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The purinergic type P1 (adenosine A 1  and A 2A ) receptors and the type P2 (X7) receptor have been suggested to mediate physiological effects of adenosine and adenosine triphosphate on sleep. We aimed to determine gene expression of A 1 R (receptor), A 2A R, and P2RX 7  in leukocytes of healthy subjects during total sleep deprivation followed by sleep recovery. Expression of the pro-inflammatory cytokines IL-1β and TNF-α were also determined as they have been characterized as sleep regulatory substances, via P2RX 7  activation. Blood sampling was performed on 14 young men (aged 31.9 ± 3.9) at baseline (B), after 24 h of sleep deprivation (24 h- SD ), and after one night of sleep recovery (R). We compared gene expression levels after six nights of habitual (22.30–07.00) or extended (21.00–07.00) bedtimes. Using quantitative real-time PCR, the amount of mRNA for A 1 R, A 2A R, P2RX 7 , TNF-α, and IL-1β was analyzed. After 24 h- SD  compared to B, whatever prior sleep condition, a significant increase of A 2A R expression was observed that returned to basal level after sleep recovery [day main effect,  F  (2, 26)  = 10.8,  p  < 0.001]. In both sleep condition, a day main effect on P2RX 7  mRNA was observed [ F  (2, 26)  = 6.7,  p  = 0.005] with significant increases after R compared with 24 h- SD . TNF-α and IL-1β expressions were not significantly altered. Before 24 h- SD  (baseline), the A 2A R expression was negatively correlated with the latency of stage 3 sleep during the previous night, while that of the A 1 R positively. This was not observed after sleep recovery following 24 h- SD . This is the first study showing increased A 2A R and not A 1  gene expression after 24 h- SD  in leukocytes of healthy subjects, and this even if bedtime was initially increased by 1.5 h per night for six nights. In conclusion, prolonged wakefulness induced an up-regulation of the A2A receptor gene expression in leukocytes from healthy subjects. Significant correlations between baseline expression of A 1  and A 2A  receptors in peripheral cells and stage 3 sleep suggested their involvement in mediating the effects of adenosine on sleep.

frontiers in neuroscience

Leukocyte Expression of Type 1 and Type 2 Purinergic Receptors and Pro-Inflammatory Cytokines during Total Sleep Deprivation and/or Sleep Extension in Healthy Subjects