Restricted feeding induces daily expression of clock genes and Pai-1 mRNA in the heart of Clock mutant mice

Plasminogen activator inhibitor-1 (PAI-1) is a key factor of fibrinolytic activity. The activity and mRNA abundance show a daily rhythm. To elucidate the mechanism of daily Pai-1 gene expression, the expression of Pai-1 and several clock genes was examined in the heart of homozygous Clock mutant (Clock/Clock) mice. Damping of the daily oscillation of Pai-1 gene expression in Clock/Clock mice was accompanied with damped or attenuated oscillations of mPer1, mPer2, mBmal1, and mNpas2 mRNA. Daily restricted feeding induced a daily mRNA rhythm of all clock genes and Pai-1 mRNA in Clock/Clock mice as well as wild-type mice. The peaks of clock genes and Pai-1 mRNA were phase-advanced in the heart of both genotypes after 6 days of restricted feeding. The present results demonstrate that daily Pai-1 gene expression depends on clock gene expression in the heart and that a functional Clock gene is not required for restricted feeding-induced resetting of the peripheral clock.     

Maternal obesogenic diet enhances cholestatic liver disease in offspring

Human and animal model data show that maternal obesity promotes nonalcoholic fatty liver disease in offspring and alters bile acid (BA) homeostasis. Here we investigated whether offspring exposed to maternal obesogenic diets exhibited greater cholestatic injury. We fed female C57Bl6 mice conventional chow (CON) or high fat/high sucrose (HF/HS) diet and then bred them with lean males. Offspring were fed 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) for 2 weeks to induce cholestasis, and a subgroup was then fed CON for an additional 10 days. Additionally, to evaluate the role of the gut microbiome, we fed antibiotic-treated mice cecal contents from CON or HF/HS offspring, followed by DDC for 2 weeks. We found that HF/HS offspring fed DDC exhibited increased fine branching of the bile duct (ductular reaction) and fibrosis but did not differ in BA pool size or intrahepatic BA profile compared to offspring of mice fed CON. We also found that after 10 days recovery, HF/HS offspring exhibited sustained ductular reaction and periportal fibrosis, while lesions in CON offspring were resolved. In addition, cecal microbiome transplant from HF/HS offspring donors worsened ductular reaction, inflammation, and fibrosis in mice fed DDC. Finally, transfer of the microbiome from HF/HS offspring replicated the cholestatic liver injury phenotype. Taken together, we conclude that maternal HF/HS diet predisposes offspring to increased cholestatic injury after DDC feeding and delays recovery after returning to CON diets. These findings highlight the impact of maternal obesogenic diet on hepatobiliary injury and repair pathways during experimental cholestasis.     

Impaired daily glucocorticoid rhythm in Per1 Brd mice

Biological clocks have evolved in all kinds of organisms in order to anticipate and adjust to the daily light–dark cycle. Within the last decade, the molecular machinery underlying the circadian system was unraveled. In the present study, the impact of the loss of the Per1 or Per2 genes, key components of the core clock oscillator, on body mass, food and water intake, glucose metabolism, and hypothalamic-pituitary-adrenal axis, was investigated in the Per1 ^Brd and Per2 ^Brd mouse models. The results reveal that the lack of Per1 but not Per2 has severe consequences for the regulation of these parameters. Specifically, in Per1 ^Brd animals, we found an impaired daily glucocorticoid rhythm, with markedly elevated levels during the day compared to control animals. In addition, Per1 ^Brd mice showed significant differences in body mass as well as food and water intake. Although the Per1 ^Brd are lighter than wildtype mice, food and water intake per gram body mass is elevated. In addition, the Per1 ^Brd mice exhibit an increased glucose metabolism after i.p. injection with glucose. In conclusion, our study presents first evidence for a link between an altered metabolism in Per1 and Per2 deficient mice, which in the case of the Per1 ^Brd animals might be due to an impaired corticosterone rhythm.     

A strain difference in the daily rhythm of glyceraldehyde-3-phosphate dehydrogenase activity in the mouse

Summary A/J mice differ from C57BL/6J mice in the time of the daily peak of activity of glyceraldehyde-3-phosphate dehydrogenase (GAPD) in thymus and in thyroid. Diurnal rhythms in several other enzymes, and indeed of GAPD in tissues other than thymus and thyroid, were identical in the two strains. The A/J characteristic is dominant, and the trait shows neither X-linked nor Y-linked inheritance.     

A strain difference in the daily rhythm of glyceraldehyde-3-phosphate dehydrogenase activity in the mouse

Journal of Comparative Physiology A - A/J mice differ from C57BL/6J mice in the time of the daily peak of activity of glyceraldehyde-3-phosphate dehydrogenase (GAPD) in thymus and in thyroid....     

Age-related changes in the daily rhythm of photoreceptor functioning and circuitry in a melatonin-proficient mouse strain

Retinal melatonin is involved in the modulation of many important retinal functions. Our previous studies have shown that the viability of photoreceptors and ganglion cells is reduced during aging in mice that lack melatonin receptor type 1. This demonstrates that melatonin signaling is important for the survival of retinal neurons. In the present study, we investigate the effects of aging on photoreceptor physiology and retinal organization in CH3-f+/+ mice, a melatonin proficient mouse strain. Our data indicate that the amplitude of the a and b waves of the scotopic and photopic electroretinogram decreases with age. Moreover, the daily rhythm in the amplitude of the a- and b-waves is lost during the aging process. Similarly, the scotopic threshold response is significantly affected by aging, but only when it is measured during the night. Interestingly, the changes observed in the ERGs are not paralleled by relevant changes in retinal morphological features, and administration of exogenous melatonin does not affect the ERGs in C3H-f(+/+) at 12 months of age. This suggests that the responsiveness of the photoreceptors to exogenous melatonin is reduced during aging.     

The daily rhythm of mice

The house mouse Mus musculus represents a valuable tool for the analysis and the understanding of the mammalian circadian oscillator. Forward and reverse genetics allowed the identification of clock components and the verification of their function within the circadian clockwork. In many cases unforeseen links were discovered between a particular circadian regulatory protein and various diseases or syndromes. Thus, this model system is not only perfectly suited to pinpoint the components of the mammalian circadian clock, but also to unravel metabolic, physiological, and pathological processes linked to the circadian timing system.     

Cholesterol homeostasis in ABCA1/LCAT double-deficient mouse

We examined cholesterol homeostasis in mice with the two major cholesterol transport pathways for catabolism interrupted by disrupting abca1, lcat, or both. Plasma HDL markedly decreased in these genotype but LDL/VLDL decreased only in the double deficiency. Fractional catabolic rate of HDL increased in the order of wild type < abca1(?/?) = lcat(?/?) < abca1(?/?)lcat(?/?). Cholesterol accumulated in the liver by disrupting either gene and more by the double disruption. HDL biogenesis by primary-cultured hepatocytes was negligible in the abca1 deficiency and substantially reduced in the lcat deficiency. Secretion of LDL/VLDL was also decreased in these cells but to a less extent. Cholesterol content in the hepatocytes was in a reciprocal order to lipoprotein generation. Expression of hepatic mRNA of the sterol-related genes reflected the cellular cholesterol increase, such as decrease in SREBP2 and HMG-CoA reductase and increase in apoA-I, apoE, and ABCG1. Cholesterol decreased in the steroidogenic organs by disruption of either gene resulting from low-plasma HDL. Cholesterol in other peripheral tissues generally decreased under normal chow feeding, and interestingly, it was recovered by high-cholesterol feeding, including the cholesterol content in the brain. No apparent vascular lipid deposition was observed in any genotype. Deletion of the two major factors in “reverse cholesterol transport” may not directly result in severe cholesterol transport stagnation in the body of mouse. Other compensatory pathways may back up cholesterol transport among the organs and tissues even when these pathways are impaired.     

Secretory rhythm of growth hormone regulates sexual differentiation of mouse liver

The secretory pattern of growth hormone (GH) differs between the sexes; in males it is more pulsatile than in females. Experiments were performed to test the hypothesis that differences in the secretory rhythm of GH are responsible for sex-dependent liver functions of mice. Continuous delivery of GH was achieved either by introducing metallothionein-GH fusion genes into the germ line or by implanting minipumps. Pulsatile delivery of GH was mimicked by injection. The effects of these treatments on production of hepatic prolactin/GH receptors, albumin, and major urinary protein (MUP) were monitored. The results suggest that induction of MUP mRNA requires pulsatile occupancy of GH receptors, which is achieved naturally in males or by injection of GH, whereas chronic occupancy of GH receptors is inhibitory. In contrast, induction of prolactin/GH receptors requires chronic stimulation of GH receptors, which is approximated in normal female mice or results from increased GH levels in mice with foreign genes or undergoing infusions from minipumps.     

Metabolism of mRNA in regenerating mouse liver: acceleration of the processing and decay of mRNA]

The kinetics of accumulation of poly(A+)mRNA in polyribosomes and the ratio: poly(A+)mRNA/(poly A-)mRNA were studied in regenerating mouse liver. It has been found, that the ratio: (poly A+)mRNA/(poly A-)mRNA was associated with the function of the cells: (poly A+)mRNA fraction has been decreased to 7% at 7 hours after partial hepatectomy and then reached the original value (25%) at 30-40 hours. The kinetics of accumulation of (poly A+)mRNA in polyribosomes during the transition from resting to growing state has revealed that both the lifetime and the presumable time of processing of the mRNAs of free and membranebound polyribosomes were decreased as compared to resting liver cells.     

Marginal iron deficiency enhances liver triglyceride accumulation in rats fed a high-sucrose diet

ABSTRACT

We investigated whether marginal iron-deficiency (MID) without anemia influences liver lipid accumulation in rats. Ingestion of a MID diet in which the iron concentration was half of AIN-93 formulation (iron-adequate, IA) for 3 weeks decreased liver iron concentration without anemia. We then evaluated the influence of the MID diet on liver lipid accumulation in combination with a high-sucrose (HS) diet and confirmed that the HS-MID diet successfully decreased liver iron concentration without anemia. Additionally, a significant increase in liver triglyceride concentration was found, accompanied by upregulation of hepatic fatty acid synthase expression in the rats fed the HS-MID diet compared to those in the rats fed an HS-IA diet, although no difference was observed in plasma transaminase activity and hepatic interleukin-1β expression. These results suggest that MID enhances de novo lipid synthesis via upregulation of lipogenic gene expression in combination with sucrose in the diet.

Abbreviations: ALT, alanine aminotransferase; AST, aspartate aminotransferase; HS, high sucrose; IA, iron adequate; ID, iron deficiency; MID, marginal irondeficiency; NAFLD, non-alcoholic fatty liver disease     

Hepcidin mRNA levels in mouse liver respond to inhibition of erythropoiesis

Hepcidin, a key regulator of iron metabolism, decreases intestinal absorption of iron and its release from macrophages. Iron, anemia, hypoxia, and inflammation were reported to influence hepcidin expression. To investigate regulation of the expression of hepcidin and other iron-related genes, we manipulated erythropoietic activity in mice. Erythropoiesis was inhibited by irradiation or posttransfusion polycythemia and stimulated by phenylhydrazine administration and erythropoietin. Gene expression of hepcidin and other iron-related genes (hemojuvelin, DMT1, ferroportin, transferrin receptors, ferritin) in the liver was measured by the real-time polymerase chain reaction. Hepcidin expression increased despite severe anemia when hematopoiesis was inhibited by irradiation. Suppression of erythropoiesis by posttransfusion polycythemia or irradiation also increased hepcidin mRNA levels. Compensated hemolysis induced by repeated phenylhydrazine administration did not change hepcidin expression. The decrease caused by exogenous erythropoeitin was blocked by postirradiation bone marrow suppression. The hemolysis and anemia decrease hepcidin expression only when erythropoiesis is functional; on the other hand, if erythropoiesis is blocked, even severe anemia does not lead to a decrease of hepcidin expression, which is indeed increased. We propose that hepcidin is exclusively sensitive to iron utilization for erythropoiesis and hepatocyte iron balance, and these changes are not sensed by other genes involved in the control of iron metabolism in the liver.     

Entrainment of circadian locomotor activity rhythm of the nocturnal field mouse Mus booduga using daily injections of melatonin

In this paper, we report the effects of daily injections of melatonin on the locomotor activity rhythm of the nocturnal field mouse Mus booduga. The locomotor activity rhythm of 45 animals was first monitored in constant darkness (DD) of the laboratory for about 15 days. The animals were then divided into three groups (experimental, vehicle-treated control, and the nontreated control groups) and subjected to three different treatments. The animals from the experimental group (n=19) were administered daily a single subcutaneous (s.c.) injection of melatonin (1 mg/kg) for about 45 days. The vehicle treated controls (n=13) were administered daily injections of 50% dimethyl sulfoxide (DMSO) for about 45 days, and the nontreated controls (n=13) were handled similar to the other two groups without being administered injections. Following the treatments, the animals were maintained in DD for about 20 days, after which the experiments were terminated. A significantly larger percentage of animals from the experimental group either entrained or showed phase control to daily treatments, compared to the animals from the two control groups. These results suggest that externally administered melatonin can influence the phase of the circadian locomotor activity rhythm of M. booduga. The fact that none of the nontreated controls showed any sign of phase control to daily handling, clearly demonstrates that the entrainment or phase control in the melatonin treated group of animals is caused by melatonin alone and not due to handling.