P. de Martino Rosaroll

The direct effect of the thyroid hormone on cardiac chronotropism.

To establish whether thyroid hormone modifies the heart rate directly or through an action on other neuroendocrine modulators, the authors have examined several animals models differing in the plasma levels of such compounds. Induction of the hypothyroid state in rats produced a slow onset of bradycardia, which may be removed by a prolonged triiodothyronine treatment. The involvement of TSH was excluded as, by comparing thyroidectomized, hypophysectomized and cold exposed rats, the heart rate was found to vary according to the thyroid levels and not to the TSH levels. Moreover growth hormone, corticotropin and gonadotropins do not influence the heart rate, as the bradycardia induced by hypophysectomy was fully removed by triiodothyronine treatment. The lack of influence by ACTH and GnH was confirmed by treatment of thyroidectomized rats with corticosteroids or testosterone, respectively. Finally, thyroid hormone did not act on the heart rate by changing the norepinephrine output at the sympathetic nerve endings in the heart. In fact, thyroidectomy produced a more intense bradycardia than sympathectomy, and such bradycardia was equally removed by triiodothyronine treatment in thyroidectomized rats and in thyroidectomized and then sympathectomized ones. The authors suggest that the direct effect of the thyroid hormone on cardiac chronotropism is due to an early enhancement of beta-adrenoceptors, followed by a late modification of the electrophysiological properties of the myocardium.

Electrophysiological properties of papillary muscle fibres from euthyroid and hypothyroid chick.

1. PTU treatment-induced hypothyroidism is associated with a significant decrease in the chick heart rate. 2. Hypothyroidism produces a slow onset of bradycardia, indicating a late effect of thyroid hormone decrease. 3. After 15 days treatment, an increase of action potential duration, similar to that reported for several mammal species, has been found. 4. Action potential duration is frequency and temperature dependent, but it has been found to be significantly different in the euthyroid and hypothyroid chicks.

Electrophysiological properties of the hyperthyroid rat heart

We have studied the effects of in vivo administration of different T3 doses to thyroidectomized rats on electrophysiological properties, measured in vitro, of papillary muscle fibers. The treatment with increasing T3 doses was associated with a significant reduction of the action potential duration up to a dose as large as 25 micrograms/100 g body weight every second day. The treatment with larger doses of T3 tended to restore the values of the action potential duration present in animals treated with physiological doses (5 micrograms/100 g body weight every second day). Action potential duration is frequency dependent. As the stimulation rate was increased from 1 to 5 Hz, this duration increased in all groups. However the difference between the rat groups remained significant. The cardiac frequency measured in unanaesthetized rats increased as the T3 doses. Furthermore the intrinsic frequency showed a similar increase, indicating a direct effect of T3 on the pacemaker cells in all thyroid states. The mechanism of this action of the thyroid hormone is not, however clear.

Effect of Thyroid State on the Morphofunctional Properties of Heart Mitochondria

For an evaluation of the effect of thyroid hormone on the mitochondrial compartment of heart, we have studied morphological and functional characteristics of mitochondria within the heart of rats in different thyroid states. In particular, the relationship between membrane-bound proteins, such as the cytochromes, and the surface of inner mitochondrial membranes has been investigated. By quantitative electron microscopy it has been shown, according to our previous data, that thyroidectomy causes an increase in several parameters such as the specific mitochondrial mass (mg mitochondrial protein/g tissue), the number and inner membrane surface of mitochondria per unit volume of heart, without any significant variation of the ratio between the surface of the inner membranes and the volume of mitochondria. The treatment with substitutive doses of triiodothyronine (T3) tends to restore the normal pattern. On the other hand, mitochondria isolated from thyroidectomized rats show lower oxygen consumption due to a lower content of cytochromes, which are distributed at lower density on the inner membranes. Also in this case the pattern is reversed by T3 treatment. It is concluded that thyroid hormone causes an increase of mitochondrial cytochrome content, which, notwithstanding the decrease of the mitochondrial mass, leads to a higher respiratory rate and to a larger capacity of oxidative metabolism in the heart.

Effect of T3 administration on electrophysiological properties of lizard ventricular muscle fibres.

We investigated the effects on the electrophysiological properties of ventricular muscle fibres from lizards kept at 20 °C of mild and severe hyperthyroidism. The hyperthyroidism was induced by a 4-day treatment with either 0.025 or 1.0 μg triiodothyronine g-1 body weight, documented by increased serum levels of thyroid hormone. Triiodothyronine treatment did not modify the duration of the action potential recorded in vitro at 25 °C from ventricular muscles stimulated at 1 Hz. Recordings at higher temperatures were associated with a faster repolarization phase and a decrease of action potential duration in both euthyroid and hyperthyroid animals. However, in lizards treated with 1.0 μg triiodothyronine · g-1 body weight, the 90% repolarization recovery times at 30 and 35 °C (95.6±14.9 ms and 53.0±6.0 ms, respectively), were significantly shorter than normal (177.6±29.2 and 107.2±18.1 ms, respectively). Action potential duration was also dependent on stimulation frequency of the preparations. Increased frequency led to significant decrease of the duration of action potentials recorded at 25 °C. In euthyroid preparations the reductions in 90% repolarization recovery time, owing to increases in stimulation frequency to 2.5 and 5 Hz, were 19.3±1.7 and 35.6±2.0 ms, respectively. In hyperthyroid preparations, the reductions in the 90% recovery time due to stimulus frequency increases varied from 35.4±1.9 and 58.1±2.1 ms at low hormone doses to 38.9±2.0 and 58.2±2.1 ms at high hormone doses. As a result of these differences, the action potential durations recorded from the two hyperthyroid preparations at high stimulation rates were shorter than from euthyroid preparations. The results obtained suggest that lizard cardiac tissue is responsive to hormone action at low environmental temperature, but the effects of such action become evident when the temperature and heart rate increase.

Effect of cold exposure on electrophysiological properties of rat heart

Male rats exposed to the cold (4°C) for five or ten days exhibited modifications in their thyroid state, as documented by increases in serum thyroid hormone levels, to which differently graded modifications of heart weight/body weight ratio, heart rate, and resting metabolic rate were associated. The values of the above mentioned thyroid state indicators returned to those of the control when the animals, kept at cold for ten days, were re-exposed to room temperature (24°C) for an additional 10 days. The configuration of action potentials, recorded in vitro at 26°C from fibres of anterior papillary muscles, was different in control rats of different age and was affected by prolonged cold exposure. In fact, the action potential duration (APD) increased after ten days of cold exposure. In the re-exposed group the APD was not different from that of the controls. Such a pattern was not significantly modified when the stimulation frequency increased from 1 Hz to 5 Hz. The above results suggest that in cold exposure, as in experimental hyperthyroidism, thyroid hormone might exert a cardiac chronotropic effect by modifying heart electrophysiological properties. Thus thyroid hormone should play a basic role during the exposure to cold environment, stimulating the body metabolism and increasing heart rate as a response to the requirement for greater tissue perfusion.

Modification of electrophysiological properties of rat heart with age

The authors have determined the serum thyroid hormone levels [total (TT3) and free (FT3) triiodothyronine], the heart weight/body weight ratio and the heart rate of differently aged male rats. The variations of these parameters show a modification of thyroid state as a function of ageing. The authors have also recorded, at about 26 degrees C, resting and action potentials from single cells of papillary muscles isolated from the same groups of rats. The animals in the higher thyroid state exhibited a repolarization speed higher than the other animals. The thyroidectomy, performed on 50 day old rats, and T3 treatment of the thyroidectomized rats give rise to modifications of repolarization speed and then of action potential duration analogous to ones obtained in previous study for animals thyroidectomized at 30 days of age. These data demonstrate that the modifications of heart electrophysiological properties with age, are due fundamentally to thyroid state modifications. The results suggest also that the cardiac chronotropism modifications which the rat undergoes as a function of ageing are due to the changes of levels of thyroid hormone which might exert its effect by modifying the ion channel kinetics as well as the cardiac receptors.

Ventricular electrophysiological properties in normal and congenitally hypothyroid neonatal rats

The serum thyroid hormone levels [total (TT3) and free (FT3) triiodothyronine] and the heart rates were determined in neonatal rats of different ages (1-5-10 days). Thyroid hormone levels increase gradually in the first 10 days of age. The heart rate, tested at a body temperature of 37 degrees C, also increases during the same period. As the increase in heart rate in this phase of rat life is not due to the catecholamines, it is suggested that such an increase might depend on the increased thyroid hormone activity. On the other hand in congenitally hypothyroid rats the levels of both hormones and heart rates are lower than in normal animals of the same age. The electrophysiological properties of ventricular muscle fibres include a longer action potential, irrespective of stimulation frequency, in younger, naturally hypothyroid animals. The duration of action potential is greater in the congenitally hypothyroid animals, at all ages. These data demonstrate that, as in young and adult rats, the age-related modifications in heart rate, found in neonatal rats, might be due to thyroid dependent modifications of cardiac electrophysiology.

Modulation of the heart rate by thyroid hormone in the newborn rat

Confirming the literature data the authors describe that the heart rate is smaller in the newborn rats than in adult ones and increases until the adult values during the first two weeks of life. On the other hand, the blood thyroid hormone exhibits the same pattern, showing an early postnatal increment. As, according the Adolphs data (1967), the heart rate enhancement is not due to the catecholamines, the authors suppose that such enhancement might conceivably depend on thyroid hormone increment.

Effect of Thyroid Hormone on the Mitochondrial DNA and RNA Levels in Rat Cardiocytes

Normal rats show levels of heart mitochondrial DNA and RNA (mit-DNA, mit-RNA) which are 0.66 and 18.82 µg/mg of mitochondrial proteins, or 0.15 and 4.35 fg/mitochondrion, respectively. Thyroidectomy reduces such values, while triiodothyronine (T3) treatment effectively restores normal levels of mitochondrial nucleic acids. These findings complete the pattern obtained previously by the authors for the mitochondrial nucleic acids of liver and suggest that T3 also increases the heart mitochondrial DNA and RNA content, probably due to stimulation of the respective mitochondrial polymerases.