Effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin on guinea pig heart muscle

Cardiac effects of TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) were examined using atrial muscle isolated from young or mature guinea pigs fed ad libitum, from young animals 10 or 20 days after a single ip injection of 1 microgram TCDD/kg body wt, and from control guinea pigs pair-fed to the TCDD-treated animals. Basal contraction force of left atrial muscle obtained from TCDD-treated and pair-fed control guinea pigs 10 days after treatment was significantly increased compared to atrial muscle obtained from ad libitum control animals. Atrial muscle from TCDD-treated animals 20 days after treatment had a significantly lower basal force of contraction while that from pair-fed animals was significantly higher relative to atrial muscle from ad libitum control animals. These results suggest that the TCDD-induced change in basal atrial contraction force is in part due to undernutrition and that prolonged exposure to TCDD inhibits this response observed in pair-fed animals. Furthermore, prolonged TCDD treatment shifted the dose-response curve for the positive inotropic effect of isoproterenol to the right with no statistically significant change in the maximal inotropic effect obtained with high concentrations of isoproterenol. The TCDD-induced changes in force of contraction and the dose-response curve for the positive inotropic effect of isoproterenol were similar in young TCDD-treated and mature control guinea pigs. TCDD did not alter the basal contraction frequency or the dose-response curve for the positive chronotropic effect of isoproterenol. There was no effect on cardiac lipoprotein lipase activity at 1 microgram/kg, but in animals administered 4 micrograms TCDD/kg activity was reduced. It was concluded that TCDD adversely affects atrial muscle of guinea pig heart.     

Tissue-specific alterations of de novo fatty acid synthesis in 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-treated rats

De novo fatty acid synthesis was determined by the³H₂O method in numerous tissues and organs of TCDD-treated (125 g/kg), pair-fed and free-fed male Sprague-Dawley rats to investigate if this important pathway of intermediary metabolism is altered by TCDD. Of the 12 tissues and organs examined, liver showed an increased, and interscapular brown adipose tissue (IBAT) a decreased de novo fatty acid synthesis when comparing TCDD-treated to pair-fed or free-fed control rats. De novo fatty acid synthesis was unaffected in other organs and tissues examined, with the exception that the concentration of³H-fatty acids in plasma reflected the increased rate of synthesis seen in the liver of TCDD-treated animals. Increased de novo fatty acid synthesis in liver coincided with increased plasma triiodothyronine (T₃) concentrations, whereas decreased de novo fatty acid synthesis in IBAT parallelled decreased plasma thyroxine (T₄) levels. Thyroidectomy decreased de novo fatty acid synthesis, as expected, in both liver and IBAT. However, TCDD elicited no response in either of these organs in thyroidectomized rats. This finding suggests that changes observed in non-thyroidectomized rats are probably secondary effects. Indeed, known tissue-specific effects of T₃ on liver and T₄ on IBAT provide a likely explanation for the altered de novo fatty acid synthesis of these organs. It is suggested that increased de novo fatty acid synthesis in the liver of TCDD-treated rats might be responsible for the additional wasting away observable in these animals as compared to pair-fed controls.     

Relationship of the wasting syndrome to lethality in rats treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin

Young adult male Sprague-Dawley rats treated with a LD95 dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) exhibited a progressive reduction in feed intake and body weight until death occurred 15 to 32 days post-treatment. The time course and magnitude of weight loss and lethality of pair-fed control rats were essentially identical to that of TCDD-treated rats with each pair-fed control animal dying within 3 days of its TCDD-treated partner. Body composition analysis of the dead animals revealed that the total amounts of protein, fat, water, and ash in the carcasses of TCDD-treated and pair-fed control rats were each reduced to a similar extent. The temporal pattern of daily feed intake in TCDD-treated and pair-fed control rats (3 meals/day) or (1 meal/day) did not influence the results. Studies conducted at LD25-62 doses of TCDD in male Sprague-Dawley rats of different ages--weanling (90 g), young adult (275 g), and mature (450 g)--showed that the severity of the wasting syndrome in all age groups was greatest for animals that died. Also, young adult rats treated with a LD25 dose of TCDD that died displayed the same degree of hypophagia and weight loss prior to death as rats administered a LD95 dose. Histopathology of the liver and gastrointestinal tract was compared in TCDD-treated (LD95 dose) and pair-fed control rats killed 1 day before they otherwise would have died. Hepatocytes of TCDD-treated rats were enlarged relative to those of pair-fed control rats and contained nuclei that varied in size and number. Pair-fed control rats exhibited atrophy of the liver cords due to a decrease in the cytoplasmic volume of their hepatocytes. The stomach and small intestine of TCDD-treated rats were histologically similar to those of ad libitum-fed controls. In contrast, the glandular mucosa of the stomach of pair-fed control rats was ulcerated and the intestinal mucosa was atrophied. Stomach ulcers were the source of clotted blood found throughout the gastrointestinal tract of pair-fed control rats but not that of TCDD-treated animals. These findings demonstrate that hypophagia-induced weight loss is one of perhaps several responses that contribute to the death of TCDD-treated rats. That other responses are also involved is suggested by differences between pair-fed control and TCDD-treated rats in the weight and histopathology of certain organs. In addition, gastrointestinal blood loss contributes to the death of pair-fed control rats but not TCDD-treated animals.     

Hypophagia-Induced Weight Loss in Mice, Rats, and Guinea Pigs Treated with 2,3,7,8-Tetrachlorodibenzo-p-dioxin

Hypophagia-lnduced Weight Loss in Mice, Rats, and Guinea PigsTreated with 2,3,7,8-Tetrachlorodibenzo-p-dioxin. KELLING, C.K, CHRISTIAN, B. J., INHORN, S. L., AND PETERSON, R. E. (1985).Fundam. Appl. Toxicol. 5, 700–712. C57BL/6 mice treatedwith 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD; 360 µ/kg)displayed a significant reduction in feed intake and body weightuntil just before death, when they developed ascites and subcutaneousedema. This caused body weight of the mice that died to suddenlyincrease during the terminal stage of toxicity. TCDD-treatedmice that survived did not develop ascites or edema, and maintaineda body weight that was slightly less than that of pair-fed mice.Cumulative lethality in TCDD-treated mice (69%) was greaterthan that of pair-fed controls (14%). In guinea pigs treatedwith TCDD (2 µ/kg) both the time course and magnitudeof hypophagja were closely associated with weight loss. Pair-fedguinea pigs did not lose quite as much weight as TCDD-treatedanimals because their total body water content was higher. Waterintake in pair-fed guinea pigs was greater than that of TCDD-treatedanimals. The time course and magnitude of lethality tended tobe similar in TCDD-treated guinea pigs (81%) and pair-fed controls(64%). In Fischer F-344 rats treated with TCDD (100 µg/kg)body weight loss was associated with a reduction in both feedand water intake. The time course and magnitude of weight lossin TCDD-treated and pair-fed rats was essentially identical.Lethality was higher in TCDD-treated rats (95%) than pair-fedcontrol animals (48%). Taken together, these findings suggestthat hypophagia is responsible for the loss of adipose and leantissue in mice, guinea pigs, and rats treated with a LD70–95dose of TCDD. Under these dosage conditions, weight loss contributesmore to the lethality of guinea pigs than to that of FischerF-344 rats or C57BL/6 mice     

Comparative effects of pair-feeding and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on various biochemical parameters in female rats

Hypophagia is a common characteristic of the toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and may be responsible for some of the toxic manifestations. Pair-feeding has been used in control animals to compensate for the hypophagia, but relatively few studies have assessed biochemical changes associated with pair-feeding versus weight loss induced by TCDD. Rats were treated with TCDD and killed 7 days post-treatment while pair-fed animals received an amount of diet equivalent to TCDD-treated partner animals. Ad libitum-fed rats were also used. No correlations were seen in altered calcium and iron homeostasis between pair-feeding and TCDD administration relative to ad libitum-fed animals. Pair-feeding resulted in greater alterations than TCDD administration in the subcellular distribution of iron in mitochondria, microsomes and cytosol. Pair-feeding also resulted in greater accumulation of calcium in mitochondria and microsomes in pair-fed as compared to TCDD-treated animals. Greater lipid peroxidation was observed in whole liver and nuclei of rats receiving TCDD relative to pair-fed animals. A significantly greater incidence of DNA single strand breaks occurred in hepatic nuclei of TCDD-treated animals as compared to pair-fed and ad libitum-fed animals. Significantly greater inhibition of hepatic glutathione peroxidase activity and thymic involution were observed in TCDD treated animals as compared to the pair-fed group. Although some similarities existed between TCDD-treated animals and pair-fed rats, the overall biochemical changes which were observed following TCDD administration cannot be attributed to weight loss associated with hypophagia.     

Studies on the role of lipid peroxidation in the acute toxicity of TCDD in rats

Lipid peroxidation has been shown to be enhanced following exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), but its role in TCDD toxicity is unclear. The present study was undertaken to further elucidate the relations between lipid peroxidation and TCDD lethality. A time course and dose-response experiment in Long-Evans (L-E; LD50 ca. 10 micrograms/kg) and Han/Wistar (H/W; LD50 greater than 3000 micrograms/kg) rats showed that hepatic lipid peroxidation, measured as the amount of thiobarbituric acid-reactive substances (TBA-RS), was induced by TCDD dose-dependently in L-E, but not in H/W rats. Hepatic glutathione peroxidase activity was suppressed in much the same manner in both strains. Lipid peroxidation correlated with body weight loss in L-E rats alone. When 500 micrograms/kg of TCDD was given to L-E rats, lipid peroxidation increased about 3-fold on Day 11 in the liver, while no change was seen in cardiac or renal TBA-RS. The pair-fed controls did not survive the 11-day test period and exhibited gastrointestinal hemorrhages. At 6 days, liver atrophy and elevated (over 2-fold) TBA-RS values were recorded in pair-fed controls but not in their TCDD-treated counterparts. TCDD decreased hepatic glutathione peroxidase activity by almost 50% at 6 days, while pair-feeding was without effect. Liver morphology was different between TCDD-treated and pair-fed rats. Moreover, the livers of TCDD-treated L-E rats contained much higher concentrations of probably peripheral fat-derived fatty acids than did the livers of pair-fed or ad libitum control rats. Restricted feeding over 6 days induced hepatic lipid peroxidation more in H/W than in L-E rats. Endotoxin increased liver TBA levels similarly in both strains having an additive effect with high doses of TCDD in H/W rats. Added as a 0.5% concentration in chow, butylated hydroxyanisole (BHA), but not ethoxyquin, tended to increase survival rate and time in L-E rats exposed to 20 micrograms/kg of TCDD; at 50 micrograms/kg the only survivor was again in the BHA group. However, neither antioxidant had any effect on initial body weight loss. It is concluded that lipid peroxidation mainly arises as a secondary phenomenon in TCDD toxicity, is not the cause of the typical histopathological liver lesion, but may contribute to lethality.     

Intermediary metabolism of the mature rat following 2,3,7,8-tetrachlorodibenzo-p-dioxin treatment

Changes in body weight, feed intake, hepatic cellularity, and intermediary metabolism were assessed in the mature male (450 g) rat following 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) administration. All animals were schedule-fed (8-hr feeding period/24 hr) and treated with a single oral dose of either TCDD (75 micrograms/kg) or vehicle. Blood and tissues were sampled 16 to 18 hr following the end of the feeding period on 2, 4, 6, and 8 days post-treatment. Mature rats treated with TCDD exhibited a slight but progressive reduction in both body weight and feed intake throughout the 8-day experimental period. An increase in liver mass that was apparent at 2 days and plateaued by 4 days after TCDD treatment was associated with a decrease in the concentration of DNA per gram of wet liver. However, the total liver content of DNA in TCDD-treated rats remained similar to pair-fed animals. Thus, TCDD treatment produced liver enlargement in the mature rat that was the result of hepatocellular hypertrophy and not an increase in cell number. Hepatic glycogen content in TCDD-treated rats was threefold higher than their pair-fed counterparts at 2 to 6 days post-treatment, and this augmentation would account, in part, for the hypertrophy of the liver cell found after administration of TCDD. Plasma glucose and lactate concentrations were similar in TCDD-treated and pair-fed rats, suggesting that the Cori cycle remained unaltered following TCDD administration. Likewise, heart and gastrocnemius glycogen concentrations were similar in all experimental groups. Urinary excretion of urea, ammonia, and creatinine was comparable in TCDD-treated rats and their pair-fed counterparts, indicative of a nitrogen balance that was not disturbed by TCDD. Plasma glutamine concentrations in TCDD-treated rats tended to be reduced and were significantly lower at Day 6 post-treatment when compared to those of pair-fed counterparts, suggestive that amino acid release from muscle was not enhanced in TCDD-treated rats. Likewise, plasma concentrations of branched-chain amino acids, which are metabolized to a large extent in muscle, tended to be lower on Day 6 following TCDD treatment. Yet at Day 6 post-treatment, the circulating concentrations of amino acids that are metabolized by the liver were elevated in TCDD-treated animals. TCDD administration also resulted in an increase in total hepatic protein concentration which was evident at 4 days and increased progressively at 6 and 8 days post-treatment. Liver content of phospholipids also increased gradually following administration of TCDD.(ABSTRACT TRUNCATED AT 400 WORDS)     

Rabbit serum hypertriglyceridemia after administration of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)

TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) caused a dose-dependent decrease of adipose tissue lipoprotein lipase (LPL) activity and caused a concomitant increase in serum triglyceride concentration in the rabbit 10 d after single ip administration of either 1 or 50 micrograms/kg. Hepatic low-density lipoprotein (LDL) binding was markedly depressed and serum cholesterol concentrations were modestly increased relative to pair-fed control animals. Serum glucose concentrations were significantly lower in the rabbit administered TCDD compared to ad libitum or pair-fed control animals, although little change was observed in serum insulin concentration. Electron microscopic examination of aortic arches 20 d after a single ip administration of 50 micrograms TCDD/kg revealed ruffling, denudation, and sloughing off of the cell surface and the appearance of macrophage-like structures in the intima and media of the endothelial cells. These alterations resemble preatherosclerotic lesions typical in animals with hyperlipidemia. It is proposed that TCDD causes hyperlipidemia in the rabbit through suppression of LPL activity and LDL receptor binding.     

Studies on the Role of Lipid Peroxidation in the Acute Toxicity of TCDD in Rats*

Lipid peroxidation has been shown to be enhanced following exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), but its role in TCDD toxicity is unclear. The present study was undertaken to further elucidate the relations between lipid peroxidation and TCDD lethality. A time course and dose-response experiment in Long-Evans (L-E; LD50 ca. 10 μg/kg) and Han/Wistar (H/W; LD50 > 3000 μg/kg) rats showed that hepatic lipid peroxidation, measured as the amount of thiobarbituric acid-reactive substances (TBA-RS), was induced by TCDD dose-dependently in L-E, but not in H/W rats. Hepatic glutathione peroxidase activity was suppressed in much the same manner in both strains. Lipid peroxidation correlated with body weight loss in L-E rats alone. When 500 μg/kg of TCDD was given to L-E rats, lipid peroxidation increased about 3-fold on Day 11 in the liver, while no change was seen in cardiac or renal TBA-RS. The pair-fed controls did not survive the 11-day test period and exhibited gastrointestinal hemorrhages. At 6 days, liver atrophy and elevated (over 2-fold) TBA-RS values were recorded in pair-fed controls but not in their TCDD-treated counterparts. TCDD decreased hepatic glutathione peroxidase activity by almost 50% at 6 days, while pair-feeding was without effect. Liver morphology was different between TCDD-treated and pair-fed rats. Moreover, the livers of TCDD-treated L-E rats contained much higher concentrations of probably peripheral fat-derived fatty acids than did the livers of pair-fed or ad libitum control rats. Restricted feeding over 6 days induced hepatic lipid peroxidation more in H/W than in L-E rats. Endotoxin increased liver TBA levels similarly in both strains having an additive effect with high doses of TCDD in H/W rats. Added as a 0.5% concentration in chow, butylated hydroxyanisole (BHA), but not ethoxyquin, tended to increase survival rate and time in L-E rats exposed to 20 μg/kg of TCDD; at 50 μg/kg the only survivor was again in the BHA group. However, neither antioxidant had any effect on initial body weight loss. It is concluded that lipid peroxidation mainly arises as a secondary phenomenon in TCDD toxicity, is not the cause of the typical histopathological liver lesion, but may contribute to lethality.     

Alterations in splenocyte protein kinase C (PKC) activity by 2,3,7,8-tetrachlorodibenzo-p-dioxin in vivo

The effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on growth factor-coupled activation of nuclear protein kinase C (nPKC) and on the subcellular distribution of PKC activity in rat splenocytes were investigated. Seven days after a single injection of TCDD (50 μg/kg body weight), cytosolic and particulate PKC activity was significantly higher in splenocytes from TCDD-treated rats or pair-fed control rats compared to ad libitum-fed animals. In a separate experiment, purified splenocyte nuclei from TCDD-treated animals and controls were used to study activation of nPKC by growth factors and other trophic agents. Growth factor-stimulated nPKC activation was attenuated in splenic nuclei from TCDD-treated rats compared to vehicle-treated controls. Evidence presented here suggests that the cellular mechanism of TCDD toxicity leading to immunosuppression in rodents may be mediated in part by uncoupling of growth factor receptors linked to PKC activation at the level of the nucleus. However, changes in total splenocyte PKC activity appear to be correlated with hypophagia since cytosolic and particulate PKC levels were elevated in TCDD-treated rats and their pair-fed partners.     

Composition of diet modifies toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in cold-adapted rats

The effect of a high carbohydrate, high fat or high protein diet was studied on the acute toxicity of TCDD (125 g/kg) in cold-adapted (4±1° C) rats. Within 10 days after dosing, TCDD-treated rats fed a high carbohydrate or a high protein diet reduced their caloric intake by 25% whereas those fed a high fat diet consumed only 15% fewer kcal/MBS (metabolic body size). TCDD-treated rats fed a high protein diet lost body weight at the same rate as their pair-fed controls, whereas body weight loss in high fat-fed rats was significantly higher than in their pairfed controls. In contrast, TCDD-treated rats fed a high carbohydrate diet effectively maintained their body weight in the 4 days immediately after TCDD dosage, whereas their pair-fed controls lost weight. Mortality in TCDD-treated animals was 100% irrespective of the diet; all pairfed control rats (except one fed a high protein diet) were terminated on days corresponding to the spontaneous death of their TCDD-treated pairs. Mean time to 50% mortality and mean time to death were significantly longer in TCDD-treated rats fed a high carbohydrate diet in comparison with the other two TCDD-treated groups (p<0.05), although caloric intake was comparable. Serum triiodothyronine (T₃) was reduced in TCDD-treated animals fed a high fat or a high carbohydrate diet but not in those fed a high protein diet; serum thyroxine (T₄) was reduced in all the treated groups, irrespective of diet. The relative weight of liver was significantly higher in all TCDD-treated animals; the weight of pancreas was higher only in high carbohydrate-fed TCDD-treated rats and the weight of interscapular brown adipose tissue was lower only in high fat-fed TCDD-treated rats in comparison to pair-fed controls. It is concluded that TCDD-treated cold-adapted rats utilize carbohydrate and fat differently than pair-fed controls and that these changes probably contribute to the development of a wasting syndrome.This work was supported in part by a grant from the Bayerisches Staatsministerium für Landesentwicklung und Umweltfragen, München (Fed. Republic of Germany)     

Mode of metabolism is altered in 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-treated rats

Male Sprague-Dawley rats were fed either a high-fat (HF) or a high-carbohydrate (HC) diet and subsequently injected with either 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) (125 micrograms/kg) or vehicle (pair-fed controls). In all TCDD-treated animals, a reduction in caloric intake was evident as early as 1 day after dosage. Respiratory quotients (RQ) were determined at 5-day intervals. Their pattern for the HC-fed but not for the HF-fed TCDD-treated rats was different from that of the corresponding pair-fed controls. After an initial parallel decrease the RQ values remained low for TCDD-treated rats whereas they increased again for pair-fed controls. Serum total thyroxine (T4) was significantly lower in TCDD-treated animals and this reduction was not influenced by the composition of the diet. Serum triiodothyronine (T3) was neither altered by diet nor by TCDD. Thymic atrophy was as severe in pair-fed as in TCDD-treated rats fed the HC diet but not in rats fed the HF diet. Our results suggest that TCDD-treated rats are in a different mode of metabolism from pair-fed rats and that this difference is related to gluconeogenesis.     

Inhibition of intestinal glucose absorption in rats treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin.

Male Sprague-Dawley rats were injected with either 125 micrograms 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)/kg or vehicle (pair-fed and ad libitum-fed controls). Transfer of water, electrolytes and D-glucose as well as fats of a tracer dose of the non-metabolizable radioactive marker 3-O-methyl-D-[U-14C]glucose was studied in isolated perfused jejunal segments 1, 2, 7, and 21 d after treatment (TCDD-treated and pair-fed control rats) and after 26 d in ad libitum-fed controls. TCDD-treated rats demonstrated reduced feed consumption and loss of body weight. Active intestinal absorption of glucose was significantly inhibited 30 and 22% compared to pair-fed controls, respectively 2 and 7 d after TCDD treatment. After 21 d the inhibition (14%) was less significant. There were no differences in glucose transfer between severely starved pair-fed controls (body weights 370 +/- 26 g) and ad libitum-fed rats (body weights 512 +/- 15 g). Water absorption and transfer of sodium and calcium was not influenced by TCDD treatment. However, a significant increase of potassium transfer was observed in parallel with impaired glucose absorption. The uptake of 3-O-methylglucose into mucosal tissue was not impaired, whereas the transfer to the serosal side was significantly inhibited by 30-60% compared to pair-fed as well as ad libitum-fed animals from day 2 until the end of the experiment. These results suggest that TCDD is involved in an inhibition of glucose transport at the basolateral membrane.     

ROLE OF NON-ESTERIFIED FATTY ACIDS IN REGULATING PLASMA CHOLESTEROL TRANSPORT

1. The cholesteryl ester transfer protein (CETP) promotes an equimolar exchange of cholesteryl esters between the high density lipoproteins (HDL) and low density lipoproteins (LDL) in human plasma. 2. Sodium oleate converts this CETP-mediated process of exchange into one of net mass transfer of cholesteryl esters from HDL to LDL. 3. Thus, conditions which increase the concentration of non-esterified fatty acids in plasma may favour the redistribution of cholesterol from the non-atherogenic HDL to the atherogenic LDL fraction.     

Digestible energy and efficiency of feed utilization in rats treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin

Treatment of male rats (300 to 325 g) with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, 15 or 50 micrograms/kg) caused dose-dependent reductions in body weight, feed and water intakes, and fecal output. Urine output, however, was not altered by TCDD. Fecal energy loss, as a percentage of daily feed energy intake (kcal/day), was similar in control and TCDD-treated rats as was the percentage of feed energy absorbed by the gastrointestinal tract, i.e., digestible energy. These findings dispel the long-standing proposal that a gross malabsorption syndrome is responsible for weight loss in TCDD-treated rats and place greater emphasis on hypophagia as the reason for weight loss. In support of a central role for hypophagia, it was found that control rats pair-fed to rats treated with a sublethal dose of TCDD (15 micrograms/kg) lost almost the same amount of weight. However, from 15 to 50 days post-treatment, the pair-fed animals consistently maintained their weight at a 10- to 15-g higher level than age-matched TCDD-treated rats. To determine why this weight difference occurs, the efficiency of feed utilization from Day 30 to 45 post-treatment in ad libitum fed control and TCDD-treated rats (15 micrograms/kg) that were maintaining different levels of body weight was compared. First, daily feed intakes of TCDD-treated and control rats were determined from Day 25 to 30 post-treatment. Second, weights of both groups were lowered by reducing feed intake in two successive 5-day periods to 50 and 10% of the respective ad libitum level. Third, on Days 40 to 45, both groups were refed their prereduction level of intake but reduced in proportion to the intervening loss in metabolic tissue mass. At each level of feed energy reduction, weight losses observed in the TCDD-treated and control rats were equivalent. Furthermore, although given only prerestriction amounts of feed that were indexed to their reduced metabolic body size (body wt kg 0.75), both TCDD-treated and control rats gained weight rapidly and at similar rates during the refeeding period. Thus, rats treated with a sublethal dose of TCDD displayed normal efficiency of feed utilization but did so at a subnormal level of weight. That is, just like control rats, TCDD-treated rats increased their efficiency of feed utilization (weight gain/feed intake) but only when their body weight was caused to fall below the lower weight maintenance level determined by the TCDD dose administered.(ABSTRACT TRUNCATED AT 400 WORDS)     

Metabolism and disposition of 2,3,7,8-tetrachlorodibenzo-p-dioxin in guinea pigs

Marked interspecies variability exists in the acute toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), with the guinea pig being the mammalian species most sensitive to the acute toxicity of TCDD. The metabolism and disposition of TCDD was investigated in guinea pigs for 45 days following a single exposure to purified [3H]TCDD (0.56 microgram/kg, ip). Guinea pigs included in the toxicokinetic study gained body weight, maintained a normal relative body composition, and exhibited no gross signs of toxicity during the 45-day study. Approximately 36% of the dose of TCDD-derived 3H remained in the adipose tissue at 45 days following exposure to [3H]TCDD, while the liver, pelt, and skeletal muscle and carcass each contained about 7% of the administered dose. Although most of the TCDD-derived radioactivity in liver, kidney, perirenal adipose tissue, and skeletal muscle represented unchanged TCDD, from 4 to 28% of the 3H was associated with metabolites of TCDD. This unexpected finding suggests that TCDD metabolites are not efficiently excreted from guinea pigs. The urinary and fecal excretion of TCDD-derived radioactivity followed apparent first-order kinetics, with an elimination half-life of 93.7 +/- 15.5 days (mean +/- SD). HPLC analysis of urine and bile from [3H]TCDD-treated guinea pigs showed that all of the radioactivity represented metabolites of TCDD, indicating that these routes of elimination are dependent on prior metabolism of TCDD. However, 70 to 90% of the radioactivity in fecal samples was found to represent unmetabolized TCDD throughout the 45-day excretion study. The presence of TCDD in feces and its absence in bile suggest that the fecal excretion of unchanged TCDD resulted from the direct intestinal elimination of the lipophilic toxin. Furthermore, the cumulative excretion of TCDD-derived radioactivity over 45 days indicated that 74.3% of the 3H was excreted in feces as unchanged TCDD, while 25.7% of the 3H was excreted in urine and feces as TCDD metabolites. Thus, TCDD is primarily eliminated unchanged in the feces of guinea pigs, indicating that the metabolism of TCDD does not play a major role in the ultimate elimination of the toxin from the guinea pig. This may in part explain the relatively long excretion half-life for TCDD in the guinea pig and may contribute to the remarkable sensitivity of the guinea pig to the acute toxicity of TCDD.     

Effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on thermogenesis in brown adipose tissue of rats

Male Sprague-Dawley rats were treated with a usually lethal dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD; 125 micrograms/kg i.p. in corn oil) or with vehicle alone. Two, 4, and 8 days after treatment the temperature of interscapular brown adipose tissue (IBAT) was monitored during venous infusion of norepinephrine (480 ng/min) for 60 min. The temperature response was about 1.0-1.5 degrees C within 1 h in vehicle-treated, pair-fed and ad libitum-fed controls. In TCDD-treated animals, the response of IBAT decreased with time after TCDD dosage, amounting to only 0.3 +/- 0.1 degree C at 8 days after dosing (differences significant with respect to both controls, P less than 0.05). GDP binding to IBAT mitochondria (a measure of thermogenic capacity) was unchanged in all groups, indicating that the reduced thermogenic response was probably not caused by an impairment of the mitochondrial uncoupling process by TCDD.     

Transfer of 2,4,5,2',4',5'-hexachlorobiphenyl across the in situ perfused guinea pig placenta

The transplacental crossover of 14C-2,4,5,2',4',5'-hexachlorobiphenyl (6-CB) from the maternal circulation to the fetal side of the placenta was examined in intact fetuses and following the in situ perfusion of the guinea pig placenta. Fetal, late pregnant, and nonpregnant female guinea pig lipoprotein profiles and the association of 6-CB with these plasma constituents were also determined in vivo. Very low density lipoprotein (VLDL) concentrations were 10-fold higher in fetal than in maternal plasma, and the great majority of 6-CB which was transferred to intact fetuses became associated with this plasma fraction. 6-CB was found primarily in association with low density lipoproteins (LDL) in nonpregnant animals. In the late pregnant guinea pig, 6-CB became primarily associated with plasma protein in spite of circulating protein concentrations lower than those seen in the nonpregnant state. No differences in the levels of the three plasma lipoprotein classes were observed between pregnant and nonpregnant animals. It was found that an amount of 6-CB similar to that found in intact litter mates crossed the perfused placenta over the same time period. Despite the much higher VLDL concentrations on the fetal side of the placenta and the association of 6-CB with VLDL in intact fetuses, addition of 1,000 mg/dl VLDL to the 5.4% bovine serum albumin perfusion medium failed to influence the magnitude of 6-CB crossover. 6-CB crossover was influenced by protein concentration in the perfusion media in a concentration-dependent fashion. It is hypothesized that 6-CB and free fatty acids traverse the placenta and are retained by the fetus via similar mechanisms.     

Relationship of Alterations in Energy Metabolism to Hypophagia in Rats Treated with 2,3,7,8-Tetrachlorodibenzo-p-dioxin

Relationship of Alterations in Energy Metabolism to Hypophagiain Rats Treated with 2,3,7,8-Tetrachlorodibenzo-p-dioxin. POTTER,C. L., MENAHAN, L. A., AND PETERSON, R. E. (1986). Fundam. Appl.Toxicol. 6, 89–97. Efficiency of energy utilization wasevaluated temporally in 2,3,7,8-tetrachlorodibenzo-p-dioxin(TCDD, 50 µg/kg)-treated male Sprague—Dawley rats(275–300 g), their pair-fed counterparts, and a groupwith ad libitum access to ground feed. TCDD treated rats exhibiteda progressive reduction in feed intake and body weight. Theweight loss of vehicle-treated rats, pair-fed to the TCDD-treatedgroup, was comparable to that found in rats receiving TCDD.Following treatment, rats administered TCDD were as efficientin absorbing feed energy from the gut as control rats. Thiswas evidenced by similar relative digestible energy values inTCDD-treated rats, their pair-fed partners, and a group withad libitum access to feed. Equivalent decreases in oxygen consumptionand carbon dioxide production in TCDD-treated rats and theirpair-fed counterparts, relative to rats with ad libitum accessto feed, suggested that the decrease in both of these parametersin TCDD-treated rats was secondary to hypophagia and/or weightloss. Decline of respiratory quotient (RQ) to almost 0.7 inboth TCDD-treated rats and their pair-fed counterparts is indicativeof fat combustion. By Day 17 post-treatment, RQ increased significantlyin the TCDD-treated and pair-fed groups possibly due to a limitationin the availability of lipid stores. Also, TCDD-treated ratsand their pair-fed partners diminished their water intake toa similar extent without reducing urine output. Likewise, urinaryexcretion of both energy and urea was decreased to the sameextent in rats treated with TCDD as it was in their pair-fedcounterparts. However, TCDD-treated rats tended to excrete moreurinary ammonia than their pair-fed partners on Days 10 and16 post-treatment. Thus, TCDD treatment does result in a reductionof caloric intake in the rat, yet efficiency of energy utilizationis preserved.     

Increased copper concentrations in rat tissues after acute intoxication with 2,3,7,8-tetrachlorodibenzo-p-dioxin

Recently, acutely toxic doses of the environmental pollutant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) have been reported to affect the hepatic distribution of essential metals in the rat. However, the reduced food intake by TCDD was not taken into account. Therefore, metal concentrations were determined in different rat tissues at the end of a toxicity study with TCDD in which a pair-fed control group was introduced. Male Sprague-Dawley ats received a single i.p. injection of corn oil/acetone with or without TCDD at 125 g/kg. Controls and TCDD-treated rats were fed ad libitum; additionally, pair-fed controls received the amount of food consumed by their TCDD-treated partners 1 day before. Twenty-one days after dosing rats were killed and samples of liver, kidney and jejunum were taken for the analysis of Ca, Cu, Fe, Mg, Mn, and Zn. After acid digestion of the tissues metals were determined by atomic emission spectrometry (AES). The most outstanding effect of TCDD treatment was an increase of the copper levels in the kidney (4-fold, versus pair-fed controls) and in the liver (>2-fold, versus pair-fed controls). Other metals were mainly affected by the reduced food intake only. Since Cu represents a trace metal the homeostasis of which depends on its biliary excretion and since TCDD is known to impair biliary flow and excretion, an impaired biliary excretion of Cu by TCDD is suggested as the causal mechanism.Supported by the Bayerisches Staatsministeriuni für Landesentwicklung und Umweltfragen