Relative roles of the thyroid hormones and noradrenaline on the thermogenic activity of brown adipose tissue in the rat.

We have assessed the relative contribution of the thyroid hormones and noradrenaline (NA) on the calorigenic function of brown adipose tissue (BAT) as indicated by GDP binding and O2 consumption of BAT mitochondria. Male Wistar rats of 200 g body weight were made hypothyroid with 131I. Groups of animals were injected s.c., in divided doses, daily for 10 days, with thyroxine (2 micrograms/100 g body weight) or tri-iodothyronine (T3; 0.3 microgram/100 g body weight). Animals were used 7 days after bilateral or unilateral sympathetic nerve excision of BAT (Sx). Sham-operated rats were used as controls. In normal rats kept at 22 degrees C, GDP binding reached 94 +/- 24 pmol/mg protein; untreated hypothyroid rats had normal binding values whereas the T3-treated group showed an increased binding. Sx induced a sharp fall in the three groups (P < 0.01). After 24-h exposure to 4 degrees C GDP binding increased in normal rats to about 410% (P < 0.01) whereas binding failed to increase in response to cold in the untreated hypothyroid and the T3-treated groups. Sx reduced GDP binding in the three groups significantly (P < 0.01). The consumption of O2 by BAT mitochondria showed similar variations in response to Sx and to cold exposure as did GDP binding. The data indicated that, at room temperature, BAT calorigenesis can function without the thyroid hormones, though not without the catecholamines. The findings in rats exposed to cold showed that the lack of NA was significantly more effective than the lack of thyroid hormones in preventing the BAT hyperactive response. This does not negate an active role for T3 in BAT calorigenesis.

Epidermal growth factor stimulates thyrotropin secretion in the rat.

The present work studied the effects of epidermal growth factor (EGF) on the release of thyrotropin (TSH) and prolactin (PRL) from perifused pituitary glands of 200-gram male Wistar rats. Each pituitary gland, cut into halves, was placed in a chamber of a perifusion system connected to a peristaltic pump which conveyed the perifusion medium (Medium 199, pH 7.3, Gibco, USA) from a reservoir to a chamber at a flow rate of 100 microliters/min. Each tightly closed chamber contained one pituitary gland and 600 microliters medium and it was placed in a water bath at 37 degrees C throughout the experiment. One milliliter samples of effluent were collected every 10 min for 60 min to obtain baseline values of TSH and PRL. Thereafter, TSH-releasing hormone (TRH) 10(-8) M or EGF (10(-11), 10(-10), 10(-9) or 10(-8) M) were added to individual chambers and the 10-min sampling of effluent continued for 60 min. EGF 10(-11) M elicited no TSH response, but 10(-10) and 10(-9) M doses induced significant increases in TSH secretion (p < 0.01) with a peak at 10 min after addition of EGF. In another experiment, EGF 10(-8) M or TRH 10(-8) M significantly elevated TSH secretion (p < 0.01). However, TRH, but not EGF, stimulated PRL secretion (p < 0.01). In the in vivo studies, the intravenous administration of EGF 10(-5) M or TRH 10(-5) M both induced significant elevation of TSH release at 10 min after the injection (p < 0.02 for EGF and p < 0.01 for TRH).(ABSTRACT TRUNCATED AT 250 WORDS)

Diphenylhydantoin stimulates the intrapituitary conversion of thyroxine to triiodothyronine in the rat.

Treatment of normal rats with diphenylhydantoin (DPH) decreases serum thyroxine (T4) and triiodothyronine (T3) levels without the anticipated rise in serum thyrotropin (TSH). The present work has studied the intrapituitary conversion of T4 to T3 in male Wistar rats, 200-250 g body weight (BW), treated with DPH 5 mg/100 g BW/day for 8 days. A tracer dose of 3',5'-[125I]T4 (150 microCi) was injected intravenously, and 2 h later hypophyses were removed and homogenized individually at 4 degrees C in ice-cold PBS buffer (pH 7.4). T4 and T3 were extracted in 400 microliters n-butanol:2 N HCl (9:1) and chromatographed in tertiary amyl alcohol:hexane: 1 N ammonia (5:1:6). In 11 untreated control rats, [125I]T3 generated from [125I]T4 deiodination was 35 +/- 6% and intact [125I]T4 was 49 +/- 9% of total chromatographic radioactivity. In 11 DPH-treated rats [125I]T3 increased (p < 0.001) and [125I]T4 decreased (p < 0.02). The DPH effect was blocked in rats treated for 2 days with iopanoic acid 10 mg/100 g BW, though blocking was not seen in rats treated with half the dose of iopanoic acid. In normal rats receiving supplemental doses of T4 (2 micrograms/100 g BW/day for 8 days), DPH similarly increased pituitary 5'-deiodination. Administration of propylthiouracil (PTU) to T4-supplemented rats had no effect on pituitary 5'-deiodination of T4, whereas the addition of DPH to PTU treatment increased [125I]T3 production (p < 0.01). Serum T4 (p < 0.001) and T3 (p < 0.01) were decreased after DPH therapy, while serum and pituitary TSH were not altered.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of thyroid hormones on mitochondrial oxygen consumption in brown adipose tissue and heart from cold-exposed hypothyroid rats.

The present work measured brown adipose tissue and heart mitochondrial oxygen consumption in hypothyroid rats treated with replacement doses of T3, T4 or T4 plus iopanoic acid and kept at 4 degrees C for 24 h. Heart oxygen consumption in normal, untreated hypothyroid and T4-treated hypothyroid rats was unaffected by cold exposure. In rats treated with T4 plus iopanoic acid, rates of oxygen consumption were normal in those maintained at 4 degrees C as well as in those kept at room temperature, despite serum T3 concentration being significantly decreased. The cold-exposed T3-treated hypothyroid rats showed a marked decrease in oxygen consumption (p less than 0.02) and alpha-glycerophosphate dehydrogenase activity, a T3-dependent enzyme. Mitochondrial oxygen consumption in brown fat from normal (p less than 0.01), T4 (p less than 0.02) and T4 plus iopanoic acid-treated (p less than 0.01) rats rose more than twofold in response to cold. In the T3-treated group, oxygen consumption at room temperature was higher (p less than 0.02) than in any other group at similar temperatures. However, the T3-treated group showed no changes in oxygen consumption in response to cold, perhaps because this group reached the maximal response at room temperature. The untreated and the T3-treated hypothyroid rats (both groups devoid of T4) did not survive at 4 degrees C unless T4 or several-fold replacement amounts of T3 were administered. The data demonstrate the crucial role of T4 in thermogenesis during cold exposure.

[Effects of cold on myocardial mitochondria respiration of rats with hypothyroidism treated with triiodothyronine]. / Efectos del frío sobre la respiración mitocondrial del miocardio de ratas hipotiroideas tratadas con triiodotironina.

The present work studied the effect of cold on oxygen consumption (OC) and alpha-glycerophosphate dehydrogenase activity (alpha-GPD) in heart mitochondria of hypothyroid rats (hypo) treated with T3, T4 or T4 plus Iopanoic Acid (IOP). 200 g male Wistar rats were made hypothyroid by 131I administration. Animals were injected s.c., in divided doses, for 10 days, with one of the following substances: T3, 300 ng/100 g BW/day; T4, 2 micrograms/100 g BW/day or T4 plus IOP, 5 mg/100 g BW/day, for 72 h preceding the experiment. One half of each group was housed in a cold room at 4 degrees C and the other at 22 degrees C, for 25 h, and thereafter decapitated. Heart mitochondria were isolated by routine methods. The OC was measured polarographically using L-malate, L-glutamate and malonate as substrates. Intramitochondrial alpha-GPD activity was measured by a microcolorimetric assay. The results from 16 or 20 rats/group (4 or 5 pools of 4 hearts each) were: In the rats kept at 22 degrees C the OC (in ng at. oxyg./min/mg prot.; State 3) in the hypo+T4 group was 69 +/- 10; in the rats treated with T4+IOP, 75 +/- 11 and in the hypo+T3, 102 +/- 5. When the animals were exposed to 4 degrees C no change was observed in the hypo+T4 and hypo+T4IOP groups. On the other hand, OC was significantly lower in the T3-treated animals (p less than 0.001, versus their controls at 22 degrees C). This group of rats did not survive when exposed to cold.(ABSTRACT TRUNCATED AT 250 WORDS)

[The effect of epidermal growth factor on thyrotropin secretion in rats]. / Acción del factor de crecimiento epidérmico sobre la secreción de tirotrofina en la rata.

The present work studied the effects of epidermal growth factor (EGF) on the secretion of thyrotropin (TSH) from perifused pituitaries of 200 g body weight male Wistar rats. After decapitation the neural lobe was discarded and the anterior pituitary was transferred to a chamber of a perifusion system connected to a peristaltic pump which conveyed the perifusion medium (Medium 199) through a reservoir to a chamber at a flow rate of 100 microliters/min. Individual chambers were filled with 600 microliters of medium and placed in a water bath at 37 degrees C. One ml samples of effluent were collected every 10 min for 60 min to obtain baseline values of TSH. Thereafter, TSH-releasing hormone (TRH) (10(-8) M) or EGF in varied concentrations (10(-8) M to 10(-11) M) were added to individual chambers. The 10 min sampling of effluent was then continued for 60 min to measure TSH by RIA (NIADDK, rTSH RP-2 standard). In the TRH study, the mean basal TSH concentration was 32.1 +/- 6.5 ng/ml, increasing to 105 +/- 13.8 ng at 10 min post-TRH (P less than 0.005) and declining to basal values at 20 min. Addition of EGF 10(-8) M increased TSH secretion from a mean basal value of 68.9 +/- 5.6 ng/ml to 201 +/- 44.3 ng/ml (P less than 0.02) and a return to normal value at 20 min. Similar effects were induced by EGF 10(-9) M (P less than 0.001) and 10(-10) M (P less than 0.05) whereas no effect was elicited by EGF 10(-11) M.(ABSTRACT TRUNCATED AT 250 WORDS)

Efectos del frío sobre la respiración mitocondrial del miocardio de ratas hipotiroideas tratadas con triiodotironina. / [Effects of cold on myocardial mitochondria respiration of rats with hypothyroidism treated with triiodothyronine]

The present work studied the effect of cold on oxygen consumption (OC) and alpha-glycerophosphate dehydrogenase activity (alpha-GPD) in heart mitochondria of hypothyroid rats (hypo) treated with T3, T4 or T4 plus Iopanoic Acid (IOP). 200 g male Wistar rats were made hypothyroid by 131I administration. Animals were injected s.c., in divided doses, for 10 days, with one of the following substances: T3, 300 ng/100 g BW/day; T4, 2 micrograms/100 g BW/day or T4 plus IOP, 5 mg/100 g BW/day, for 72 h preceding the experiment. One half of each group was housed in a cold room at 4 degrees C and the other at 22 degrees C, for 25 h, and thereafter decapitated. Heart mitochondria were isolated by routine methods. The OC was measured polarographically using L-malate, L-glutamate and malonate as substrates. Intramitochondrial alpha-GPD activity was measured by a microcolorimetric assay. The results from 16 or 20 rats/group (4 or 5 pools of 4 hearts each) were: In the rats kept at 22 degrees C the OC (in ng at. oxyg./min/mg prot.; State 3) in the hypo+T4 group was 69 +/- 10; in the rats treated with T4+IOP, 75 +/- 11 and in the hypo+T3, 102 +/- 5. When the animals were exposed to 4 degrees C no change was observed in the hypo+T4 and hypo+T4IOP groups. On the other hand, OC was significantly lower in the T3-treated animals (p less than 0.001, versus their controls at 22 degrees C). This group of rats did not survive when exposed to cold.(ABSTRACT TRUNCATED AT 250 WORDS)

Acción del factor de crecimiento epidérmico sobre la secreción de tirotrofina en la rata. / [The effect of epidermal growth factor on thyrotropin secretion in rats]

The present work studied the effects of epidermal growth factor (EGF) on the secretion of thyrotropin (TSH) from perifused pituitaries of 200 g body weight male Wistar rats. After decapitation the neural lobe was discarded and the anterior pituitary was transferred to a chamber of a perifusion system connected to a peristaltic pump which conveyed the perifusion medium (Medium 199) through a reservoir to a chamber at a flow rate of 100 microliters/min. Individual chambers were filled with 600 microliters of medium and placed in a water bath at 37 degrees C. One ml samples of effluent were collected every 10 min for 60 min to obtain baseline values of TSH. Thereafter, TSH-releasing hormone (TRH) (10(-8) M) or EGF in varied concentrations (10(-8) M to 10(-11) M) were added to individual chambers. The 10 min sampling of effluent was then continued for 60 min to measure TSH by RIA (NIADDK, rTSH RP-2 standard). In the TRH study, the mean basal TSH concentration was 32.1 +/- 6.5 ng/ml, increasing to 105 +/- 13.8 ng at 10 min post-TRH (P less than 0.005) and declining to basal values at 20 min. Addition of EGF 10(-8) M increased TSH secretion from a mean basal value of 68.9 +/- 5.6 ng/ml to 201 +/- 44.3 ng/ml (P less than 0.02) and a return to normal value at 20 min. Similar effects were induced by EGF 10(-9) M (P less than 0.001) and 10(-10) M (P less than 0.05) whereas no effect was elicited by EGF 10(-11) M.(ABSTRACT TRUNCATED AT 250 WORDS)

Acción del factor de crecimiento epidérmico sobre la secreción de tirotrofina en la rata / The effect of epidermal growth factor on thyrotropin secretion in rats

El factor de crescimiento epidérmico (EGF) es un polipéptido de potente acción mitogénica. La probable influencia de esto factor sobre la fisiología del eje hipofiso-tiroideo no ha sido establecida. En el presente trabajo se estudió la acción del EGF sobre la secreción de TSH por hipófisis de ratas Wistar macho in vitro. Cada adenohipófisis fue colocada en una cámara de purifusión conectada a una bomba peristáltica que impulsaba el fluído de perifusión (medio 199, pH 7,3) a un ritmo de 100 *l/min. Cada cámara, conteniendo una hipófisis y 600 *l de fluído de perifusión, fue mantenida en un baño a 37§C. Se obtuvieron muestras del effluente (1 ml cada 10 min.) durante 60 min. para medir TSH basal. Luego se agregó en cámaras individuales, TRH en concentración final de 10-8 M o EGF en concentraciones de 10-8 M a 10-11 M, luego de lo cual se continuócon la colección del efluente cada 10 min. por otros 60 minutos. En cada muestra se midió TSH (ng/ml) por RIA (NIADDK, rTSH-RP 2 standard). Resultados: en el estudio con TRH, la TSH basal promedió 32,1 ñ 6,5 ng/ml con un pico post-TRH de 105 ñ 13,8 ng/ml a los 10 min. (P<0,005), retornando al valor basal ...

Acción del factor de crecimiento epidérmico sobre la secreción de tirotrofina en la rata / The effect of epidermal growth factor on thyrotropin secretion in rats

El factor de crescimiento epidérmico (EGF) es un polipéptido de potente acción mitogénica. La probable influencia de esto factor sobre la fisiología del eje hipofiso-tiroideo no ha sido establecida. En el presente trabajo se estudió la acción del EGF sobre la secreción de TSH por hipófisis de ratas Wistar macho in vitro. Cada adenohipófisis fue colocada en una cámara de purifusión conectada a una bomba peristáltica que impulsaba el fluído de perifusión (medio 199, pH 7,3) a un ritmo de 100 *l/min. Cada cámara, conteniendo una hipófisis y 600 *l de fluído de perifusión, fue mantenida en un baño a 37ºC. Se obtuvieron muestras del effluente (1 ml cada 10 min.) durante 60 min. para medir TSH basal. Luego se agregó en cámaras individuales, TRH en concentración final de 10-8 M o EGF en concentraciones de 10-8 M a 10-11 M, luego de lo cual se continuócon la colección del efluente cada 10 min. por otros 60 minutos. En cada muestra se midió TSH (ng/ml) por RIA (NIADDK, rTSH-RP 2 standard). Resultados: en el estudio con TRH, la TSH basal promedió 32,1 ñ 6,5 ng/ml con un pico post-TRH de 105 ñ 13,8 ng/ml a los 10 min. (P<0,005), retornando al valor basal ... (AU)