The 5-week old tim01 mutant flies subjected to ad libitum feeding (ARF) have a slower heart rate compared to controls. The reduction in arrhythmia on 12-hour daytime-restricted feeding (TRF) seen in wild type is not observed in tim01 mutant flies.
tim01 mutants exhibit normal light-dependent temperature preference. Similarly to wild-type, they prefer higher temperature in the light than in the dark.
Total sleep at nighttime and daytime is significantly decreased in tim01 mutants compared with wild-type. This is caused mainly by sleep episode duration.
Sleep bout duration and sleep latency are significantly reduced compared to controls in mutant flies.
tim01 mutant flies exhibit arrhythmic temperature preference rhythm in constant darkness after two days and in constant light at day 4. There is also a 'masking' effect of the light-dark cycle, in which tim01 flies prefer a higher temperature during the day (ZT1-12) and a lower temperature during the night (ZT13-24). tim01 flies kept in constant light for 4 days prefer a higher temperature during the daytime than flies kept in constant darkness for 2 days.
During the daytime, tim01 flies constantly prefer a temperature of approximately 26.5[o]C and do not exhibit a daytime temperature preference rhythm in light-dark conditions.
Triglyceride levels in tim01 mutants are similar to wild-type.
tim01 homozygotes exhibit aberrant locomotor rhythm under conditions of constant darkness.
tim01; timT:Avic\GFP-YFP flies do not show significant changes in period at different temperatures.
Naive and trained responses to a magnetic field are not impaired in tim01 mutant flies.
In contrast to w1118 controls, tim01 larvae show neutral performance in phototaxis tests, prefer light to darkness in the immediate light/dark boundary passing test, and demonstrate no significant motility alteration in the dark condition.
No significant difference is found in the number of dendritic branches between ZT2 and ZT14 in tim01 mutants.
16.7% of timZ2-3326/tim01 flies are arrhythmic under constant darkness conditions. The period of rhythmic mutant flies is 28.7 +/- 0.28 hours, longer than that of controls.
The rhythmic change in bouton size that is seen in motor neuron MN5 under LD (12 hour light:12 hour dark) conditions and DD (constant darkness) conditions in wild-type flies is abolished in mutant flies.
Motor neuron MN5 shows increased neuronal branching in mutant flies compared to wild type. This increased branching is associated with axonal abnormalities such as thickenings, sprouting and coiling in high-order branches.
tim01 females show a significantly higher level of ovarian diapause compared to wild-type females.
tim01 has no effect on experience-dependent responses in sleep need.
tim01 flies driven by a light:dark cycle show a steady decline in sleep:wake rhythm strength with age.
tim01 flies have poorly consolidated sleep compared with wild type flies: mutant flies have more sleep bouts and a shorter sleep bout duration. There is no substantial increase in the sleep fragmentation of tim01 flies with age: although sleep bout numbers increase somewhat with age, the average duration of sleep bouts does not increase.
tim01 mutants exhibit no circadian regulation of locomotor behaviour under light-dark conditions, other than a preference for being active during the light phase.
tim01 mutant larvae are insensitive to light in light avoidance assays, even at high light intensity (750 lux). Increasing the light to 1100 lux partially rescues the photophobic defects of tim01 mutants, indicating that these mutants are not completely blind and that the molecular clock regulates visual sensitivity.
Homozygotes are arrhythmic for locomotor activity under free-running conditions.
Mutant flies are arrhythmic for locomotor activity under constant light conditions at a constant temperature of 25[o]C, but become rhythmic immediately after being exposed to a temperature cycle of 12 hours at 25[o]C and 12 hours at 30[o]C. Peaks in locomotor activity occur at temperature transitions. The locomotor activity rhythm of the mutant flies differs from that of wild-type flies, in that neither anticipatory activity nor transients after a transfer to temperature cycles is observed.
The time spent copulating is significantly increased in tim01 mutant males mated to wild-type females, compared to wild-type males. This effect is seen in both 2 day old and 4 day old males. This increased time in copulation does not appear to be due to difficulty of males disengaging from females. The time spent copulating by tim01/tim03 males mated with wild type females is significantly extended compared to wild-type.
Long term memory of courtship conditioning (reduction in time spent in courtship behaviour 5 days after a 7 hour conditioning) is normal in tim01 homozygous males.
tim01 flies are arrhythmic with respect to locomotor activity under constant darkness conditions.
Single tim01 mutant females mated once to single tim01 mutant males lay significantly less eggs and produce significantly less progeny than wild-type flies. The percentage of unfertilised eggs from this mating is significantly greater than for wild-type. These effects on fertility are significantly reduced if pairs of per01 mutant flies are allowed to mate over a period of 4 days. Single wild-type females mated once to single tim01 mutant males lay significantly less eggs and produce significantly less progeny than those mated to wild-type males. The numbers of sperm released to the seminal vesicles by tim01 males over the 2 days following eclosion is significantly less than that seen in wild-type males. Single tim01/tim03 females mated once to single tim01/tim03 males produce significantly less progeny than do wild-type flies, even though the number of eggs laid and the percentage of fertilised eggs is not significantly different to wild-type. tim01/tim03 males have significantly less sperm in their seminal vesicles 42 hours after eclosion than wild-type males.
Mutant females show altered sleep rebound after sleep deprivation of 7, 9 or 12 hours (in constant darkness) compared to wild-type flies.
tim01/tim03 flies show arrhythmic locomotor activity.
96% of tim01 flies are arrhythmic.
When wild-type flies are moved from a LD cycle to a DD cycle, at CT12 (circadian time) there is a reduction in mating activity seen. This reduction is abolished in tim01 mutants. If either male or females are mutant, this abolition of mating rhythm is seen, even if they are mated to wild-type flies.
The circadian rhythms seen in wild-type cultured malphighian tubules and rectums are essentially eliminated.
The baseline rest patterns of tim01 flies studied in constant darkness conditions are arrhythmic. After rest deprivation at circadian time (CT) 18-24, tim01 flies show a significant decrease in rest, similar to handled control wild-type flies. tim01 flies significantly decrease rest after deprivation compared to handled tim01 controls.
Homozygotes are have completely arrhythmic circadian cycles.
Heterozygotes have a locomotion period indistinguishable from wild-type flies.
Unlike other circadian gene mutants, tim01 flies show a wild-type response to cocaine exposure.
Circadian rhythms in olfactory responses to ethyl acetate or benzaldehyde are abolished in tim01 mutants.
Behaviorally arrhythmic at 18oC, 25oC and 29oC. The molecular program is stuck at about ZT 7-8.
Homozygous flies are arrhythmic with respect to locomotor activity.
Only 9% of flies are arrhythmic with respect to locomotor activity under 12 hour light:12 hour dark conditions.
Exposure of flies to constant light causes a tim01 phenocopy i.e. behavioral arrhythmia.
Individuals are arrhythmic.
Altered eclosion rhythm, night-emerging and day-emerging adults. Defects in locomotor activity, flies become arrhythmic when the entraining light-dark cues are absent. No detectable defect in nervous system, visual system or brain.
Aberrant behavioural phenotype and localisation of per.