Movements during sleep reveal the developmental emergence of a cerebellar-dependent internal model in motor thalamus.

With our eyes closed, we can track a limb's moment-to-moment location in space. If this capacity relied solely on sensory feedback from the limb, we would always be a step behind because sensory feedback takes time: for the execution of rapid and precise movements, such lags are not tolerable. Nervous systems solve this problem by computing representations-or internal models-that mimic movements as they are happening, with the associated neural activity occurring after the motor command but before sensory feedback. Research in adults indicates that the cerebellum is necessary to compute internal models. What is not known, however, is when-and under what conditions-this computational capacity develops. Here, taking advantage of the unique kinematic features of the discrete, spontaneous limb twitches that characterize active sleep, we captured the developmental emergence of a cerebellar-dependent internal model. Using rats at postnatal days (P) 12, P16, and P20, we compared neural activity in the ventral posterior (VP) and ventral lateral (VL) thalamic nuclei, both of which receive somatosensory input but only the latter of which receives cerebellar input. At all ages, twitch-related activity in VP lagged behind the movement, consistent with sensory processing; similar activity was observed in VL through P16. At P20, however, VL activity no longer lagged behind movement but instead precisely mimicked the movement itself; this activity depended on cerebellar input. In addition to demonstrating the emergence of internal models of movement, these findings implicate twitches in their development and calibration through, at least, the preweanling period.

Forebrain-independent generation of hyperthermic convulsions in infant rats.

Febrile seizures are the most common type of convulsive events in children. It is generally assumed that the generalization of these seizures is a result of brainstem invasion by the initial limbic seizure activity. Using precollicular transection in 13-day-old rats to isolate the forebrain from the brainstem, we demonstrate that the forebrain is not required for generation of tonic-clonic convulsions induced by hyperthermia or kainate. Compared with sham-operated littermate controls, latency to onset of convulsions in both models was significantly shorter in pups that had undergone precollicular transection, indicating suppression of the brainstem seizure network by the forebrain in the intact animal. We have shown previously that febrile seizures are precipitated by hyperthermia-induced respiratory alkalosis. Here, we show that triggering of hyperthermia-induced hyperventilation and consequent convulsions in transected animals are blocked by diazepam. The present data suggest that the role of endogenous brainstem activity in triggering tonic-clonic seizures should be re-evaluated in standard experimental models of limbic seizures. Our work sheds new light on the mechanisms that generate febrile seizures in children and, therefore, on how they might be treated.

Development of SCN connectivity and the circadian control of arousal: a diminishing role for humoral factors?

The suprachiasmatic nucleus (SCN) is part of a wake-promoting circuit comprising the dorsomedial hypothalamus (DMH) and locus coeruleus (LC). Although widely considered a "master clock," the SCN of adult rats is also sensitive to feedback regarding an animal's behavioral state. Interestingly, in rats at postnatal day (P)2, repeated arousing stimulation does not increase neural activation in the SCN, despite doing so in the LC and DMH. Here we show that, by P8, the SCN is activated by arousing stimulation and that selective destruction of LC terminals with DSP-4 blocks this activational effect. We next show that bidirectional projections among the SCN, DMH, and LC are nearly absent at P2 but present at P8. Despite the relative lack of SCN connectivity with downstream structures at P2, day-night differences in sleep-wake activity are observed, suggesting that the SCN modulates behavior at this age via humoral factors. To test this hypothesis, we lesioned the SCN at P1 and recorded sleep-wake behavior at P2: Day-night differences in sleep and wake were eliminated. We next performed precollicular transections at P2 and P8 that isolate the SCN and DMH from the brainstem and found that day-night differences in sleep-wake behavior were retained at P2 but eliminated at P8. Finally, the SCN or DMH was lesioned at P8: When recorded at P21, rats with either lesion exhibited similarly fragmented wake bouts and no evidence of circadian modulation of wakefulness. These results suggest an age-related decline in the SCN's humoral influence on sleep-wake behavior that coincides with the emergence of bidirectional connectivity among the SCN, DMH, and LC.

Developmental appearance and disappearance of cortical events and oscillations in infant rats.

Until recently, organized and state-dependent neocortical activity in infant rats was thought to commence with the emergence of delta waves at postnatal day (P)11. This view is changing with the discovery of several forms of cortical activity that are detectable soon after birth, including spindle bursts (SBs) and slow activity transients (SATs). Here we provide further evidence of surprisingly rich cortical activity patterns during early development and document, in P5-P13 rats, the appearance, disappearance, and transient expression of three cortical events and oscillations. EEG activity in frontal, parietal, and occipital cortices was recorded in unanesthetized, head-fixed subjects using 16-channel laminar silicon electrodes and Ag-AgCl electrodes. In addition to SATs, we identified two novel forms of activity: cortical sharp potentials (CSPs) and gamma bursts (GBs). SBs were not observed in these areas. CSPs, defined as discrete, biphasic events with a duration of 250 ms, exhibited an inverted-U developmental trajectory with peak prevalence at P9. In contrast, GBs, defined as brief bursts of 40-Hz activity, increased steadily in prevalence and duration from P5 through P13. The prevalence of SATs decreased steadily across the ages tested here. Furthermore, both CSPs and GBs were more likely to occur during sleep than during wakefulness. Because SATs, CSPs, and GBs exhibit different developmental trajectories and rates of occurrence, and can occur independently of each other, they appear to be distinct patterns of neuronal activity. We hypothesize that these diverse patterns of neurophysiological activity reflect the instantaneous local structure and connectivity of the developing neocortex.

Brainstem and hypothalamic regulation of sleep pressure and rebound in newborn rats.

Sleep pressure and rebound comprise the two compensatory or "homeostatic" responses to sleep deprivation. Although sleep pressure is expressed by infant rats as early as postnatal day (P)5, sleep rebound does not appear to emerge until after P11. We reexamined the developmental expression of these sleep-regulatory processes in P2 and P8 rats by depriving them of sleep for 30 min using a cold, arousing stimulus delivered to a cold-sensitive region of the snout. This method effectively increased sleep pressure over the 30-min period (i.e., increases in the number of arousing stimuli presented over time). Moreover, sleep rebound (i.e., increased sleep during the recovery period) is demonstrated for the first time at these ages. Next, we showed that precollicular transections in P2 rats prevent sleep rebound without affecting sleep pressure, suggesting that the brainstem is sufficient to support sleep pressure, but sleep rebound depends on neural mechanisms that lie rostral to the transection. Finally, again in P2 rats, we used c-fos immunohistochemistry to examine neural activation throughout the neuraxis during sleep deprivation and recovery. Sleep deprivation and rebound were accompanied by significant increases in neural activation in both brainstem and hypothalamic nuclei, including the ventrolateral preoptic area and median preoptic nucleus. This early developmental expression of sleep pressure and rebound and the apparent involvement of brainstem and hypothalamic structures in their expression further solidify the notion that sleep-wake processes in newborns-defined at these ages without reference to state-dependent EEG activity-provide the foundation on which the more familiar processes of adults are built.