Irina Topchiy

Local functional state differences between rat cortical columns.

Surface evoked potentials (SEPs) during auditory clicks and whisker twitches are usually larger during quiet sleep (QS) over waking and REM sleep. However, SEP amplitudes from single trials fluctuate periodically between high and low values regardless of sleep-wake cycle. To test the hypothesis that state-independent fluctuations represent local functional sleep-like states of individual cortical columns, we examined single trial SEP amplitudes from multiple cortical locations across sleep-wake cycles. Bilateral stimuli produced SEP amplitude fluctuations in each hemisphere that usually covaried (r = 0.4), but with frequent hemispheric differences. Two neighboring whiskers, twitched simultaneously on the same side, produced highly correlated SEPs in neighboring cortical columns (r = 0.9) with frequent divergences. We found 50% more disparity during QS over waking, indicating that the differences did not result from recording noise or stimulus inconsistency. Local SEP fluctuations also followed local differences in the delta wave signal during QS (r = 0.4), suggesting that similar mechanisms may modulate the SEP. The duration of the localized sleep-like (high SEP amplitude) state was dependent on the duration of prior wake-like (low SEP amplitude) state (r = 0.5), suggesting a use dependence of prior functional state period. Since SEP indicators fluctuated independently from whole animal sleep state, and were frequently different between hemispheres and nearby cortical columns, these data support the theory that sleep-like functional states may be localized to brain regions at least as small as cortical columns.

Cardiorespiratory effects of intertrigeminal area stimulation in vagotomized rats

It has been recently shown that the pontine intertrigeminal region (ITR) plays an important role in respiratory regulation, including vagally mediated apneic reflexes. Neurons of the ITR have connections with the nucleus tractus solitarius and projections to the ventrolateral medulla. However, the functional targets of these projections are not fully defined. Stimulation of ITR neurons produced respiratory effects, but cardiovascular responses have not been explored. We investigated impact of bilateral vagotomy on respiratory and cardiovascular responses to glutamate microinjections within the ITR in ketamine/xylazine anesthetized rats. Cardiorespiratory indices, including breath duration (TT), tidal volume (VT), mean cardiac intervals (RR), systolic blood pressure (SBP), pulse pressure (PP) and their coefficients of variation (CVTT, CVVT, CVSBP, CVPP, respectively) were analyzed in 30 s segments before and after injection of glutamate (10 mM, 30 L) into the ITR. This assessment was carried out both before and after bilateral vagotomy. Glutamate injection evoked apnea and increased CVTT, but these responses were not altered by bilateral vagotomy. In contrast, removing vagal pathways significantly increased volume variability (CVVT), making tidal volume more vulnerable to perturbation from the ITR. Vagotomy prolonged the increase of mean systolic blood pressure observed after glutamate injection and unmasked a delayed but sustained elevation of PP and CVPP after ITR stimulation. The present findings indicate a broad involvement of the ITR in autonomic regulation, including at least cardiovascular and respiratory effects.

Theta-rhythmic drive between medial septum and hippocampus in slow wave sleep and microarousal: A Granger causality analysis

Medial septum (MS) plays a critical role in controlling the electrical activity of the hippocampus (HIPP). In particular, theta-rhythmic burst firing of MS neurons is thought to drive lasting HIPP theta oscillations in rats during waking motor activity and REM sleep. Less is known about MS-HIPP interactions in nontheta states such as non-REM sleep, in which HIPP theta oscillations are absent but theta-rhythmic burst firing in subsets of MS neurons is preserved. The present study used Granger causality (GC) to examine the interaction patterns between MS and HIPP in slow-wave sleep (SWS, a nontheta state) and during its short interruptions called microarousals (a transient theta state). We found that during SWS, while GC revealed a unidirectional MS→HIPP influence over a wide frequency band (2-12 Hz, maximum: ∼8 Hz), there was no theta peak in the hippocampal power spectra, indicating a lack of theta activity in HIPP. In contrast, during microarousals, theta peaks were seen in both MS and HIPP power spectra and were accompanied by bidirectional GC with MS→HIPP and HIPP→MS theta drives being of equal magnitude. Thus GC in a nontheta state (SWS) vs. a theta state (microarousal) primarily differed in the level of HIPP→MS. The present findings suggest a modification of our understanding of the role of MS as the theta generator in two regards. First, a MS→HIPP theta drive does not necessarily induce theta field oscillations in the hippocampus, as found in SWS. Second, HIPP theta oscillations entail bidirectional theta-rhythmic interactions between MS and HIPP.

Functional topography of respiratory, cardiovascular and pontine-wave responses to glutamate microstimulation of the pedunculopontine tegmentum of the rat

Functionally distinct areas were mapped within the pedunculopontine tegmentum (PPT) of 42 ketamine/xylazine anesthetized rats using local stimulation by glutamate microinjection (10 mM, 5-12 nl). Functional responses were classified as: (1) apnea; (2) tachypnea; (3) hypertension (HTN); (4) sinus tachycardia; (5) genioglossus electromyogram activation or (6) pontine-waves (p-waves) activation.We found that short latency apneas were predominantly elicited by stimulation in the lateral portion of the PPT, in close proximity to cholinergic neurons. Tachypneic responses were elicited from ventral regions of the PPT and HTN predominated in the ventral portion of the antero-medial PPT. We observed sinus tachycardia after stimulation of the most ventral part of the medial PPT at the boundary with nucleus reticularis pontis oralis, whereas p-waves were registered predominantly following stimulation in the dorso-caudal portion of the PPT. Genioglossus EMG activation was evoked from the medial PPT. Our results support the existence of the functionally distinct areas within the PPT affecting respiration, cardiovascular function, EEG and genioglossus EMG.

Brown Norway and Zucker lean rats demonstrate circadian variation in ventilation and sleep apnea

STUDY OBJECTIVES Circadian rhythms influence many biological systems, but there is limited information about circadian and diurnal variation in sleep related breathing disorder. We examined circadian and diurnal patterns in sleep apnea and ventilatory patterns in two rat strains, one with high sleep apnea propensity (Brown Norway [BN]) and the other with low sleep apnea propensity (Zucker Lean [ZL]). DESIGN/SETTING Chronically instrumented rats were randomized to breathe room air (control) or 100% oxygen (hyperoxia), and we performed 20-h polysomnography beginning at Zeitgeber time 4 (ZT 4; ZT 0 = lights on, ZT12 = lights off). We examined the effect of strain and inspired gas (twoway analysis of variance) and analyzed circadian and diurnal variability. MEASUREMENTS AND RESULTS Strain and inspired gas-dependent differences in apnea index (AI; apneas/h) were particularly prominent during the light phase. AI in BN rats (control, 16.9 ± 0.9; hyperoxia, 34.0 ± 5.8) was greater than in ZL rats (control, 8.5 ± 1.0; hyperoxia, 15.4 ± 1.1, [strain effect, P < 0.001; gas effect, P = 0.001]). Hyperoxia reduced respiratory frequency in both strains, and all respiratory pattern variables demonstrated circadian variability. BN rats exposed to hyperoxia demonstrated the largest circadian fluctuation in AI (amplitude = 17.9 ± 3.7 apneas/h [strain effect, P = 0.01; gas effect, P < 0.001; interaction, P = 0.02]; acrophase = 13.9 ± 0.7 h; r (2) = 0.8 ± 1.4). CONCLUSIONS Inherited, environmental, and circadian factors all are important elements of underlying sleep related breathing disorder. Our method to examine sleep related breathing disorder phenotypes in rats may have implications for understanding vulnerability for sleep related breathing disorder in humans.

Acute exacerbation of sleep apnea by hyperoxia impairs cognitive flexibility in Brown-Norway rats.

STUDY OBJECTIVES To determine whether learning deficits occur during acute exacerbation of spontaneous sleep related breathing disorder (SRBD) in rats with high (Brown Norway; BN) and low (Zucker Lean; ZL) apnea propensity. DESIGN Spatial acquisition (3 days) and reversal learning (3 days) in the Morris water maze (MWM) with polysomnography (12:00-08:00): (1) with acute SRBD exacerbation (by 20-h hyperoxia immediately preceding reversal learning) or (2) without SRBD exacerbation (room air throughout). SETTING Randomized, placebo-controlled, repeated-measures design. PARTICIPANTS 14 BN rats; 16 ZL rats. INTERVENTIONS 20-h hyperoxia. MEASUREMENTS AND RESULTS Apneas were detected as cessation of respiration ≥ 2 sec. Swim latency in MWM, apnea indices (AI; apneas/hour of sleep) and percentages of recording time for nonrapid eye movement (NREM), rapid eye movement (REM), and total sleep were assessed. Baseline AI in BN rats was more than double that of ZL rats (22.46 ± 2.27 versus 10.7 ± 0.9, P = 0.005). Hyperoxia increased AI in both BN (34.3 ± 7.4 versus 22.46 ± 2.27) and ZL rats (15.4 ± 2.7 versus 10.7 ± 0.9) without changes in sleep stage percentages. Control (room air) BN and ZL rats exhibited equivalent acquisition and reversal learning. Acute exacerbation of AI by hyperoxia produced a reversal learning performance deficit in BN but not ZL rats. In addition, the percentage of REM sleep and REM apnea index in BN rats during hyperoxia negatively correlated with reversal learning performance. CONCLUSIONS Acute exacerbation of sleep related breathing disorder by hyperoxia impairs reversal learning in a rat strain with high apnea propensity, but not a strain with a low apnea propensity. This suggests a non-linear threshold effect may contribute to the relationships between sleep apnea and cognitive dysfunctions, but strain-specific differences also may be important.

Reciprocal Interactions between Medial Septum and Hippocampus in Theta Generation: Granger Causality Decomposition of Mixed Spike-Field Recordings

The medial septum (MS) plays an essential role in rhythmogenesis in the hippocampus (HIPP); theta-rhythmic bursts of MS neurons are believed to drive theta oscillations in rats’ HIPP. The MS theta pacemaker hypothesis has solid foundation but the MS-hippocampal interactions during different behavioral states are poorly understood. The MS and the HIPP have reciprocal connections and it is not clear in particular what role, if any, the strong HIPP to MS projection plays in theta generation. To study the functional interactions between MS and HIPP during different behavioral states, this study investigated the relationship between MS single-unit activity and HIPP field potential oscillations during theta states of active waking and REM sleep and non-theta states of slow wave sleep (SWS) and quiet waking (QW), i.e., sleep-wake states that comprise the full behavioral repertoire of undisturbed, freely moving rats. We used non-parametric Granger causality (GC) to decompose the MS-HIPP synchrony into its directional components, MS→HIPP and HIPP→MS, and to examine the causal interactions between them within the theta frequency band. We found a significant unidirectional MS→HIPP influence in non-theta states which switches to bidirectional theta drive during theta states with MS→HIPP and HIPP→MS GC being of equal magnitude. In non-theta states, unidirectional MS→HIPP influence was accompanied by significant MS-HIPP coherence, but no signs of theta oscillations in the HIPP. In theta states of active waking and REM sleep, sharp theta coherence and strong theta power in both structures was associated with a rise in HIPP→MS to the level of the MS→HIPP drive. Thus, striking differences between waking and REM sleep theta states and non-theta states of SWS and QW were primarily observed in activation of theta influence carried by the descending HIPP→MS pathway associated with more regular rhythmic bursts in the MS and sharper MS→HIPP GC spectra without a significant increase in MS→HIPP GC magnitude. The results of this study suggest an essential role of descending HIPP to MS projections in theta generation.

Impact of the vagal feedback on cardiorespiratory coupling in anesthetized rats

Cardiorespiratory coupling can be significantly influenced by both pontine and vagal modulation of medullary motor and premotor areas. We investigated influences of the pontine intertrigeminal region (ITR) and peripheral vagal pathways on the coupling between systolic blood pressure (SBP) and respiration in 9 anesthetized rats. Glutamate injection into the ITR perturbed both respiration and SBP and decreased SBP-respiratory coherence (0.95±0.01 vs 0.89±0.02; (p=0.01). Intravenous infusion of serotonin (5-HT) produced apnea and hypertension and also decreased SBP-respiratory coherence (0.95±0.01 vs 0.72±0.06; p=0.04). Bilateral vagotomy eliminated the cardiorespiratory coherence perturbations induced by central (glutamate injection into the ITR: 0.89±0.03 vs 0.86±0.03; p=0.63) and peripheral (5-HT infusion: 0.89±0.03 vs 0.88±0.02; p=0.98) pharmacologic manipulations. Glutamate stimulation of the ITR postvagotomy increased the relative spectral power density of SBP in the respiratory frequency range (0.25±0.08 vs 0.55±0.06; p=0.01). The data suggest that SBP-respiratory coupling is largely mediated within the central nervous system, with vagal systems acting in a way that disrupts coherence during transient cardiorespiratory disturbances. Although decreased cardiorespiratory coherence may increase cardiac work during perturbations, this may be physiologically advantageous in restoring homeostatic equilibrium of respiration and blood pressure.