Divergent deiodination of thyroid hormones in the separated parts of the fetal and maternal placenta in pigs.

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.

Ontogeny of the generation of diiodothyronines (3,3'-T2 and 3',5'-T2) from triiodothyronines in the liver and kidney during the fetal life of the pig.

The present study was conducted to obtain information on the generation of diiodothyronines from triiodothyronines (3,3'-T2 from T3, and rT3, and 3',5'-T2 from rT3) as a result of the activity of the tissue monodeiodinase enzymes (MD) in the liver and kidney of pig fetuses and their mothers between 32 and 113 days of gestation. T3-5-MD activity in the fetal kidney during the gestational period was stable and higher than in the liver and in the maternal kidney. In contrast, T3-5-MD activity of the liver was 3-4 times lower in fetal than in maternal tissue in the first half of pregnancy, and in the second half of pregnancy 3,3'-T2 production from T3 in the maternal liver was equal to, or lower than, that in the fetal liver. The activity of MD deiodinating rT3 (3,3'-T2 and 3',5'-T2 generation) increased significantly in fetal liver and kidney in the last 2-3 weeks of pregnancy and was higher than in maternal tissues. In both tissues examined the inner ring deiodinating activity (IRD) was 5-10 times lower as compared to the outer ring (ORD).

Amniotic and allantoic fluid concentrations of thyroxine, 3,3',5'-tri-iodothyronine, 3,3'-di-iodothyronine and 3',5'-di-iodothyronine in the pig during the period of gestation.

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.

Differences in the thyroxine, triiodothyronine and reverse triiodothyronine contents of fetal pig tissues relative to gestational age.

The concentrations of thyroxine (T4), 3,5,3'-triiodothyronine (T3) and 3,3',5'-triiodothyronine (reverse T3) were measured in the liver, kidney and skeletal muscles of pig fetuses from day 32 to 113 of age. The iodothyronines were found to be present already between days 32 and 39, that is before the onset of fetal thyroid function. In the liver, kidneys and skeletal muscles T4 reached maximal concentrations at midgestation (days 56 and 93). The T3 level increased with gestational age in all tissues examined but, as opposed to T4, its maximal value was reached shortly before birth. The rT3 level was the highest between days 85 and 100 in the liver and kidney; however, in the skeletal muscles it did not change with gestation. The presented profiles of change of T4 and rT3 in the examined tissues did not correspond to the blood hormone pattern already observed in pig, whereas changes in tissue T3 concentrations were similar to those in the serum.

Investigation of thyroxine monodeiodinase activity in the liver, kidney and brown adipose tissue of foetal and neonatal rabbit.

The maturation of the 5'- and 5-monodeiodinase system in liver, kidney and brown adipose tissue of rabbits, during the foetal period (from 21 days of gestation to birth) and the neonatal period (from birth to 3 weeks of life) was studied. A sudden increase of 5'- and 5-monodeiodinase activity in liver and kidney 3 days before birth was observed, falling to a nadir at day 3 after birth. Foetal and neonatal serum T4, T3 and rT3 concentration were very low and rose progressively with age, reaching adult values at about day 21. In the foetal brown adipose tissue high 5'-monodeiodinase and low 5-monodeiodinase activity was found. The 5'-monodeiodinase decreased during the first days of life whereas the 5-monodeiodinase activity remained at a low stable level until day 3 when the activities of both enzymes increased. The increase of conversion rate of T4 to T3 and rT3 in liver and kidney well correlate with the triiodothyronines concentration in serum from day 3 after birth.

Comparison of different methods for iodothyronines extraction from animal tissues.

Four different methods for thyroid hormones concentration in various tissues have been compared. The methods involve homogenization, extraction with 95% ethanol and assay of iodothyronines by a highly sensitive RIA. The method described by Chopra et al. has been selected and used for further estimations as the most reliable and convenient for a continuous laboratory work. The T4 levels (means +/- SE) found in the liver, kidney and skeletal muscle were 22.49 +/- 2.19, 14.89 +/- 1.56 and 8.33 +/- 1.73 ng per g wet tissue. The T3 contents were 8.53 +/- 1.08, 9.52 +/- 0.69 and 4.90 +/- 0.76 ng per g wet tissue, respectively.

Placental outer and inner ring monodeiodination of thyroxine and triiodothyronines in the rabbit.

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.