Seven weeks feeding of magnesium and fluoride modifies plasma lipids of hypercholesterolaemic rats in late growth phase.

Recent findings indicated that a low fluoride supplement, especially with a low magnesium supplement in the basically low magnesium diet of genetically hypercholesterolaemic male RICO rats, may prevent the generation of atherosclerotic serum lipid profile. In the present study, several plasma lipids/lipoproteins were measured in the same strain of rats after a later growth phase. The control group C was fed an adequate diet with 45 per cent sucrose plus some cholesterol while the dietary fluoride was very low (1.1 mg F/kg of diet). In diet of group D, the Mg content was reduced to about one seventh of Mg of group C, i.e. to 122 mg/kg. Diet of group E was as that of group D with F content elevated to 17.2 mg/kg. Diet of group G was as that of group E with Mg content elevated to 220 mg/kg. The feeding period was terminated at 12 h deprivation of food and following exsanquination. Total plasma cholesterol in group C was 4.5 mmol/L. The central factor in causing reduction in growth rate and several plasma lipids and their lipoprotein subfractions appeared to be the magnesium deficiency. However, the greatest significant reductions in plasma cholesterol, plasma free cholesterol, plasma and VLDL esterified cholesterol and also HDL cholesterol, HDL esterified cholesterol and plasma triglycerides from those levels of the control group C were found in group G (fluoride and magnesium supplements). Supplementation of fluoride alone in group E reduced only plasma and VLDL esterified cholesterol. In the present male RICO rats, low dietary F and Mg supplements, separately, and especially together, may participate in the regulation of the outcome of atherosclerosis via affecting several plasma lipid risk factors known to associate with the development of atherosclerosis.

Changes in thyroid hormone and thyrotrophin in the serum and thyroid glands of developing genetically obese male and female Zucker rats.

In a first experiment, serum thyroxine (T4), 3,5,3'-triiodothyronine (T3) and thyrotrophin (TSH) concentrations as well as thyroid gland T4 and T3 contents were measured in developing lean and obese Zucker male and female rats of 4-16 weeks of age. The rats were bred in our laboratory and always treated in sex-matched pairs of one lean and one obese rat from the same litter. Serum T4 was not different in any phenotype/sex group at 4 weeks. In male rats, it became progressively lower (27 and 37% at 12 and 16 weeks respectively) in obese than in lean rats. In females, similar levels of serum T4 were maintained in both obese and lean developing rats. Serum T3 was similar in obese and lean male 4-week-old rats whereas it was lower (28%) in obese than in lean females. It became progressively lower (39 and 49% at 12 and 16 weeks respectively) in obese than in lean developing male rats. In females, lower levels of serum T3 were maintained (25 and 43% at 12 and 16 weeks respectively) in obese than in lean rats. Serum TSH was not different in any phenotype/sex group at 4 weeks. It rose in both obese and lean male rats with age, but became progressively lower (33 and 23% at 12 and 16 weeks respectively) in obese compared with lean rats. In females, similar levels of serum TSH were maintained in both obese and lean developing rats. Thyroid gland weight was not different in any phenotype/sex group at 4 weeks.(ABSTRACT TRUNCATED AT 250 WORDS)

Marked increases of two kinds of two-exon-skipped albumin mRNAs with aging and their further increase by treatment with 3'-methyl-4-dimethylaminoazobenzene in Nagase analbuminemic rats.

Nagase analbuminemic rats (NARs) have a 7-base-pair deletion at the 5' splice site of the HI intron of the albumin gene. The level of immunohistochemically albumin-positive hepatocytes is about 1 per 10(5) cells in neonatal NARs, increases with age, and further increases with chronic oral treatment with 3'-methyl-4-dimethylaminoazobenzene (3'-MeDAB). The mechanisms involved in the increase in albumin-positive hepatocytes during aging of NARs and their treatment with 3'-MeDAB were analyzed. NARs were found to have four species of albumin mRNA: intact mRNA and those lacking the regions corresponding to exon H, exon G-H, and exon H-I. In 4-week-old NARs, the level of intact albumin mRNA was about 1/4000 of that in normal rats and mRNA lacking the exon H sequence was the major species. In aged and 3'-MeDAB-treated aged NARs, all four species of mRNA increased and the relative proportion of mRNAs lacking two exon sequences to mRNAs lacking one exon sequence was greatly increased, suggesting that aging is associated with changes of the splicing pattern and that 3'-MeDAB treatment enhanced these changes. In aged NARs and 3'-MeDAB-treated aged NARs, there was an increase in the amount of aberrant 60-kDa albumin. The 60-kDa protein could be a translation product of mRNAs lacking two exons, the amount of which increases in aged NARs and 3'-MeDAB-treated NARs.

The spontaneously epileptic rat (SER), a zitter*tremor double mutant rat: histopathological findings in the central nervous system.

The spontaneously epileptic rat (SER), a mutant homozygous for both zitter and tremor genes, exhibits absence-like seizures and tonic convulsions without external stimulation from 7 to 8 weeks of age. Histopathological studies of the central nervous system revealed the following abnormalities. The 35-day-old SERs which exhibit body tremor, and which have never shown seizures, had marked vacuolation and hypomyelination in the brainstem and cerebellum. The vacuoles were produced by splitting of the myelin sheaths and swelling of the dendrites and were related to primary swelling of the astrocytes. The 2- to 3-month-old SERs with staggering gait and seizures showed focal axonal swelling ('torpedo') and advanced vacuolation in the granular cell layer of the cerebellum in addition to the abnormalities observed at 35 days of age. Degenerative neurons and spheroidal bodies were observed in the substantia nigra and ventral tegmental nucleus. These brain areas are known to be related to tonic convulsions in the several experimental models. The SER is believed to be a useful tool for the investigation of the relationship between the structure and function of the central nervous system in epilepsy. It is probable that the more severe changes in the cerebellum are responsible for the staggering gait.

Growth responses in a mutant dwarf rat to human growth hormone and recombinant human insulin-like growth factor I.

A new mutant GH-deficient dwarf rat has been used to study the effects of iv infusions of human GH (hGH) and recombinant human insulin-like growth factor I (hIGF-I). This animal has only about 5% of normal pituitary GH content, low circulating GH levels, and no regular GH surges. The defect seems to be specific for GH. Infusions of hIGF-I at 180 micrograms/day for 9 days elevated serum IGF-I concentrations significantly over those in the saline-infused controls (713 +/- 20 ng/ml vs. 395 +/- 31 ng/ml); hGH infusions did not raise IGF-I levels significantly (435 +/- 20 ng/ml). Gel filtration of serum samples showed that the high-dose hIGF-I infusions increased free IGF concentrations, without apparently altering the pattern of IGF-I binding whereas hGH infusions increased the amount of high mol wt IGF-I binding protein. Neither IGF-I nor hGH infusions affected the small amounts of rat GH present in the dwarf rat pituitary glands. Continuous iv infusions of hGH (200 mU/day for 9 days) stimulated body wt gain (2.1 +/- 0.2 g/day) and bone growth (96 +/- 9 microns/day) significantly compared to saline-infused dwarf rats (1.2 +/- 0.3 g/day and 43 +/- 3 microns/day). Infusions of hIGF-I at 180 micrograms/day produced a body wt gain (2.1 +/- 0.5 g/day) similar to that seen in the hGH-infused group but a significantly smaller stimulation of bone growth (63 +/- 3 microns/day). Infusion of a 5-fold lower dose of hIGF-I (36 micrograms/day for 9 days) had no effect on body wt or bone growth. Food intake was unaffected by either hGH or hIGF-I infusions. The pattern of tissue growth was affected differentially by hGH and IGF-I infusions that produced the same overall body wt gain. hGH induced a relatively proportional growth in most of the organs studied, whereas hIGF-I infusion at 180 micrograms/day stimulated a disproportionately greater growth of the kidney, adrenals, and spleen. In some of the animals, tissues were extracted for RIA of IGF-I; the amounts of IGF-I in the liver were similar in control, hGH, or IGF-I-infused animals, whereas kidney and adrenals from IGF-I infused animals contained larger amounts of immunoreactive IGF-I than did those tissues from hGH-treated rats. Thus, both hGH and hIGF-I can promote growth in the mutant dwarf rat, but they differ both quantitatively and qualitatively in their pattern of actions.

Sexual dimorphism of hyperglycemia and glucose tolerance in Wistar fatty rats.

Obese and lean male and female Wistar fatty rats were fed a high-sucrose (68% of calories) diet from 5 to 22 wk of age. Obese males, but not obese females, developed hyperglycemia in the fed state and were more glucose intolerant during an intragastric glucose tolerance test than obese females. Lean Wistar fatty rats did not become hyperglycemic on the sucrose diet. Obese males also showed a smaller insulin response during the glucose tolerance test than did obese females. The Wistar fatty rat is a sexually dimorphic model of non-insulin-dependent diabetes mellitus in which the male but not the female obese rats become diabetic. The diabetic condition and impaired glucose tolerance in the obese male Wistar fatty rat may be related to impaired pancreatic insulin release and peripheral insulin resistance.

Insulin-dependent changes in subcellular distribution of liver insulin receptors in obese Zucker rats.

Acute hyperinsulinaemias induced by insulin and stimulants of insulin secretion have been shown to cause a translocation of liver insulin receptors from the cell surface to the intracellular compartment, with little or no change in total receptor number. To determine whether a similar phenomenon occurs in chronic hyperinsulinaemic states, we have carried out a longitudinal study of total, cell surface and intracellular liver insulin receptors in genetically obese Zucker rats, with spontaneously develop hyperinsulinaemia. Liver plasma membranes, Golgi-endosomal fractions, a microsomal fraction and a total particulate fraction were isolated in 2-14-week old obese (fa/fa) rats and examined for specific insulin binding relative to lean (Fa/?) age-matched animals. In 16-day old rats, which were still normoinsulinaemic, insulin binding was unchanged. Later on, as hyperinsulinaemia developed, three sequential changes in insulin binding activity were observed: first, a 25-30% increase in Golgi-endosomal fractions (20 days); then, a 50-60% decrease in Golgi-endosomal fractions (4-5 weeks); and finally, a 50% decrease in plasma membranes (11 weeks), microsomal fraction and total particulate fraction (14 weeks), accompanied by restoration in Golgi-endosomal fractions (8-11 weeks). Unlike insulin receptors, insulin extractable from Golgi-endosomal fractions at 4-5 weeks was unchanged or increased. We conclude that, although an early increase in the endocytosis of liver insulin receptors may occur in hyperinsulinaemic Zucker rats, this mechanism does not account for the later decrease in cell surface receptors observed in these animals.

Evidence of hypothyroidism in the genetically epilepsy-prone rat.

A number of neurochemical and behavioral similarities exist between the genetically epilepsy-prone (GEPR) rat and rats made hypothyroid at birth. These similarities include lower brain monoamine levels, audiogenic seizure susceptibility and lowered electroconvulsive shock seizure threshold. Given these similarities, thyroid hormone status was examined in GEPR rats. Serum samples were collected from GEPR-9 and non-epileptic control rats at 5, 9, 13, 16, 22, 31, 45, 60, 90, 150 and 350 days of age. Serum thyroxine (T4) levels were significantly lower in GEPR-9 rats compared to control until day 22 of age. GEPR-9 thyrotropin (TSH) levels were significantly elevated during the period of diminished serum T4. GEPR-9 triiodothyronine (T3) levels were lower than control throughout the first year of life. The data indicate that the GEPR-9 rat is hypothyroid from at least the second week of life up to 1 year of age. The critical impact of neonatal hypothyroidism on brain function coupled with the development of the audiogenic seizure susceptible trait by the GEPR-9 rat during the third week after birth suggests that neonatal hypothyroidism could be one etiological factor in the development of the seizure-prone state of GEPR-9 rats.

Early development of the thyroid axis in the Brattleboro rat.

Plasma concentrations of thyrotrophin (TSH), thyroxine (T4), and triiodothyronine (T3), and pituitary TSH concentrations were determined at weekly intervals during the first 42 days following birth in Brattleboro homozygous (DI), Brattleboro heterozygous (HZ), and Long-Evans (LE) rats. Offspring from matings of Brattleboro rats were divided into DI and HZ animal subgroups on the basis of hypothalamic vasopressin content. In control LE rats, circulating levels of TSH, T4, and T3, and pituitary TSH concentrations increased during the early postnatal period to reach relatively stable levels between 28 and 42 days of age. In DI and HZ rats, the thyroid axis developed in parallel to that of LE rats during initial postnatal weeks. However, by 42 days of age, pituitary TSH concentrations were clearly elevated in Brattleboro rats relative to levels in age-matched LE animals. These data indicate that differences in thyroid axis function between Brattleboro and LE rats occur only after the attainment of a degree of maturity.

Immunocytochemical localization of retinoid-binding proteins in developing normal and RCS rats.

Interstitial retinol-binding protein (IRBP), cellular retinol-binding protein (CRBP), and cellular retinal-binding protein (CRALBP) were localized in developing normal and RCS rat retinas. IRBP is found in the presumptive interphotoreceptor space in normal and RCS rats on P1. It reaches adult levels on P18. In the RCS rat, the level of IRBP decreases after P18 until there is only a small amount of staining on P38. CRBP and CRALBP are localized in the retinal pigment epithelium from P1 to adult. CRALBP is also found in the ciliary body pigment epithelium and in the outer epithelium of the iris during the first week, but decreases during the second week to a very low level as found in the adult. The significance of the early presence of IRBP and the transient appearance of CRALBP in the ciliary body and iris epithelia is discussed.

Skeletal muscle growth in lean and obese Zucker rats.

Obese Zucker rats exhibit marked hyperphagia when compared to lean littermates but deposit a smaller percentage of total dietary energy as body protein. This study was designed to determine the roles of skeletal muscle protein synthesis, protein degradation, RNA, or DNA in producing the lower muscle mass of obese rats. At 44 days, 3 hindlimb muscles, the extensor digitorum longus (EDL), the gastrocnemius and the plantaris were significantly smaller in the obese animals. At 72 days, the differences in weights of these muscles were more pronounced. Protein synthesis and degradation were determined in the soleus at 44 days of age using an in vitro whole muscle incubation technique. Protein synthesis rate was significantly decreased in the obese animals. These changes were accompanied by reductions in both RNA and DNA levels. Significant changes in nucleic acid levels were observed in both the red and white portions of the gastrocnemius muscle. These changes in the anabolic process of protein accretion appear to be sufficient to account for the reduced muscle mass in the obese Zucker rat.

Growth and survival of RCS dystrophic rats fed modifications of the AIN-76 diet.

The RCS dystrophic rat is a hooded, pigmented-eyed strain widely used as a model for retinal degeneration. In addition to progressive photoreceptor loss, this strain suffers from unexplainable high mortality during the suckling period when reared on commercial cereal-based diets. Supplementation of these diets with 25% sunflower seeds greatly reduces this problem. This report presents the results of a study undertaken to test the effectiveness of the purified AIN-76 diet in controlling the high mortality. Growth of F1 rats and percent survival of the F2 offspring were determined for RCS dystrophic rats provided one of 5 diets: (Control) modified AIN-76 diet (15% additional sucrose in place of starch, nd vitamin and mineral mixes made up in cellulose rather than powdered sucrose); control with double the content of vitamin mix; control with double the content of fat; control with double the content of mineral mix; and control with double the content of protein. The percent survival of the F2 offspring was increased from 73% to 92% by vitamin supplementation. Doubling the mineral or protein content proved lethal to 54% and 79% of offspring on these respective diets. Increasing the vitamin content also resulted in an improvement in the initial growth of females. Increased fat, mineral or protein content all decreased the growth rate of males. Likewise, growth of females was decreased when the mineral and protein content of the diet was raised.