A novel mutation in Slc2a4 as a mouse model of fatigue.

Chronic fatigue is a debilitating disorder with widespread consequences, but effective treatment strategies are lacking. Novel genetic mouse models of fatigue may prove invaluable for studying its underlying physiological mechanisms and for testing treatments and interventions. In a screen of voluntary wheel-running behavior in N-ethyl-N-nitrosourea mutagenized C57BL/6J mice, we discovered two lines with low body weights and aberrant wheel-running patterns suggestive of a fatigue phenotype. Affected progeny from these lines had lower daily activity levels and exhibited low amplitude circadian rhythm alterations. Their aberrant behavior was characterized by frequent interruptions and periods of inactivity throughout the dark phase of the light-dark cycle and increased levels of activity during the rest or light phase. Expression of the behavioral phenotypes in offspring of strategic crosses was consistent with a recessive inheritance pattern. Mapping of phenotypic abnormalities showed linkage with a single locus on chromosome 1, and whole exome sequencing identified a single point mutation in the Slc2a4 gene encoding the GLUT4 insulin-responsive glucose transporter. The single nucleotide change (A-T, which we named "twiggy") was in the distal end of exon 10 and resulted in a premature stop (Y440*). Additional metabolic phenotyping confirmed that these mice recapitulate phenotypes found in GLUT4 knockout mice. However, to the best of our knowledge, this is the first time a mutation in this gene has been shown to result in extensive changes in general behavioral patterns. These findings suggest that GLUT4 may be involved in circadian behavioral abnormalities and could provide insights into fatigue in humans.

Mice under Caloric Restriction Self-Impose a Temporal Restriction of Food Intake as Revealed by an Automated Feeder System.

Caloric restriction (CR) extends lifespan in mammals, yet the mechanisms underlying its beneficial effects remain unknown. The manner in which CR has been implemented in longevity experiments is variable, with both timing and frequency of meals constrained by work schedules. It is commonplace to find that nocturnal rodents are fed during the daytime and meals are spaced out, introducing prolonged fasting intervals. Since implementation of feeding paradigms over the lifetime is logistically difficult, automation is critical, but existing systems are expensive and not amenable to scale. We have developed a system that controls duration, amount, and timing of food availability and records feeding and voluntary wheel-running activity in mice. Using this system, mice were exposed to temporal or caloric restriction protocols. Mice under CR self-imposed a temporal component by consolidating food intake and unexpectedly increasing wheel-running activity during the rest phase, revealing previously unrecognized relationships among feeding, metabolism, and behavior.

FGF21 regulates metabolism and circadian behavior by acting on the nervous system.

Fibroblast growth factor 21 (FGF21) is a hepatokine that acts as a global starvation signal to modulate fuel partitioning and metabolism and repress growth; however, the site of action of these diverse effects remains unclear. FGF21 signals through a heteromeric cell-surface receptor composed of one of three FGF receptors (FGFR1c, FGFR2c or FGFR3c) in complex with ß-Klotho, a single-pass transmembrane protein that is enriched in metabolic tissues. Here we show that in addition to its known effects on peripheral metabolism, FGF21 increases systemic glucocorticoid levels, suppresses physical activity and alters circadian behavior, which are all features of the adaptive starvation response. These effects are mediated through ß-Klotho expression in the suprachiasmatic nucleus of the hypothalamus and the dorsal vagal complex of the hindbrain. Mice lacking the gene encoding ß-Klotho (Klb) in these regions are refractory to these effects, as well as those on metabolism, insulin and growth. These findings demonstrate a crucial role for the nervous system in mediating the diverse physiologic and pharmacologic actions of FGF21.

Housing conditions influence the expression of food-anticipatory activity in mice.

If food is presented for a limited duration once every 24 h, rats will gradually develop increased activity in the hours preceding mealtime, even if this is during a time of day when they are normally inactive. This food-anticipatory activity (FAA) is mediated by the entrainment of an endogenous self-sustaining circadian pacemaker. Mice are increasingly being used for the molecular analysis of behavior because of the large number of genetically manipulated mouse models available, but little is known about food entrainment in this species. This study assessed the impact of different housing conditions on the expression of FAA in a temporally restricted feeding paradigm. Wheel-running activity rhythms were recorded from mice (Mus musculus) undergoing food restriction while housed under one of two conditions. The results demonstrated that mice housed on open shelves showed robust FAA while those housed in isolation boxes did not. These results indicate that differences in susceptibility to food entrainment among mice (e.g., those with different genotypes) should be interpreted cautiously, since at least one strain is strongly affected by a relatively minor procedural difference that was not anticipated to have a significant impact.

Entrainment impaired, masking spared: an apparent genetic abnormality that prevents circadian rhythm entrainment to 24-h lighting cycles in California mice.

Lighting cycles can influence the expression of daily activity rhythms in two ways: by entraining the circadian pacemaker that normally drives this rhythm, and by directly affecting the expression of activity itself, thereby 'masking' the influence of the pacemaker. We describe a California mouse (Peromyscus californicus) in which these processes are dissociated. Circadian rhythms of wheel-running activity were recorded continuously while the animal was housed in a standard light/dark cycle and in constant darkness. This animal expressed a normal circadian rhythm that failed to entrain to the light/dark cycle, but was completely masked during the light phase. This animal's phenotype appears to have a genetic basis, since the progeny of selective crosses of his descendants showed similar abnormalities. These mice are the first example of animals expressing apparently normal circadian rhythms that are not entrained by light, but that still show potent masking responses to light exposure.