Pigmentation and dysfunction of Gunn rat thyroid

The thyroid gland of homozygous Gunn rats is moderately enlarged and displays a brownish-black discoloration. Light microscopic examination discloses that the follicular cells are filled with brown granules, which are shown, under the electron microscope, to be modified colloid droplets. Most of them possess a strong acid phosphatase and a mild peroxidase activity and contain a melanin-like pigment, according to histochemical analysis. In comparison with normal Wistar rats, Gunn rats possess significantly higher plasma thyroxine and lower triiodothyronine as well as an increased plasma TSH level. The soluble protein content of the thyroid is reduced in the Gunn rat, as is the total intrathyroid iodine content. The hyperthyroxinaemia of homozygous Gunn rats is due to a hereditary deficiency in hepatic glucuronyl transferase activity. The excess circulating thyroxine is of little functional importance because it is firmly bound to plasma proteins. But Gunn rats have a slight hypothyroid goitre for reasons not yet elucidated. The functional as well as morphological data at present available suggest a modified thyroid iodine metabolism and an altered composition of the thyroglobulin which may induce abnormalities in colloid proteolysis. The observed pigment may result from peroxidation of tyrosine. These alterations are probably independent of the sole enzymatic deficiency so far encountered in these animals and may probably be ascribed to a primary enzymatic defect in the thyroid gland itself.     

Radioactive iodoprotein in thyroid lymph and blood

1. Samples of thyroid and non-thyroid lymph and of thyroid and peripheral venous blood were obtained from primates and cats that had previously been given radioactive iodine. The distribution of the organic radioiodine between the protein and non-protein fractions in these samples was determined. 2. The proportion of the organic radioiodine in the form of iodoprotein was assessed by paper chromatography, acid-ethanol precipitation, hot-butanol washing, column chromatography and separate estimation of iodotyrosines after enzymic hydrolysis. 3. In thyroid lymph the relative proportion of the organic radioiodine in the form of iodoprotein was 75–98%; in blood it was much lower, probably no more than 6–7%. The absolute concentration of iodoprotein radioactivity also was considerably greater in thyroid lymph than in blood. 4. Enzymic hydrolysis of the protein of the thyroid lymph yielded a pattern of iodoamino acids that corresponded closely with that obtained after hydrolysis of protein extracted from the thyroid gland itself. 5. It can be concluded that the iodoprotein in thyroid lymph consists mainly of thyroglobulin or a closely related compound.     

A correlation between protein thermostability and resistance to proteolysis

The loss of activity due to proteolysis of purified L-asparaginase and beta-galactosidase from different sources correlates with the thermal instability of the enzymes. A similar correlation is found when populations of soluble proteins from micro-organisms grown at different temperatures are compared for proteolytic susceptibility and thermal stability. It is proposed that there is a general correlation between the thermostability of proteins and their resistance to proteolysis.     

Context-dependent resistance to proteolysis of intrinsically disordered proteins

Intrinsically disordered proteins (IDPs), also known as intrinsically unstructured proteins (IUPs), lack a well-defined 3D structure in vitro and, in some cases, also in vivo. Here, we discuss the question of proteolytic sensitivity of IDPs, with a view to better explaining their in vivo characteristics. After an initial assessment of the status of IDPs in vivo, we briefly survey the intracellular proteolytic systems. Subsequently, we discuss the evidence for IDPs being inherently sensitive to proteolysis. Such sensitivity would not, however, result in enhanced degradation if the protease-sensitive sites were sequestered. Accordingly, IDP access to and degradation by the proteasome, the major proteolytic complex within eukaryotic cells, are discussed in detail. The emerging picture appears to be that IDPs are inherently sensitive to proteasomal degradation along the lines of the "degradation by default" model. However, available data sets of intracellular protein half-lives suggest that intrinsic disorder does not imply a significantly shorter half-life. We assess the power of available systemic half-life measurements, but also discuss possible mechanisms that could protect IDPs from intracellular degradation. Finally, we discuss the relevance of the proteolytic sensitivity of IDPs to their function and evolution.     

Study of distribution and interaction of arsenic and selenium in rat thyroid

Seventy-eight Wistar weanling rats were pretreated with arsenate (100 mg/L As), selenite (1 mg/L Se), and arsenate (100 mg/L As) plus selenite (1 mg/L Se) added to the drinking water. After 4 w, all the animals were sacrificed and serum T3 and T4 were determined by double-antibody radioimmunoassay. Thyroid tissue concentrations of As and Se were determined in female rats by neutron activation analysis, and tissue specimens were examined histopathologically. For both sexes, the measurements indicated that T4/T3 was lowest in the Se group, intermediate in the As group, and highest in the controls. Corrected for the mean value of the controls, mean As concentration of thyroid tissue was of the same magnitude in the group pretreated with As+Se as the sum of the mean As concentration in the groups pretreated with As or Se alone. The outcome was symmetric with regard to the Se concentration: In the As+Se pretreated group, the mean Se concentration was of the same magnitude as the sum of the mean Se concentration in the groups pretreated with As or Se alone. Thus, As and Se tended to accumulate in the thyroid tissue. Postmortem examination showed that the thyroid tissue of rats pretreated with As alone exhibited obvious, toxic changes, whereas only minor or no changes were found in the tissues of the groups pretreated with Se or As+Se, and in the tissues of the controls. Multivariate analyses demonstrated that s-T4 and s-T3 were significantly correlated with sex, that s-T3 was positively correlated (p≤0.001) with Se pretreatment, and that the T4/T3 ratio was negatively correlated with both As (p≤0.012) and Se pretreatment (p≤0.001). The results were discussed in relation to the cancer preventive effect of Se.     

Calcitonin cell population and distribution in the thyroid gland of the rat

Calcitonin-containing cells (C cells) were identified in male Wistar white rats using an immunoperoxidase technique. They occupied a central position within the thyroid; very few were found peripherally, inferiorly, and superiorly; and none were present in the isthmus. The number of calcitonin-containing cells present per gram of body weight increased with age up to 70 days and had declined by 100 days. Determining the true total C-cell count through the entire thyroid is a very laborious procedure. However, a simple estimate of this total count can be made; the total number of C cells in every tenth section (6 microns) of thyroid was found to be highly correlated with the weight of the animal expressed as an allometric function. A better estimate can be derived from counts of just three sections: the tenth, twentieth, and thirtieth after the section of greatest cross sectional area.     

ATP-dependent proteolysis contributes to the acclimation-induced drought resistance in spring wheat

The effect of water deficit on the ATP-dependent proteolysis and total protein degradation was estimated in the leaves of spring wheat (Triticum aestivum L.) acclimated and non-acclimated to drought. The rate of ATP-dependent proteolysis, quantified as a difference between degradation of ¹²⁵I-lysozyme under ATP-regenerating and ATP-depleting systems, accounted for about 55 % of total ¹²⁵I-lysozyme degradation in fully turgid wheat leaves. In the non-acclimated leaves dehydration decreased sharply ATP-dependent proteolysis catalyzed by proteasome down to about 5% while in the leaves acclimated to drought water deficit raised ATP-dependent proteolysis to 87 % of total ¹²⁵I-lysozyme hydrolysis.     

Pituitary resistance to thyroid hormones.

Pituitary thyroid hormone resistance (PRTH) refers to a particular form of thyroid hormone refractoriness that is accompanied by peripheral hyperthyroidism, as only the TSH-secreting pituitary cells appear to be resistant to the effects of thyroid hormones. The presence of PRTH is suspected and diagnosed on the basis of the finding of high free thyroid hormone levels along with unsuppressed TSH, clinical signs and symptoms of hyperthyroidism and values of at least one of the parameters evaluating peripheral thyroid hormone action in the hyperthyroid range. However, most patients with PRTH present with clinical signs and symptoms of thyroid dysfunction, particularly goiter and tachycardia, overlapping those recorded in patients with generalized thyroid hormone resistance (GRTH), i.e. refractoriness to thyroid hormones at both pituitary and peripheral tissue level. Moreover, most of them display normal values of other parameters evaluating the peripheral effects of thyroid hormones and bear mutations in the gene encoding for T3 nuclear receptors similar to those found in patients with GRTH. These findings are questioning the existence of PRTH as a separate clinical entity and support the view that the various forms of thyroid hormone resistance may be part of a spectrum of disease with variable expression in different issues.     

Transfer of the heritable nonhemolytic jaundice trait from the Gunn rat to the Sprague-Dawley rat

Congenital nonhemolytic jaundice as observed in the Gunn rat was transferred successfully to the Sprague-Dawley rat. This jaundice trait occurs as the result of a deficiency of bilirubin glucuronyltransferase and appeared to transfer by simple Mendelian inheritance. A comparison of jaundiced Gunn with jaundiced Gunn-Sprague-Dawley cross rats for plasma bilirubin level, bilirubin glucuronyltransferase activity and female reproductive performance showed no significant difference between the two jaundiced rat groups. The phenotypic expression of the jaundice trait as viewed by the parameters used in this study appeared to be the same for both the Gunn and Gunn-Sprague-Dawley cross rats. The transfer of the jaundice trait to another rat strain enhances the opportunity to characterize this animal model and to determine the possible influence of a long-term closed mating system.     

Heterogeneity of collagen molecules types I and II according to their resistance to proteolysis

Study of the effects of pepsin treatment on soluble collagens type I of the skin and collagens type II of the costal cartilage of healthy subjects revealed the presence of two classes of molecules differing in the stability of their three-helical structure. In collagen molecules possessing a low stability (their number may amount to 20-30%) within the temperature range of 4-30 degrees C pepsin causes a split-off of N-terminal sites with the formation of short chains, i.e., alpha 1(I), alpha 2(II), and alpha 1(II), whereas at higher temperatures (33 degrees C for collagens type I and 37 degrees C for collagens type II) a complete degradation of these molecules takes place. It was found that collagens types I and II molecules contain a high number of three-helical sites with a high susceptibility to pepsin. The putative functional role of structural heterogeneity of collagen molecules is discussed.     

The effect of iodide administration on hog thyroid gland and the composition of thyroglobulin and 27-S iodoprotein.

The effect of excess iodide on hog thyroid gland has been examined with regard to the change in the chemical composition of thyroglobulin and in the accumulation of 27-S iodoprotein by the in vivo treatment of hogs with iodide for various lengths of time. The iodine content of thyroglobulin was either unchanged by short term administration of excess iodide, or somewhat lowered. However, the iodine content as well as the total amount of thyroglobulin increased in the glands enlarged by prolonged treatment with iodide. The iodine highest reached 1.17% of the protein on an average. On the other hand, 27-S iodoprotein decreased and finally disappeared after the chronic treatment. Monoiodotyrosine and diiodotyrosine increased in parallel with the increase in the iodine content (0.15 to 1.17%) caused by the iodide treatment, while thyroxine increased but reached a plateau at the level of three residues per mole of thyroglobulin, and no change was observed even in the proteins with the higher iodine content than 0.75%. Proteolytic activity measured by amino acid release from the thyroid protein was depressed by the chronic treatment. On the other hand, the amount of iodocompound released by the autoproteolysis, which may reflect hormone secretion, increased, possibly because of the marked increase in the iodine content of thyroglobulin.     

Methamphetamine-induced spectrin proteolysis in the rat striatum

Methamphetamine (METH) is a widely abused psychostimulant. Multiple high doses of METH cause long-term toxicity to dopamine (DA) and serotonin (5-HT) nerve terminals in the brain, as evidenced by decreases in DA and 5-HT content, decreases in tyrosine and tryptophan hydroxylase activities, decreases in DA and 5-HT re-uptake sites, and nerve terminal degeneration. Multiple high doses of METH are known to elicit a rapid increase in DA release and hyperthermia. Although METH also produces a delayed and sustained rise in glutamate, no studies have shown whether METH produces structural evidence of excitotoxicity in striatum, or identified the receptors that mediate this toxicity directly, independent of alterations in METH-induced hyperthermia. These experiments investigated whether METH can cause excitotoxicity as evidenced by cytoskeletal protein breakdown in a glutamate receptor-dependent manner. METH increased calpain-mediated spectrin proteolysis in the rat striatum 5 and 7 days after METH administration without affecting caspase 3-dependent spectrin breakdown. This effect was completely blocked with the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor antagonist, GYKI 52466, but not the NMDA receptor antagonist, MK-801. However, AMPA or NMDA receptor antagonism did not attenuate the METH-induced depletions of the dopamine transporter (DAT). Independent mechanisms involved in mediating spectrin proteolysis and DAT protein loss are discussed.     

Control of proteolysis in perifused rat hepatocytes

The mechanism by means of which amino acids inhibit intrahepatic protein degradation has been studied in perifused rat hepatocytes. Proteolysis was extremely sensitive to inhibition by low concentrations of amino acids. A mixture of 0.5 mM leucine and 1-2 mM alanine, concentrations found in the portal vein of the rat after feeding, inhibited proteolysis to the same extent as a complete physiological mixture of amino acids. Inhibition by these two amino acids was accompanied by a rise in the intracellular concentrations of glutamate and aspartate, and was largely prevented by addition of glucagon, by addition of the transaminase inhibitor aminooxyacetate, or by omission of K+. Acceleration of proteolysis by K+ depletion was accompanied by a fall in intracellular glutamate caused by an increased rate of transport of this amino acid to the extracellular fluid. It is concluded that intracellular leucine, glutamate and aspartate are important elements in the control of hepatic protein degradation.     

Inhibiting mitochondrial-dependent proteolysis of Mcl-1 promotes resistance to DNA damage

Elimination of Myeloid Leukemia Cell 1 (Mcl-1) is an early event in the onset of cell death following DNA damage and in many settings plays a critical role in dictating the success of chemotherapeutic agents. Following DNA damage, Mcl-1 is rapidly and efficiently targeted to the 26S proteasome through the action of E3 ubiquitin ligases. Tumors having acquired lesions that lead to stabilization of Mcl-1 are highly aggressive and have a poorer prognosis. Herein, we further characterize an additional mechanism of Mcl-1 proteolysis that is proteasome-independent but mitochondrial-dependent. A mitochondrial targeting signal located in the N-terminus of Mcl-1 is essential for targeting Mcl-1 to this alternative degradative avenue. We demonstrate that the Akt/mTORC1 survival pathway protects Mcl-1 from mitochondrial-dependent proteolysis. Disrupting Mcl-1 mitochondrial targeting improves the pro-survival capacity of Mcl-1 both ex vivo and in vivo in the well characterized mouse Eμ-Myc lymphoma model. Our data uncover an important relationship between the mitochondria and the Mcl-1 N-terminus in dictating cell fate following DNA damage.     

Immunochemical and immunohistochemical studies on the 27 S iodoprotein of dog thyroid with reference to thyroglobulin-like reaction of the parafollicular cells.

Our earlier finding that the thyroglobulin-like material responsible for the immunoreaction of parafollicular cells obtained in peak I fraction of Bio-Gel A-5m was followed up in the present study by an investigation of the immunochemical and immunohistochemical reactions of 27 S iodoprotein which was the most prominent material in the peak I fraction. The antibody was raised against completely purified 27 S iodoprotein which was obtained as follows: Thyroglobulin was extracted from dog thyroids and chromatographed initially on Bio-Gel A-5m and then on Bio-Gel A-50m. The area of 27 S migrated as a single bank on polyacrylamide gel slab electrophoresis. This was cut and eluted. Anti-27 S antiserum showed the same immunochemical patterns to 27 S and 19 S as anti-19 S antiserum with three different immunochemical methods: double diffusion test, one dimensional and two dimensional immunoelectrophoresis. The immunoperoxidase reactions of the anti-27 S antiserum and anti-19 S antiserum were restricted to follicular cells and luminal colloids. No reaction of the parafollicular cells was obtained by these antisera. Thus, 27 S iodoprotein shared common immunochemical and immunohistochemical properties with 19 S thyroglobulin. It was concluded that 27 S iodoprotein was not responsible for the thyroglobulin-like reaction of the parafollicular cells.