Behavioral correlates of direct current-coupled electrographic activity in premature infants.

The co-expression of behavioral and neural events represents a situation conducive to Hebbian-type neuroplasticity and may provide a reasonable explanation for how the amount of movement during the perinatal period contributes to neuromotor development. Direct current-coupled electrographic recordings in premature infants indicate that the majority of the electrographic activity is exhibited in a slow frequency range that is either distorted or not visible using traditional recording methods. Therefore, we provide a description of the behavioral correlates of direct current-coupled electrographic recordings in six premature human infants (3 males and 3 females; 30-34 weeks). We report, in concert with prior data, that electrographic activity and movements occur in tightly coupled discrete bouts. Surprisingly, spontaneous activity transients, which are slow, high amplitude, multiband electrographic events, typically precede startles; thereby revealing a previously unknown coupling of early neural and behavioral events in humans. Taken together, the present findings open novel venues for studying and dissecting mammalian neuromotor development.

Developmental emergence of transient and persistent hippocampal events and oscillations and their association with infant seizure susceptibility.

During the second postnatal week in rats, the hippocampus exhibits a transient period of hyperexcitability. To systematically assess the relationship between the onset and end of this period and spontaneous hippocampal activity, we used silicon depth electrodes in unanaesthetized head-fixed rats from postnatal day (P)2 to P18. At all ages, hippocampal sharp waves (SPWs) were prominent in the EEG. Beginning at P6, however, marked changes in SPWs and associated oscillations were detected. SPW-related 'gamma tails' (60-100 Hz) and 'ripples' (140-200 Hz) were first observed at P6 and P7, respectively, and both oscillations persisted up to P18. Transiently, between P6 and P11, SPW duration decreased and the occurrence of SPW doublets increased. In addition, between P8 and P11, a subset of rats exhibited 'spontaneous potentiated SPWs' characterized by double polarity reversals, enhanced likelihood of gamma tails, and population spikes. Having identified a suite of transient hippocampal features consistent with a window of increased excitability, we next assessed whether electrographic seizure activity would be most easily induced during this period. To do this, kainic acid (KA; 200 ng/infusion) was infused into the hippocampus contralateral to the recording probe. KA did not induce seizure activity until P7 and reached peak effectiveness at P9. Thereafter, sensitivity to KA declined. All together, these findings provide in vivo neurophysiological support for the notion of a developmental window of heightened seizure susceptibility during the second postnatal week, and also suggest that spontaneous nonpathological hippocampal activity can be used to mark the onset and end of this period.

On the co-occurrence of startles and hippocampal sharp waves in newborn rats.

Hippocampal sharp waves (SPWs) are among the earliest neural population patterns observed in infant mammals. Similarly, startles are among the earliest behavioral events observed. Here we provide evidence indicating that these two events are linked mechanistically soon after birth in freely moving and head-fixed 1 to 4-day-old rats. EMG electrodes and intrahippocampal silicon depth electrodes were used to detect the presence of startles and SPWs, respectively. In intact pups, the majority of sharp waves were preceded by startles (average latency: 161 ms). When the hippocampal formation was surgically separated from the brainstem, however, sharp waves and startles still occurred, but now independently. In addition, unrelated to startles or SPWs, gamma oscillations were detected in several subjects, as were neocortical "spindles" that propagated passively into the hippocampus. The co-occurrence of sharp waves and startles provides the opportunity for Hebbian changes in synaptic efficacy and, thus, is poised to contribute to the assembly of neural circuits early in development.

The preoptic hypothalamus and basal forebrain play opposing roles in the descending modulation of sleep and wakefulness in infant rats.

Recent findings in infant rats suggest that the preoptic area (POA) and/or basal forebrain (BF) contribute to developmental changes in sleep and wake organization between postnatal day 2 (P2) and P9. To examine the contributions of these forebrain areas to sleep and wakefulness, separate lesions of the POA or BF, or combined lesions (POA + BF), were performed at P9, and precollicular transections were performed at P2. In addition, modafinil, a drug of unknown mechanism of action the effects of which on sleep and wakefulness have been hypothesized to result from inhibition of POA activity, was administered at P2 and P9. Finally, extracellular neuronal activity was recorded from the POA and BF. POA lesions decreased sleep bout durations and increased wake bout durations. BF lesions inhibited sleep bout durations to a lesser extent, while leaving wake bout durations unaffected. POA + BF lesions produced a combination of these effects, resulting in short bouts of sleep and wakefulness similar to those of transected P8 rats. Even at P2, transections decreased sleep bout durations. The finding, however, that the sleep-inhibiting and wake-promoting effects of modafinil were more potent at P9 than at P2 suggests increasing sleep-wake modulation by the POA between these two ages. Finally, neuronal recordings confirmed the presence of state-dependent neurons within the infant POA and BF. We propose that the POA, in addition to promoting sleep, inhibits wakefulness via direct and indirect inhibitory connections with wake-promoting neurons in the BF, and that this inhibitory influence increases across early development.

Dynamics of sleep-wake cyclicity in developing rats.

Adult mammals cycle between periods of sleep and wakefulness. Recent assessments of these cycles in humans and other mammals indicate that sleep bout durations exhibit an exponential distribution, whereas wake bout durations exhibit a power-law distribution. Moreover, it was found that wake bout distributions, but not sleep bout distributions, exhibit scale invariance across mammals of different body sizes. Here we test the generalizability of these findings by examining the distributions of sleep and wake bout durations in infant rats between 2 and 21 days of age. In agreement with Lo et al., we find that sleep bout durations exhibit exponential distributions at all ages examined. In contrast, however, wake bout durations also exhibit exponential distributions at the younger ages, with a clear power-law distribution only emerging at the older ages. Further analyses failed to find substantial evidence either of short- or long-term correlations in the data, thus suggesting that the durations of current sleep and wake bouts evolve through time without memory of the durations of preceding bouts. These findings further support the notion that bouts of sleep and wakefulness are regulated independently. Moreover, in light of recent evidence that developmental changes in sleep and wake bouts can be attributed in part to increasing forebrain influences, these findings suggest the possibility of identifying specific neural circuits that modulate the changing complexity of sleep and wake dynamics during development.

Extraocular muscle activity, rapid eye movements and the development of active and quiet sleep.

Rapid eye movements (REMs), traditionally measured using the electrooculogram (EOG), help to characterize active sleep in adults. In early infancy, however, they are not clearly expressed. Here we measured extraocular muscle activity in infant rats at 3 days of age (P3), P8 and P14-15 in order to assess the ontogeny of REMs and their relationship with other forms of sleep-related phasic activity. We found that the causal relationship between extraocular muscle twitches and REMs strengthened during the first two postnatal weeks, reflecting increased control of the extraocular muscles over eye movements. As early as P3, however, phasic bursts of extraocular muscle twitching occurred in synchrony with twitching in other muscle groups, producing waves of phasic activity interspersed with brief periods of quiescence. Surprisingly, the tone of the extraocular muscles, invisible to standard EOG measures, fluctuated in synchrony with the tone of other muscle groups; focal electrical stimulation within the dorsolateral pontine tegmentum, an area that has been shown to contain wake-on neurons in P8 rats, resulted in the simultaneous activation of high tone in both nuchal and extraocular muscles. Finally, when state-dependent neocortical electroencephalographic activity was observed at P14, it had already integrated fully with sleep and wakefulness as defined using electromyographic criteria alone; this finding is not consistent with the notion that active sleep in infants at this age is 'half-activated.' All together, these results indicate exquisite temporal organization of sleep soon after birth and highlight the possible functional implications of homologous activational states in striated muscle and neocortex.

The neural substrates of infant sleep in rats.

Sleep is a poorly understood behavior that predominates during infancy but is studied almost exclusively in adults. One perceived impediment to investigations of sleep early in ontogeny is the absence of state-dependent neocortical activity. Nonetheless, in infant rats, sleep is reliably characterized by the presence of tonic (i.e., muscle atonia) and phasic (i.e., myoclonic twitching) components; the neural circuitry underlying these components, however, is unknown. Recently, we described a medullary inhibitory area (MIA) in week-old rats that is necessary but not sufficient for the normal expression of atonia. Here we report that the infant MIA receives projections from areas containing neurons that exhibit state-dependent activity. Specifically, neurons within these areas, including the subcoeruleus (SubLC), pontis oralis (PO), and dorsolateral pontine tegmentum (DLPT), exhibit discharge profiles that suggest causal roles in the modulation of muscle tone and the production of myoclonic twitches. Indeed, lesions in the SubLC and PO decreased the expression of muscle atonia without affecting twitching (resulting in "REM sleep without atonia"), whereas lesions of the DLPT increased the expression of atonia while decreasing the amount of twitching. Thus, the neural substrates of infant sleep are strikingly similar to those of adults, a surprising finding in light of theories that discount the contribution of supraspinal neural elements to sleep before the onset of state-dependent neocortical activity.

The ontogeny of mammalian sleep: a response to Frank and Heller (2003).

In a recent review, Frank and Heller (2003) provided support for their 'presleep theory' of sleep development. According to this theory, rapid eye movement (REM) and non-rapid eye movement (Non-REM) sleep in rats emerge from a common 'dissociated' state only when the neocortical EEG differentiates at 12 days of age (P12). Among the assumptions and inferences associated with this theory is that sleep before EEG differentiation is only 'sleep-like' and can only be characterized using behavioral measures; that the neural mechanisms governing presleep are distinct from those governing REM and Non-REM sleep; and that the presleep theory is the only theory that can account for developmental periods when REM and Non-REM sleep components appear to overlap. Evidence from our laboratory and others, however, refutes or casts doubt on these and other assertions. For example, infant sleep in rats is not 'sleep-like' in that it satisfies nearly every criterion used to characterize sleep across species. In addition, beginning as early as P2 in rats, myoclonic twitching occurs only against a background of muscle atonia, indicating that infant sleep is not dissociated and that electrographic measures are available for sleep characterization. Finally, improved techniques are leading to new insights concerning the neural substrates of sleep during early infancy. Thus, while many important developmental questions remain, the presleep theory, at least in its present form, does not accurately reflect the phenomenology of infant sleep.

Temperature-induced reciprocal activation of hippocampal field activity.

Hippocampal network activity oscillates between sustained rhythms (e.g., theta) and aperiodic population spikes (e.g., sharp waves, dentate spikes). Although temperature is known to modulate various aspects of rhythmic hippocampal activity, little is known regarding the influence of temperature on the incidence of population spikes. We recorded spontaneous hippocampal activity along the CA1-dentate gyrus axis using multisite silicon electrodes in urethanized infant rats (P2-P16) at brain temperatures of 37 and 27 degrees C. Theta and gamma activity, as well as sharp waves, were detected at 37 degrees C but not at 27 degrees C. In contrast, dentate spikes were rare at 37 degrees C but their incidence increased several-fold at 27 degrees C (epileptiform activity also emerged at 27 degrees C in the oldest pups). This surprising increase in the incidence of dentate spike activity in a cold brain represents the first such demonstration for a neuronal field pattern. In addition, these findings indicate that changes in brain temperature produce systems-level shifts in the balance among reciprocally interacting hippocampal components.

The union of the state: myoclonic twitching is coupled with nuchal muscle atonia in infant rats.

Active sleep (AS), as measured by the occurrence of myoclonic twitching (MT), is the most prevalent behavioral state in newborn rats. Historically, AS has been considered a developmental precursor of REM sleep, but recently this idea has been questioned. In the present study, the authors assess, in 2-, 5-, and 8-day-old rats, the relationship between MT and nuchal muscle atonia, a widely recognized component of REM sleep. At all ages, muscle atonia preceded MT and persisted until awake behaviors occurred. In addition, muscle tone decreased gradually during transitions from awake behavior to twitching. Thus, MT during infancy occurs against a backdrop of muscle atonia, a result that is consistent with the view that AS is a developmental precursor of REM sleep.