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. 2021 Apr 9;12:2113. doi: 10.1038/s41467-021-22179-z

Fig. 5. Altered levels of endogenous adenosine regulate circadian rhythms in vivo.

Fig. 5

a Basal forebrain (BF) and SCN were collected from wild type and human adenosine kinase overexpressing transgenic mice (Adk-Tg - ADK) at indicated circadian times. Each sample is a pool of 5–7 individual tissue punches. b Phase shifting in Adk-Tg animals (ADK-Tg) and wild type (WT) in response to a 1 h 200 lux light pulse at CT14 (n = 10 and 9, no significant difference, t-test) (b) and CT22 (c) (n = 14 and 8, p = 0.0439, t-test). d Free running period of the same mice in DD (n = 12 and 9, p = 0.0071). * = p < 0.05, ** = p < 0.01 t-test. e Behavioural phase-shifts in response to a 30 min 20 lux light pulse (LP) at CT14 (n = 14) is attenuated by sleep deprivation (SD, n = 10) for the previous 6 h (p = 0.0001, n = 9–11), but potentiated when pretreated with 5 mg/kg JNJ (n = 9, p = 0.0007) one-way ANOVA with Tukey multiple comparisons. f Sleep deprivation (ZT6-12) reduces the expression of Per1 (n = 11,11, t-test, p = 0.031) and g Per2 mRNA (n = 12,12 t-test, p = 0.025) within the SCN. h JNJ (5 mg/kg i.p. administered at ZT6) reverses the attenuation of Per2 within the SCN (n = 9,11, 8, p = 0.0006, one-way ANOVA with Dunnett’s multiple test correction). i–m Behavioural phase shifts in response to light and drugs that modulate adenosine signalling via specific A1 or A2A receptor agonists or antagonists: i N6CPA (A1 agonist, 0.5 mg/kg, p = 0.0164) and CGS21680 (A2A agonist, 0.5 mg/kg, p = 0.0046) (n = 12, 9, 13) and j KW3902 (A1 antagonist, 1 mg/kg, p = 0.0051) and KW6002 (A2A antagonist, 1 mg/kg) (n = 11, 9, 11). In i and j the phase-delaying phase shifts induced by a light pulse (30 min, 10 lux) delivered at CT16 are shown. For comparison, the phase-advance induced by a 30 min 100 lux light pulse at CT22 is shown in k (n = 10, 9, 10) and l (n = 10, 8, 8) (p = 0.0188 for KW3902 and p = 0.0078 for KW6002. One-way ANOVA for i–l, Dunnett’s multiple test correction. Tukey’s box plots used throughout (central line – mean box represents 25th to 75th percentile data, whiskers are 1.5 interquartile range). m Representative actograms are shown for i–I.