyestrus, ovarian 5'-monodeiodinase (MD) activity and FF T3 levels have been found to be higher during the ovulatory period than in the anovulatory one. The exact physiological significance of this system generating T3 and coexisting with isoforms of TH receptors in granulosa cells has not been elucidated. A direct role of T3 for the early follicular development, differentiation and for the steroidogenic capability of granulosa cells, although strongly suggested by data obtained from in vitro studies, has to be elucidated.

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The concentrations of thyroxine (T4), 3,3',5-triiodothyronine (T3) and 3,3',5'-triiodothyronine (reverse T3; rT3) in thyroid gland tissue and serum of the fetal pig (n = 68) from day 39 to 113 of gestation were measured. Tracer quantities of iodothyronines, displaying the onset of thyroid hormone activity, were found in the thyroid tissue on day 39, i.e. before the appearance of a measurable quantity of iodothyronines in the serum. The T4 and T3 thyroidal content showed the first rise between days 56 and 76. Then, T3 was increasing sharply from day 92 till birth, while T4 content was decreasing from about day 76 to a low value between day 92 and 105, and then showing an increase shortly before birth. The rT3 content was the highest on day 39 and then it was steadily decreasing to reach a nadir on about day 76. Measurable amounts of thyroid hormones (TH) in the serum were observed not earlier than on day 46 of gestation. Near birth, the tissues of the pig fetus are in a milieu characterised by the highest blood TH concentrations. The serum to thyroid concentration ratio for rT3 and T4 was generally below 1.0 until the last trimester of gestation, when it was over 5.0 for rT3 and over 4.0 for T4. By contrast, the T3 serum to thyroid ratio was below 0.5 throughout the gestation. The results show that the fetal pig thyroid displays a low rT3 and T4 content, but the marked T3 elevation observed near term supports the view that a high production and secretion of T3 near term may be a critical factor for normal postnatal adaptation to extrauterine cooling in the pig.

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The physiological significance of thyroid hormones (TH) present in colostrum and milk is still under consideration. The present study was aimed at determining milk thyroxine (T4) and triiodothyronine (T3) levels in three lactation phases (early, mid- and late) of the goat, and to measure activity of the milk 5'-deiodinase (5'-MD) enzyme responsible for the intramammary conversion of prohormone T4 to its metabolically highly active form T3. Thirty-two milk goats (Polish White breed) fed a standard diet were used for milk sampling. The highest TH levels in mammary secretion were recorded during the first 2-3 days post partum. Then the hormone levels decreased, and by about Day 7 fluctuated around the overall mean for the early-lactation phase (Days 1 to 24 of lactation), recording 0.134+/-0.059 microg T4 and 150.8+/-2.80 ng T3 in 100 ml of the milk. Such T4 concentrations appeared to be comparable to those in the rabbit and human, whereas the concentration of T3 was higher than in the cow, pig and mare's milk. Milk 5'-MD activity was higher (P < 0.01) during early and late lactation, compared to the mid-lactation phase. It coincided with low T4 and high T3 milk levels during early lactation, and with high milk T4 and low T3 concentrations during late lactation. The quantity of T4 and T3 available to newborn kids in milk suggests that TH ingested with the colostrum may have a physiological role during the early postnatal life of suckling goats.

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The effect of naturally acquired bacterial infection of the bovine udder on the activity of 5'-thyroxine monodeiodinase (5'-MD), and on the concentrations of the pro-inflammatory cytokines interleukin (IL)-6 and tumour necrosis factor (TNF)-alpha in milk, from healthy (control) and inflamed quarters, was determined. The diagnostic procedure included history and clinical examination of the udder, macroscopic evaluation of secretions, the Californian Mastitis Test, determination of somatic cell counts and bacteriological examination of milk. It has been found that the milk triiodothyronine (T3) content and the 5'-MD activity from inflamed quarters were decreased when compared with controls. The decrease in the milk T3 from subclinical mastitic quarters was manifested when somatic cell counts were > 10(6) ml(-1). TNF-alpha was on average 2-fold higher in infected milk, and the concentration of IL-6 was unchanged. These results suggest that the decreased T3 content in mammary secretions during naturally occurring mastitis is associated with the severity of inflammation, increased TNF-alpha concentration and impaired enzymatic activity of 5'-MD.

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Thyroid hormones are critical for structural and functional development of the testis and Sertoli cells are considered true target cells for triiodothyronine (T3). However, the role of thyroid hormones in the adult testis seems to be minimal and the mechanism by which they affect testicular function is not known. Due to the existing blood-testis barrier the concentration of thyroid hormones in seminal plasma is kept lower than in blood plasma. We have found that T3 may reach the testis not only from the circulation but also from local enzymic conversion of thyroxine to T3. The presence of the enzymic activity responsible for thyroxine 5'deiodination and for generating T3 locally was also found in boar's seminal plasma. The seminal plasma 5'-deiodinase (5'-D) appeared to be predominantly the propylthiouracil (PTU)-insensitive type II isoenzyme found, so far, in tissues where it plays a role in paracrine signalling. It contains selenocysteine in its molecule (inhibition by aurothioglucose), and has an apparent Km for reverse-T3 as substrate of 0.36 nM and a Vmax 23.8 fmol I-/mg protein/min. Because the seminal plasma 5'-D is partially, but uncompetitively, inhibited by PTU, the presence in seminal plasma of two 5'-D isoenzymes (type I and II) cannot be excluded. The 5'-D activity in testes increased significantly between week 3 and 4, and this increase was concomitant with increase in testicular size. The relationship between testicular weight gain and age showed a similar characteristic change and corresponded to the change in 5'-D activity. Unlike in rodents, the testis of the prepubertal pig has thyroid hormone receptors in Sertoli cells, and suggests that in growing piglets, testicular 5'-D is a key factor regulating local supply of biologically active T3, and is an essential factor in testicular paracrine function. The present results are the first demonstration and characterization of the 5'-deiodinase in seminal plasma.

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Thyroid hormones (TH) and somatotropin (ST) play critical role in lactation. One explanation of their multiple physiological actions is based on the functional interrelationships among ST, TH, and thyroxin deiodinase (5'D). This enzyme is present in the mammary tissue, milk cellular components, and whole milk and is responsible for intramammary production of triiodothyronine (T3). In rats in which the 5'D isozymes in the mammary gland and in the liver are similarly of type I (5'D-I), an enhancement of mammary 5'D-I causes a reduction of hepatic 5'D-I activities. This opposite rearrangement in the mammary and hepatic deiodinating activities is thought to be a factor of a homeorhetic response characterized by an increased and compartmentalized energy expenditure of the mammary gland. In the cow, the mammary 5'D is the type II (5'D-II) deiodinase. The 5'D-II, owing to its high catalytic efficiency, secures T3 production, making tissues relatively independent from the circulatory levels of TH and from variations in the hepatic 5'D-I activity. No significant alterations of 5'D-II isozymes were found during a low T3 syndrome. Location of tissue deiodinases in the cow, the 5'D-II in the mammary gland, and the 5'D-I in the liver make it so that T3 production in these two tissues can be dissociated in time to secure better local requirement for T3 supporting lactation. To date, attempts to evidence that the alterations in iodothyronines blood levels and in tissues' 5'Ds activity during lactation are due to ST action have not received clear experimental support in either cows or rats.

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It is generally accepted that hormones and tissue growth factors are supplied from mother to neonate via mammary secretion. Among the protein hormones, insulin and prolactin are considered as the most important milk components for neonates. The significance of the thyroid hormones, namely triiodothyronine (T3) generated locally by 5'-monodeiodinase (5'-MD) in the mammary tissues, for the mammary gland itself and for suckling neonates is still under consideration. In the present study the activity of the 5'-MD and the concentrations of T3 and insulin in mare's colostrum and milk during the first 21 days of lactation were measured. Post partum, T3 increased to its highest concentration around day 4 (1.14+/-0.08 nmol/L), then progressively decreased until day 7, reaching a relatively stable concentration of 0.71+/-0.06 nmol/L (overall mean for days 7-21). The colostral insulin concentration, highest on the day of parturition (401.0 +/-24.9 microU/mL), decreased to a nadir value (25.0+/-3.4 microU/mL) on day 5, after which it tended to increase. The mare's milk showed the presence of PTU-sensitive (type I) and PTU-insensitive (type II) 5'-monodeiodinases (5'-MD). Contrary to the classical type II 5'-MD, the mare's milk isoenzyme was inhibited non-competitively by aurothioglucose. A significant relationship (r=0.962, P < 0.01 ) between T3 concentration and 5'-MD activity, from the I st to the 6th lactational day was found, which may indicate a dependence of T3 concentration on the milk 5'-MD activity. The presence of 5'-MD of type II suggests that intra-mammary T3 generation may play a paracrine role supporting lactogenesis. Estimating that 1.8 microg of colostral T3 (0.456 microg/L) is consumed daily by a suckling foal, the T3 hormone action within the intestinal tract cannot be ruled out. This is the first paper to provide evidence of T3 and insulin concentrations, and of T4-5'-monodeiodinases activity in colostrum and milk of the mare.

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The response of type I 5'-monodeiodinase activity (5'-MD) to a s.c. injection or oral administration of melatonin was studied in 3-, 5-, and 7-day-old rabbits. Melatonin-treated animals showed higher activity of the type I 5'-MD in the liver and kidney and of type II 5'-MD in brown adipose tissue (BAT). This respond to melatonin treatment was age dependent. The stimulatory effect of melatonin on renal 5'-MD activity was observed only in 3- and 5-day-old rabbits and in the liver and BAT during the first week of life. Oral melatonin administration tended to exert a more marked effect on enzyme activity than s.c. injection of the hormone. Changes in 5'-MD activities were accompanied by an increase in serum iodothyronine (T4, T3, and rT3) concentrations. The T3 and rT3 increases may result from the deiodinating processes by the type I 5'-MD and 5-MD, respectively, whereas the rise in the serum T4 was probably due to the stimulatory effect of melatonin on the secretory activity of the thyroid gland itself. These results are the first description of the effects evoked by melatonin treatment during the early neonatal period in newborns of the altricial type.

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Previous work from this laboratory has shown that the thyroid gland of the fetal pig begins to function at about day 46-47 (0.40-0.415 fraction of gestational age). Sera from fetuses contain lower thyroxine (T4), 3,3',5-triiodothyronine (T3) and 3,3',5'-triiodothyronine (rT3) concentrations than maternal sera, except for about 2 weeks before term. The fetal T4 metabolism is dominated by the 5'-monodeiodinating activity (5'-MD). In the present study we measured the iodothyronines content, and the outer (5'-MD) and inner (5-MD) monodeiodinases activity, in homogenates of the placenta. The pig placenta, which is of the epitheliochorial type, was separated into the fetal and the maternal part. The concentrations of T4, T3 and rT3 were lower, and the deiodinating activity of 5'-MD and 5-MD higher, in the fetal than in the maternal placenta. The fetal placenta not only deiodinated more actively T4 to T3 and T4 to rT3, but degraded T3 to 3,3'-diiodothyronine (3,3'-T2) more actively than rT3 to 3,3'-T2. Such divergent deiodinating activity of T4 to T3, T3 to 3,3'-T2 and rT3 to 3,3'-T2 might favor establishing a relatively high and constant rT3 concentrations in fetal and maternal placentas, and a lower T3 in the fetal placenta. The inner ring deiodinating activity (excluding a day before parturition) was always more active in the fetal placenta, while the outer ring deiodinations varied in this respect, depending on the gestation stage. These results support the hypothesis that in the fetal pig, enzymatic deiodination of thyroid hormones forms a barrier which reduces transplacental passage of the hormones and that the fetal part of the placenta is the primary factor in the mechanism regulating the hormonal transfer. In spite of the presence of the barrier, there is an adequate maternal supply of thyroid hormones to the fetus in early gestation, which suggests that the enzymatic mechanism is influenced in some way by the thyroid status of the fetus.

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The aim of this study was to localize and measure the receptor-binding capacity for insulin in the stomach, duodenum, ileum, jejunum, caecum, colon and in isolated hepatocytes in 48 rabbits aged 1-7, 14 and 21 days. Two methods were employed: radioimmunoassay (RIA) and receptor-binding assay (RBA). Isolated rabbit hepatocytes were used to estimate the maximal binding (Bo). Insulin (INS) concentrations measured by RIA-INS test were highest in the colostrum (701 microU/mL as 100%) of the does, and in the blood (82 microU/mL) of newborn rabbits during the first day postpartum. The Bo of colostral insulin for isolated hepatocytes was 556 microU/mL, which corresponded to 79.1% of the total binding as assessed by the RIA-INS test. The Bo for insulin binding in the different parts of the digestive tract tested was the highest on the first day (mean 8.0 fM/g) and the lowest on the 6th day postnatally (mean 4.9 fM/g; P < 0.01). The receptor capacity for insulin varied in relation to the different parts of the digestive tract, and with the age of each individual rabbit.

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Thyroxine (T4), 3,3',5'-tri-iodothyronine (reverse T3; rT3) and di-iodothyronines (3,3'-T2 and 3',5'-T2) were measured in pig amniotic fluid (AF) and allantoic fluid (Al) between 32 and 113 days of normal pregnancy. Low but measurable quantities of T4 in AF and Al (2.1 +/- 0.3 and 3.2 +/- 0.5 nmol/l respectively) were found before the onset of fetal thyroid gland function, which indicates the maternal source of T4. The presence of rT3 (55.8 +/- 4.1 pmol/l in AF and 49.8 +/- 5.3 pmol/l in Al), 3,3'-T2 (45.5 +/- 0.6 pmol/l in AF and 49.2 +/- 9.2 pmol/l in Al) and 3',5'-T2 (20.8 +/- 2.6 pmol/l in AF and 24.0 +/- 2.2 pmol/l in Al) may be attributed to the monodeiodinase system already active in fetal pig tissues in early pregnancy, as demonstrated previously. T3 concentration was undetectable in both AF and Al. An approximately twofold increase in the levels of T4, rT3 and T2s in AF and Al at mid-gestation was observed. T4 and rT3 in AF showed a positive correlation with protein concentrations. AF rT3 concentration (but not T4) correlated with rT3 in the cord and maternal serum. The 3,3'-T2 and 3',5'-T2 in AF and Al showed parallel changes to rT3, while the rT3/3,3'-T2 and rT3/3',5'-T2 molar ratios remained constant. T4 concentrations in AF and Al were markedly lower than in corresponding maternal and fetal serum; the rT3 concentration in Al was equal to that in AF and two to four times lower than in fetal serum. In spite of differences between serum hormone patterns in the pig and human near term, iodothyronine concentrations in AF showed some similarities, mainly the following: undetectable T3, a strong correlation between rT3, T4 and AF total protein and the presence of 3,3'-T2 and 3',5'-T2 in measurable levels. Comparative data for Al, except the ones in the present study in the pig, are not available.

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On the basis of our studies which demonstrated that T3 is a natural milk-borne component of cow mammary secretions, this study investigated the influence of T3 (and thyroxine, T4) on the serum Ig level (used here as an indicator of intestinal absorption). Forty healthy calves were given a single dose of either T3 or T4 with the first colostrum meal 6 h following birth. Blood samples were taken before and 42-50 h after hormone administration. The T4 treatment resulted in metabolic changes that were reflected by an increase in blood glucose, triglycerides (TG), non-esterified fatty acids (NEFA), and a decrease in total serum proteins (TP). Ig levels were reduced from 27.5 milligrams (controls) to 20.6 milligrams. The T3 treatment caused an increase in serum TG and FFA (p < 0.01), and, in contrast to T4, an increase in TP and Ig (p < 0.001). These results indicated that peroral administration of T3, but not T4, could exert a positive effect on the transfer of immunolactoglobulins in the neonatal calf intestine. The contrasting hormonal effects are likely attributable to different responses of intestinal cells to T3 and T4.

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The protective effect of vitamin E and reduced glutathione (GSH) against lipid peroxidation in boar semen plasma was studied. The lipid peroxidation, measured by the test for thiobarbituric acid reactive substances (TBARS), doubled in the presence of the lipid peroxidation Fe(2+)-sodium ascorbate-inducing system. The ascorbate-induced TBARS were inhibited by about 62% through the water-soluble vitamin E analog (TROLOX) and about 57% by GSH. In the in vivo experiments, 7 wk of oral DL-alpha-tocopherol acetate (1000 IU/d/animal) administration caused a significant fall in the level of the semen plasma TBARS, from 2.2 +/- 0.09 to 1.2 +/- 0.13 nmol MDA/mL. The semen plasma superoxide dismutase (SOD) and GSSG tended to increase with the time of vitamin E administration, but the increment did not reach a significant level by the seventh week. The vitamin E supplementation significantly increased the number of spermatozoa per 1 cm3 of ejaculate. The protective role of vitamin E and GSH with respect to boar semen against fatty acid peroxidation and a positive influence of vitamin E supplementation on semen quality have been evidenced.

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The ontogeny of the fetal monodeiodinase systems, during the period before the onset of thyroid activity until birth, has been described only in the rat, rabbit and chick. We have studied 5'- and 5-monodeiodinase activities in the liver, kidney and placenta of pig fetuses, from day 32 of gestation up to birth, and on days 1, 3, 7 and 14 after birth. Fetuses (123) from 15 litters, 32 newborn piglets, 15 sows (mothers) and 10 non-pregnant pigs were used. The relationships between monodeiodinase activities, thyroxine (T4), tri-iodothyronine (T3) and reverse triiodothyronine (rT3) blood serum concentrations and sulphydryl groups (total (T-SH) and non-protein (NP-SH)), were measured. 5'-Monodeiodinase activity (5'-MD type I; converting T4 to T3) and 5-monodeiodinase (5-MD; converting T4 to rT3) were detectable in the liver at the onset of thyroid hormone biosynthesis, and before that time (day 32 of gestation) in the placenta. Liver 5'-MD activity was very low between days 32 and 84 of gestation, and then increased rapidly at about day 93, reaching a maximal value 2 days before term. Activity in the liver (but not in the kidney) was higher than that in maternal, newborn and non-pregnant adult tissue. The increase in NP-SH groups in the liver and kidney and T-SH groups in the placenta during the last 5 days of gestation coincided with the increase in 5'-MD activity.(ABSTRACT TRUNCATED AT 250 WORDS)

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A short review of the presence of hormones and tissues growth factors in milk during various stages of lactation. On comparative grounds, their possible physiological importance for the neonate is discussed.

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Simple diffusion or facilitated diffusion, by a carrier system represented by binding sites on plasma membranes, are thought to be responsible for thyroid hormones (TH) uptake by tissues. The concept implies the existence of an equilibrium ratio between plasma and cellular hormone content. Studies of TH presence in milk have shown that the passage of TH through mammary gland differs between T4, rT3, and T3, and only T3 exists in the milk in measurable amounts (without definite relationship between the levels of T3 in milk and the blood plasma). The present experiments have revealed that the whole milk or its cellular components (namely macrophages, lymphocytes and granulocytes) possess deiodinating enzyme system converting T4 into T3. The milk from animals displaying mastitis show lower T3 levels than controls. Of different enzyme systems, the outer ring deiodinase (5'D-II) dominates in mammary secretion. It plays an important role in galactopoiesis yielding biologically potent T3. The presence of TH deiodinating system(s) in mammary gland is thought to lower T4 and T3 in milk, already in small concentrations due to a poor permeability of the blood-mammary gland barrier for these compounds. On the other hand measurable amount of T3 in milk is due to a local enzyme mechanism generating T3 from T4. It has been concluded that the ultimate THs level in milk depends on the rate of transport of the hormones, their degradation and simultaneous generation of T3 from T4 in mammocytes.

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Both inner- and outer-ring iodothyronines deiodinating activity was found in homogenates of rabbit placentas. The T4 to rT3 and T3 to 3,3'-T2 deiodinating activity was already high on day 10 before delivery but decreased being about 7 times lowered on day 5. Once the T3 to 3,3'-T2 monodeiodination reached a low and a relatively steady level, the outer ring deiodination of T4 begun, reaching a peak value at about day 3 before term and then fell again. The fetal serum thyroid hormones levels were low, showing no significant variability during the period of observation. The results suggested that in the rabbit, representing animals in which the thyroid gland activity begins early in fetal life, there are two distinct phases of the placental monodeiodinating activity. The first is characterized by a high inner-ring deiodinating activity (yielding rT3) and is followed by the second phase with a high outer-ring deiodinating activity (yielding T3) declining just before term.

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Danowski et al. (1951) were probably the first who showed an increase of PBI in infants shortly after birth. Later a number of investigators presented further evidence on the increased thyroid gland function in newborns. This condition named "neonatal thyroid hyperactivity" [Fisher and Oddie, 1964] was described also in animals. The available data indicate that all the newborn mammals till now studied, independently of the maturation stage of development they reach at birth, display some features of thyroid hyperactivity, but in some it does not lead to hyperiodothyroninaemia. Interspecies differences coincide well with significance of the thyroid hormones for neonatal thermogenesis. There are few sequential studies of the three principal iodothyronines: T4, T3 and rT3 available at present. The most comprehensive data concern infants, newborn lambs and pigs. Immediately after birth, there is a sudden rise in serum thyroid hormone concentrations, with some species differences related to the degree of the increase and to the iodothyronines involved. The course of the postnatal hyperiodothyroninaemia is dependent on the maturation level reached at birth, food intake, and cooling relative to extrauterine environment. At least five main factors contribute to the immediate postnatal hyperiodothyroninaemia: 1) abrupt depletion of the preformed fetal hormonal iodine stores; 2) preferential T3 secretion; 3) increase in the T4 to T3 monodeiodination in the peripheral tissues; 4) a release of thyroid hormone content from peripheral reservoirs to plasma, and 5) action of other hormone(s) concomitantly released at birth. From the point of view of the thermal adaptability, the newborn mammals fall into two distinct groups: first, in which immediately after birth the metabolic rate decreases, and second, in which the metabolic rate increases, after cooling. Our understanding of the role and significance of hormonal factors involved in mechanisms of the postnatal thermogenesis is incomplete. However, some similarities in adaptation to cold in adults and in newborns seem to be relevant. Cold adaptation is accomplished by development of nonshivering thermogenesis (NST) depending on NA. The presence of brown adipose tissue (BAT) is essential for NA thermogenic action. According to the common opinion, based on data from laboratory animals and human baby, non-shivering thermogenesis, but not shivering, predominates in newborns. However, data from domestic animals indicate that shivering thermogenesis may be of comparable or greater thermogenic capacity than NST at birth. Besides, there are some newborns which have little or no BAT and associated NST.(ABSTRACT TRUNCATED AT 400 WORDS)

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Changes in the in vitro hepatic, kidney, skeletal muscle, heart and brain 5'- and 5-monodeiodinase activities were studied in piglets aged 4-12 h and 3, 7, 14, 21, and 42 days. As compared to the adult pig, the hepatic conversion of thyroxine (T4) to 3,5,3'-triiodothyronine (T3) and 3,3'5'-triiodothyronine (rT3) was on average 3-5 times higher in the liver, and equal or lower in the kidney. At birth, the liver generated about 66% of T3 and 75% of rT3, the other tissues the rest. Increasing contribution of skeletal muscles in the postnatal process of triiodothyronines generation was found. The apparent Km and Vmax values suggested higher capacity of T4 converting systems in newborns than in adults, and an increase in the enzyme affinities. Shortly after birth serum rT3 but not T3 corresponded to the level of hepatic and kidney T4 conversion.

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