Most organisms, including humans,exhibit daily rhythms in their biological activities, physiological functions, and homeostatic mechanisms such as cell regeneration, hormone production, cardiac output, blood pressure, blood flow distribution, and body temperature. The physiological system responsible for these rhythms is known as the circadian system. Circadian changes have increasingly become an interesting focus of research, concerning also neurobehavioral functioning of healthy subjects. The impact of factors such as the sleep–wakefulness cycle and biological time-of-day on measures of subjective alertness has been extensively studied (1, 2). Moreover, there is compelling evidence of circadian dependency also for cognitive functions such as attention, memory, and learning (3, 4). Recently, it has been consolidated in different experimental models, including mammalian brain, that the circadian clock has a role in regulating structural synaptic plasticity, opening the new relevant concern of circadian-dependent neural plasticity (5,6). Interestingly, it has been suggested that changes in the electrical properties of the cell membrane (intrinsic plasticity) and in the release of neuromodulatory molecules due to the internal clock can reconfigure circuit dynamics leading individual neurons to switch among different functional networks throughout the day (5). Daily rhythmicity in neural activity has been further elucidated by Blautzik et al. (7) who analyzed the daily course of connectivity patterns. The authors found different degrees of daily modulation across connectivity patterns, ranging from networks characterized by stable activity across the day and networks with highly rhythmic connectivity changes. Based on the reported findings, we can infer that the aforementioned oscillatory processes in connectivity strength and spatial extent would eventually determine highly individual fluctuations of effective connectivity over the course of the day. Circadian rhythms show also to exert influence on the excitability of the cerebral cortex, as found by Lang et al. (8). In this study, the excitability of the primary motor cortex (M1) of healthy subjects was evaluated by transcranial magnetic stimulation (TMS) at different times of the day. Data unveiled that both the intracortical and the corticospinal excitability of M1 exhibited a progressive decrease during the course of the day. In the last years, it has emerged that the effectiveness and reproducibility of several techniques able to induce neuroplastic changes in humans, such as paired associative stimulation (PAS), are influenced by time-of-day of the intervention (9) and subjected to circadian modulation. As demonstrated by Sale et al. (10), who tested 25 subjects twice, at 8:00 a.m. and 8:00 p.m., on separate days, PAS effectiveness is enhanced in the evening,when endogenous cortisol is low; conversely, effects of PAS in the evening are blocked by a single oral dose of hydrocortisone. Additionally, in a large study of humans aged 50–70 years, high salivary levels of cortisol appeared to be related with poor performances on a wide range of cognitive domains, including language, verbal learning, processing speed, memory, and eye–hand coordination (11). Overall, the circadian modulation of several neural properties and structures, at both the microscopic and functional levels, may deeply affect cognitive behavior, responsiveness, and performance within the day. Despite its potential impact, timeof-day is rarely contemplated when brain responses and cognitive functions are studied. As a matter of fact, in order to minimize possible biases related to circadian effects, some neurophysiological studies are conducted with evaluations and/or interventions performed at the same time of the day. Still, these experimental designs do not contemplate another relevant factor, which may strongly affect the reliability of the collected data that is the interindividual variability of the biological clock. This physiological variability of circadian rhythms between subjects has led to the notion of individual chronotypes (12, 13). The chronotype influences the organization of physiological functions, behaviors, and cognitive performances throughout the day (1, 14, 15). Given the differences in circadian rhythmicity between chronotypes, specific individual variations in task performance are likely to occur as a function of time-of-day. In other words, this implies that the scheduled task may not be necessarily synchronized to the most optimal moment in the day for each tested participant (16, 17). The regulation of the endogenous dynamics that characterizes a chronotype is dictated by many factors. The thorough understanding of these mechanisms is critical to gain a comprehensive view on their functional implications and, further, may be considerably useful when addressing the experimental limitations due to diurnal rhythmicity. Among them, cortisol is a main neuromodulator that mediates circadian processes. The normal diurnal pattern of cortisol secretion has been
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