Cardiorespiratory rhythm-contingent trace eyeblink conditioning in elderly adults.

Learning outcome is modified by the degree to which the subject responds and pays attention to specific stimuli. Our recent research suggests that presenting stimuli in contingency with a specific phase of the cardiorespiratory rhythm might expedite learning. Specifically, expiration-diastole (EXP-DIA) is beneficial for learning trace eyeblink conditioning (TEBC) compared with inspiration-systole (INS-SYS) in healthy young adults. The aim of this study was to investigate whether the same holds true in healthy elderly adults (n = 50, aged >70 yr). Participants were instructed to watch a silent nature film while TEBC trials were presented at either INS-SYS or EXP-DIA (separate groups). Learned responses were determined as eyeblinks occurring after the tone conditioned stimulus (CS), immediately preceding the air puff unconditioned stimulus (US). Participants were classified as learners if they made at least five conditioned responses (CRs). Brain responses to the stimuli were measured by electroencephalogram (EEG). Memory for the film and awareness of the CS-US contingency were evaluated with a questionnaire. As a result, participants showed robust brain responses to the CS, acquired CRs, and reported awareness of the CS-US relationship to a variable degree. There was no difference between the INS-SYS and EXP-DIA groups in any of the above. However, when only participants who learned were considered, those trained at EXP-DIA (n = 11) made more CRs than those trained at INS-SYS (n = 13). Thus, learned performance could be facilitated in those elderly who learned. However, training at a specific phase of cardiorespiratory rhythm did not increase the proportion of participants who learned.NEW & NOTEWORTHY We trained healthy elderly individuals in trace eyeblink conditioning, either at inspiration-systole or at expiration-diastole. Those who learned exhibited more conditioned responses when trained at expiration-diastole rather than inspiration-systole. However, there was no difference between the experimental groups in the proportion of individuals who learned or did not learn.

CA3-CA1 long-term potentiation occurs regardless of respiration and cardiac cycle phases in urethane-anesthetized rats.

Breathing and heartbeat synchronize to each other and to brain function and affect cognition in humans. However, it is not clear how cardiorespiratory rhythms modulate such basic processes as synaptic plasticity thought to underlie learning. Thus, we studied if respiration and cardiac cycle phases at burst stimulation onset affect hippocampal long-term potentiation (LTP) in the CA3-CA1 synapse in urethane-anesthetized adult male Sprague-Dawley rats. In a between-subjects design, we timed burst stimulation of the ventral hippocampal commissure (vHC) to systole or diastole either during expiration or inspiration and recorded responses throughout the hippocampus with a linear probe. As classical conditioning in humans seems to be most efficient at expiration-diastole, we also expected LTP to be most efficient if burst stimulation was targeted to expiration-diastole. However, LTP was induced equally in all four groups and respiration and cardiac cycle phase did not modulate CA1 responses to vHC stimulation overall. This could be perhaps because we bypassed all natural routes of external influences on the CA1 by directly stimulating the vHC. In the future, the effect of cardiorespiratory rhythms on synaptic plasticity could also be studied in awake state and in other parts of the hippocampal tri-synaptic loop.

Hippocampal responses to electrical stimulation of the major input pathways are modulated by dentate spikes.

Dentate gyrus (DG) is important for pattern separation and spatial memory, and it is thought to gate information flow to the downstream hippocampal subregions. Dentate spikes (DSs) are high-amplitude, fast, positive local-field potential events taking place in the DG during immobility and sleep, and they have been connected to memory consolidation in rodents. DSs are a result of signaling from the entorhinal cortex (EC) to the DG, and they suppress firing of pyramidal cells in the CA3 and CA1. To study the effects of DSs to signaling in the hippocampal tri-synaptic loop, we electrically stimulated the afferent fibers of the DG, CA3, and CA1 in adult male Sprague-Dawley rats at different delays from DSs. Responses to stimulation were increased in the EC-DG synapse during DSs, and the effect was amplified after theta-burst stimulation. We concluded that DSs strengthen the excitatory signal from the EC to the DG, which is reinforced by synapse potentiation and increased excitability of granule cells after theta-burst stimulation. This signal boosting may function in enhancing plastic changes in the DG-CA3 synapse. As responses in the CA3 and CA1 remained unaffected by the DS, the DS-contingent silencing of pyramidal cells seems to be a result of a decrease in excitatory input rather than a decrease in the excitability of the pyramidal cells themselves. In addition, we found that the DSs occur asynchronously in the left and right hippocampi, giving novel evidence of lateralization of the rodent hippocampus.

Cardiac cycle and respiration phase affect responses to the conditioned stimulus in young adults trained in trace eyeblink conditioning.

Rhythms of breathing and heartbeat are linked to each other as well as to the rhythms of the brain. Our recent studies suggest that presenting conditioned stimulus during expiration or during the diastolic phase of the cardiac cycle facilitates neural processing of that stimulus and improves learning in a conditioning task. To date, it has not been examined whether using information from both respiration and cardiac cycle phases simultaneously allows even more efficient modulation of learning. Here, we studied whether the timing of the conditioned stimulus to different cardiorespiratory rhythm phase combinations affects learning in a conditioning task in healthy young adults. The results were consistent with previous reports: timing the conditioned stimulus to diastole during expiration was more beneficial for learning than timing it to systole during inspiration. Cardiac cycle phase seemed to explain most of this variation in learning at the behavioral level. Brain-evoked potentials (N1) elicited by the conditioned stimulus and recorded using electroencephalogram were larger when the conditioned stimulus was presented to diastole during expiration than when it was presented to systole during inspiration. Breathing phase explained the variation in the N1 amplitude. To conclude, our findings suggest that noninvasive monitoring of bodily rhythms combined with closed-loop control of stimulation can be used to promote learning in humans. The next step will be to test if performance can also be improved in humans with compromised cognitive ability, such as in older people with memory impairments.NEW & NOTEWORTHY We report, for the first time, that the rhythms of breathing and the beating of the heart have a phase combination that is indicative of a neural state beneficial for cognition. This suggests that bodily rhythms not only modulate cognition but that this phenomenon can also be noninvasively harnessed to improve learning in humans.

Irradiation of the head reduces adult hippocampal neurogenesis and impairs spatial memory, but leaves overall health intact in rats.

Treatment of brain cancer, glioma, can cause cognitive impairment as a side-effect, possibly because it disrupts the integrity of the hippocampus, a structure vital for normal memory. Radiotherapy is commonly used to treat glioma, but the effects of irradiation on the brain are still poorly understood, and other biological effects have not been extensively studied. Here, we exposed healthy adult male rats to moderate-dose irradiation of the head. We found no effect of irradiation on systemic inflammation, weight gain or gut microbiota diversity, although it increased the abundance of Bacteroidaceae family, namely Bacteroides genus in the gut microbiota. Irradiation had no effect on long-term potentiation in the CA3-CA1 synapse or endogenous hippocampal electrophysiology, but it did reduce adult hippocampal neurogenesis and impaired short-term spatial recognition memory. However, no overall cognitive impairment was observed. To summarize, our results suggest that in adult male rats head irradiation does not compromise health or cognition overall even though the number of new, adult-born hippocampal neurons is decreased. Thus, the sole effects of head irradiation on the body, brain and cognition might be less harmful than previously thought, and the cognitive decline experienced by cancer patients might originate from physiological and mental effects of the disease itself. Therefore, to increase the translational value of animal studies, the effects of irradiation should be studied together with cancer, in older animals, using varying irradiation protocols and doses.

Most hippocampal CA1 pyramidal cells in rabbits increase firing during awake sharp-wave ripples and some do so in response to external stimulation and theta.

Hippocampus forms neural representations of real-life events including multimodal information of spatial and temporal context. These representations, i.e., organized sequences of neuronal firing, are repeated during following rest and sleep, especially when so-called sharp-wave ripples (SPW-Rs) characterize hippocampal local field potentials. This SPW-R -related replay is thought to underlie memory consolidation. Here, we set out to explore how hippocampal CA1 pyramidal cells respond to the conditioned stimulus during trace eyeblink conditioning and how these responses manifest during SPW-Rs in awake adult female New Zealand White rabbits. Based on reports in rodents, we expected SPW-Rs to take place in bursts, possibly according to a slow endogenous rhythm. In awake rabbits, half of all SPW-Rs took place in bursts, but no endogenous slow rhythm appeared. Conditioning trials suppressed SPW-Rs while increasing theta for a period of several seconds. As expected based on previous findings, only a quarter of the putative CA1 pyramidal cells increased firing in response to the conditioned stimulus. Compared with other cells, rate-increasing cells were more active during spontaneous epochs of hippocampal theta while response profile during conditioning did not affect firing during SPW-Rs. Taken together, CA1 pyramidal cell firing during SPW-Rs is not limited to cells that fired during the preceding experience. Furthermore, the importance of possible reactivations taking place during theta epochs on memory consolidation warrants further investigation.NEW & NOTEWORTHY We studied hippocampal sharp-wave ripples and theta and CA1 pyramidal cell activity during trace eyeblink conditioning in rabbits. Conditioning trials suppressed ripples while increasing theta for a period of several seconds. A quarter of the cells increased firing in response to the conditioned stimulus and fired extensively during endogenous theta as well as ripples. The role of endogenous theta epochs in off-line memory consolidation should be studied further.

Breathe out and learn: Expiration-contingent stimulus presentation facilitates associative learning in trace eyeblink conditioning.

Rhythmic variation in heart rate and respiratory pattern are coupled in a way that optimizes the level of oxygen in the blood stream of the lungs and the body as well as saves energy in pulmonary gas exchange. It has been suggested that the cardiac cycle and respiratory pattern are coupled to neural oscillations of the brain. Yet, studies on how this rhythmic coupling is related to behavior are scarce. There is some evidence that, for example, the phase of respiration affects memory retrieval and the electrophysiological oscillatory state of the limbic system. It is also known that the phase of the cardiac cycle and hippocampal electrophysiological oscillations alone affect learning. Here, we studied whether the timing of training trials to different phases of respiration affects learning trace eyeblink conditioning in healthy adult humans. Trials consisting of a neutral conditioned stimulus (200-ms tone) and a slightly aversive unconditioned stimulus (100-ms air puff toward the eye), presented with a 600-ms trace interval, were timed to either inspiration or expiration. A control group was trained regardless of respiratory phase. We found that, at the end of training, the rate of conditioned responses was higher in the group trained at expiration than it was in the other two groups. That is, brain state seems to fluctuate as a function of respiratory rhythm, and this fluctuation is also behaviorally relevant, exerting its effect on, at the least, a simple form of associative learning.

Dentate spikes and learning: disrupting hippocampal function during memory consolidation can improve pattern separation.

Hippocampal dentate spikes (DSs) are short-duration, large-amplitude fluctuations in hilar local field potentials and take place while resting and sleeping. During DSs, dentate gyrus granule cells increase firing while CA1 pyramidal cells decrease firing. Recent findings suggest DSs play a significant role in memory consolidation after training on a hippocampus-dependent, nonspatial associative learning task. Here, we aimed to find out whether DSs are important in other types of hippocampus-dependent learning tasks as well. To this end, we trained adult male Sprague-Dawley rats in a spatial reference memory task, a fixed interval task, and a pattern separation task. During a rest period immediately after each training session, we either let neural activity to take place as usual, timed electrical stimulation of the ventral hippocampal commissure (vHC) to immediately follow DSs, or applied the vHC stimulation during a random neural state. We found no effect of vHC stimulation on performance in the spatial reference memory task or in the fixed interval task. Surprisingly, vHC stimulation, especially contingent on DSs, improved performance in the pattern separation task. In conclusion, the behavioral relevance of hippocampal processing and DSs seems to depend on the task at hand. It could be that in an intact brain, offline memory consolidation by default involves associating neural representations of temporally separate but related events. In some cases this might be beneficial for adaptive behavior in the future (associative learning), while in other cases it might not (pattern separation). NEW & NOTEWORTHY The behavioral relevance of dentate spikes seems to depend on the learning task at hand. We suggest that dentate spikes are related to associating neural representations of temporally separate but related events within the dentate gyrus. In some cases this might be beneficial for adaptive behavior in the future (associative learning), while in other cases it might not (pattern separation).

Learning by heart: cardiac cycle reveals an effective time window for learning.

Cardiac cycle phase is known to modulate processing of simple sensory information. This effect of the heartbeat on brain function is likely exerted via baroreceptors, the neurons sensitive for changes in blood pressure. From baroreceptors, the signal is conveyed all the way to the forebrain and the medial prefrontal cortex. In the two experiments reported, we examined whether learning, as a more complex form of cognition, can be modulated by the cardiac cycle phase. Human participants ( experiment 1) and rabbits ( experiment 2) were trained in trace eyeblink conditioning while neural activity was recorded. The conditioned stimulus was presented contingently with either the systolic or diastolic phase of the cycle. The tone used as the conditioned stimulus evoked amplified responses in both humans (electroencephalogram from "vertex," Cz) and rabbits (hippocampal CA1 local field potential) when its onset was timed at systole. In humans, the cardiac cycle phase did not affect learning, but rabbits trained at diastole learned significantly better than those trained at a random phase of the cardiac cycle. In summary, our results suggest that neural processing of external stimuli and also learning can be affected by targeting stimuli on the basis of cardiac cycle phase. These findings might be useful in applications aimed at maximizing or minimizing the effects of external stimulation. NEW & NOTEWORTHY It has been shown that rapid changes in bodily states modulate neural processing of external stimulus in brain. In this study, we show that modulation of neural processing of external stimulus and learning about it depends on the phase of the cardiac cycle. This is a novel finding that can be applied to optimize associative learning.

Hippocampal theta phase-contingent memory retrieval in delay and trace eyeblink conditioning.

Hippocampal theta oscillations (3-12Hz) play a prominent role in learning. It has been suggested that encoding and retrieval of memories are supported by different phases of the theta cycle. Our previous study on trace eyeblink conditioning in rabbits suggests that the timing of the conditioned stimulus (CS) in relation to theta phase affects encoding but not retrieval of the memory trace. Here, we directly tested the effects of hippocampal theta phase on memory retrieval in two experiments conducted on adult female New Zealand White rabbits. In Experiment 1, animals were trained in trace eyeblink conditioning followed by extinction, and memory retrieval was tested by presenting the CS at troughs and peaks of the theta cycle during different stages of learning. In Experiment 2, animals were trained in delay conditioning either contingent on a high level of theta or at a random neural state. Conditioning was then followed by extinction conducted either at a random state, contingent on theta trough or contingent on theta peak. Our current results indicate that the phase of theta at CS onset has no effect on the performance of the behavioral learned response at any stage of classical eyeblink conditioning or extinction. In addition, theta-contingent trial presentation does not improve learning during delay eyeblink conditioning. The results are consistent with our earlier findings and suggest that the theta phase alone is not sufficient to affect learning at the behavioral level. It seems that the retrieval of recently acquired memories and consequently performing a learned response is moderated by neural mechanisms other than hippocampal theta.

Automatic auditory and somatosensory brain responses in relation to cognitive abilities and physical fitness in older adults.

In normal ageing, structural and functional changes in the brain lead to an altered processing of sensory stimuli and to changes in cognitive functions. The link between changes in sensory processing and cognition is not well understood, but physical fitness is suggested to be beneficial for both. We recorded event-related potentials to somatosensory and auditory stimuli in a passive change detection paradigm from 81 older and 38 young women and investigated their associations with cognitive performance. In older adults also associations to physical fitness were studied. The somatosensory mismatch response was attenuated in older adults and it associated with executive functions. Somatosensory P3a did not show group differences, but in older adults, it associated with physical fitness. Auditory N1 and P2 responses to repetitive stimuli were larger in amplitude in older than in young adults. There were no group differences in the auditory mismatch negativity, but it associated with working memory capacity in young but not in older adults. Our results indicate that in ageing, changes in stimulus encoding and deviance detection are observable in electrophysiological responses to task-irrelevant somatosensory and auditory stimuli, and the higher somatosensory response amplitudes are associated with better executive functions and physical fitness.

Hippocampal electrical stimulation disrupts associative learning when targeted at dentate spikes.

KEY POINTS: Dentate spikes are fast fluctuations of hilar local-field potentials that take place during rest and are thought to reflect input arriving from the entorhinal cortex to the hippocampus. During dentate spikes, neuronal firing in hippocampal input (dentate gyrus) and output (CA1/CA3) regions is uncoupled. To date, the behavioural significance of dentate spikes is unknown. Here, we provide evidence that disrupting the dentate spike-related uncoupling of the dentate gyrus and the CA1/CA3 subregions for 1 h after training retards associative learning. We suggest dentate spikes play a significant role in memory consolidation. ABSTRACT: Hippocampal electrophysiological oscillations, namely theta and ripples, have been implicated in encoding and consolidation of new memories, respectively. According to existing literature, hippocampal dentate spikes are prominent, short-duration (<30 ms), large-amplitude (∼2-4 mV) fluctuations in hilar local-field potentials that take place during awake immobility and sleep. Interestingly, previous studies indicate that during dentate spikes dentate gyrus granule cells increase their firing while firing of CA1 pyramidal cells are suppressed, thus resulting in momentary uncoupling of the two hippocampal subregions. To date, the behavioural significance of dentate spikes is unknown. Here, to study the possible role of dentate spikes in learning, we trained adult male Sprague-Dawley rats in trace eyeblink classical conditioning. For 1 h immediately following each conditioning session, one group of animals received hippocampal stimulation via the ventral hippocampal commissure (vHC) contingent on dentate spikes to disrupt the uncoupling between the dentate gyrus and the CA1 subregions. A yoked control group was stimulated during immobility, irrespective of brain state, and another control group was not stimulated at all. As a result, learning was impaired only in the group where vHC stimulation was administered contingent on dentate spikes. Our results suggest dentate spikes and/or the associated uncoupling of the dentate gyrus and the CA1 play a significant role in memory consolidation. Dentate spikes could possibly reflect reactivation and refinement of a memory trace within the dentate gyrus triggered by input from the entorhinal cortex.

Phase matters: responding to and learning about peripheral stimuli depends on hippocampal θ phase at stimulus onset.

Hippocampal θ (3-12 Hz) oscillations are implicated in learning and memory, but their functional role remains unclear. We studied the effect of the phase of local θ oscillation on hippocampal responses to a neutral conditioned stimulus (CS) and subsequent learning of classical trace eyeblink conditioning in adult rabbits. High-amplitude, regular hippocampal θ-band responses (that predict good learning) were elicited by the CS when it was timed to commence at the fissure θ trough (Trough group). Regardless, learning in this group was not enhanced compared with a yoked control group, possibly due to a ceiling effect. However, when the CS was consistently presented to the peak of θ (Peak group), hippocampal θ-band responding was less organized and learning was retarded. In well-trained animals, the hippocampal θ phase at CS onset no longer affected performance of the learned response, suggesting a time-limited role for hippocampal processing in learning. To our knowledge, this is the first study to demonstrate that timing a peripheral stimulus to a specific phase of the hippocampal θ cycle produces robust effects on the synchronization of neural responses and affects learning at the behavioral level. Our results support the notion that the phase of spontaneous hippocampal θ oscillation is a means of regulating the processing of information in the brain to a behaviorally relevant degree.