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Biomedical subjects

David M Rector

Publications and source records attributed to David M Rector.

13 recordsLinked to original sources

Hemispheric mapping of secondary somatosensory cortex in the rat.

This study used high-resolution hemispheric mapping of somatosensory evoked potentials to determine the number and organization of secondary somatosensory areas (SII) in rat cortex. Two areas, referred to as SII and PV (parietoventral), revealed complete (SII) or nearly complete (PV) body maps. The vibrissa and somatic representation of SII was upright, rostrally oriented, and immediately lateral to primary somatosensory cortex (SI), with a dominant face representation. Vibrissa representations in SII were highly organized, with the rows staggered rostrally along the mediolateral axis. Area PV was approximately one fifth the size of SII, and located rostral and lateral to auditory cortex. PV had a rostrally oriented and inverted body representation that was dominated by the distal extremities, with little representation of the face or vibrissae. These data support the conclusion that in the rat, as in other species, SII and PV represent anatomically and functionally distinct areas of secondary somatosensory cortex.

Afferent Pathways↗

Evoked response potential markers for anesthetic and behavioral states.

The rodent whisker sensory system is a commonly used model of cortical processing; however, anesthetics cause profound differences in the shape and timing of evoked responses. Evoked response studies, especially those that use spatial mapping techniques, such as fMRI or optical imaging, will thus show significantly different results depending on the anesthesia used. To describe the effect of behavioral states and commonly used anesthetics, we characterized the early surface-evoked response potentials (ERPs) components (first ERP peak: gamma band 25-45 Hz; fast oscillation: 200-400 Hz; and very fast oscillation: 400-600 Hz) using a 25-channel electrode array on the somatosensory cortex during whisker stimulation. We found significant differences in the ERP shape when ketamine/xylazine, urethane, propofol, isoflurane, and pentobarbital sodium were administered and during sleep and wake states. The highest ERP amplitudes were observed under propofol anesthesia and during quiet sleep. Under isoflurane, the ERP was nearly absent, except for a very late component, which was concombinant with burst synchronization. The slowest responses were seen under urethane and propofol anesthesia. Spatial mapping experiments that use electrical, NMR, or optical techniques must consider the anesthetic dependency of these signals, especially when stimulation protocols or electrical and metabolic responses are compared.

Anesthesia, General↗

A MEMS fabricated flexible electrode array for recording surface field potentials.

We developed a method to microfabricate flexible electrode arrays on a thin Kapton substrate, which was engineered to minimize trauma when inserted between the dura and skull to obtain surface EEG recordings. The array consisted of 64 gold electrodes, each 150microm in diameter on a 750microm spaced 8x8 grid. Using photolithographic procedures, any arrangement of electrodes can be implemented. We used the electrode array to record evoked response signals to create topographical maps of the whisker barrels on the cortical surface with excellent signal stability over a period of 8h. The materials used for this fabrication are potentially biologically inert and, with some additional modifications to the design, can be chronically implanted with minimal side effects. Retinal prosthesis, human neurosurgery, and neurological research are all limited to some degree by the resolution and biological compatibility of the implants used. This type of array could greatly enhance the spatial resolution, signal quality, and stability of implantable surface electrode arrays.

Animals↗

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.

Acoustic Stimulation↗

Optimized birefringence changes during isolated nerve activation.

Single trial, birefringence signals associated with action potentials from isolated lobster nerves were optimized with high-intensity light-emitting diodes (LEDs) and glass polarizers. The narrow spectral output of the LEDs allowed us to select specific wavelengths, increasing the effectiveness of the polarizers and minimizing the stray light in the system. The LEDs produced intensity profiles equivalent to narrowband filtered 100-W halogen light, and birefringence signals were comparable or superior in size and clarity to halogen lamp recordings. The results support a direct correlation between signal size and polarizer extinction coefficient. Increasing the sensitivity of birefringence detection through the use of LED light sources could ameliorate noninvasive brain imaging techniques that employ fast optical consequences associated with action potential propagation.

Action Potentials↗

Spatio-temporal mapping of rat whisker barrels with fast scattered light signals.

Optical techniques offer a number of potential advantages for imaging dynamic spatio-temporal patterns of activity in neural tissue. The methods provide the wide field of view required to image population activation across networks, while allowing resolution of the detailed structure of individual cells. Optical probes can provide high temporal resolution without penetrating the tissue surface. However, functional optical imaging has been constrained by the small size of the signals and the sluggish nature of the metabolic and hemodynamic responses that are the basis of most existing methods. Here, we employ both high-speed CCD cameras and high-sensitivity photodiodes to optimize resolution in both space and time, together with dark-field illumination in the near-infrared, to record fast intrinsic scattering signals from rat somatosensory cortex in vivo. Optical responses tracked the physiological activation of cortical columns elicited by single whisker twitches. High-speed imaging produced maps that were initially restricted in space to individual barrels, and then spread over time. Photodiode recordings disclosed 400-600 Hz oscillatory responses, tightly correlated in frequency and phase to those seen in simultaneous electrical recordings. Imaging based on fast intrinsic light scattering signals eventually could provide high resolution dynamic movies of neural networks in action.

Animals↗

Cross-polarized reflected light measurement of fast optical responses associated with neural activation.

We developed an optical probe for cross-polarized reflected light measurements and investigated optical signals associated with electrophysiological activation in isolated lobster nerves. The cross-polarized baseline light intensity (structural signal) and the amplitude of the transient response to stimulation (functional signal) measured in reflected mode were dependent on the orientation of the nerve axis relative to the polarization plane of incident light. The maximum structural signal and functional response amplitude were observed at 45 degrees , and the ratio of functional to structural signals was approximately constant across orientations. Functional responses were measured in single trials in both transmitted and reflected geometries and responses had similar waveforms. Both structural and functional signals were an order of magnitude smaller in reflected than in transmitted light measurements, but functional responses had similar signal/noise ratios. We propose a theoretical model based on geometrical optics that is consistent with experimental results. In the model, the cross-polarized structural signal results from light reflection from axonal fibers and the transient functional response arises from axonal swelling associated with neural activation. Polarization-sensitive reflected light measurements could greatly enhance in vivo imaging of neural activation since cross-polarized responses are much larger than scattering signals now employed for dynamic functional neuroimaging.

Animals↗

Simultaneous birefringence and scattered light measurements reveal anatomical features in isolated crustacean nerve.

Simultaneous fast birefringence and scattered light changes associated with crustacean nerve activation have different time courses and are produced by separate biophysical mechanisms. Technological advances in illumination, photodiodes and amplification circuitry achieved better signal-to-noise than earlier studies revealing optical signals in axonal nerve bundles as small as crayfish ventral cord and claw. The birefringence measurements yielded signals that could be observed in single trials, with temporally separated peaks associated with axonal populations of different diameters. A slit aperture placed perpendicular to the nerve reduced the spatial-temporal integration and enhanced the temporal structure of the separate peaks in the birefringence signal. Moving the slit aperture farther from the stimulation point delayed the signal in time, and also enhanced the separation between peaks. Different propagation velocities of the separate peaks provided evidence for at least three neuronal populations in the bundle. These studies underscore the advantages of birefringence over scattering measurements. Application of birefringence methods can optimize non-invasive imaging techniques being developed to detect fast optical responses associated with electrical neural activity in humans.

Animals↗

Activity changes of the cat paraventricular hypothalamus during stressor exposure.

Dorso-medial paraventricular hypothalamus (PVH) activity was assessed by light scattering procedures in freely behaving cats during auditory stressor exposure. Acoustic noise (> 95dB) raised plasma ACTH concentrations, somatic muscle tonus, respiratory frequency and cardiac rates; PVH activity peaked 0.8s following stimulation, and then markedly declined below baseline to a trough at 9.7s. Hypothalamic responses were not uniformly distributed across the recorded PVH field. Activity changes emerged from subregions within the visualized area, and were widespread at the overall activity zenith and nadir. Isolated pixels appeared opposite in activity pattern to overall changes. We suggest that transient activity increases represent initial PVH neural stress responses, and that subsequent profound declines result from neural inhibitory feedback.

Acoustic Stimulation↗

Late-developing rostral ventrolateral medullary surface responses to cardiovascular challenges during sleep.

Pressor and depressor manipulations are usually followed by compensatory autonomic, respiratory, somatomotor or arousal responses that limit the extent of blood pressure change. Of neural sites participating in blood pressure control, the rostral ventrolateral medullary surface (RVLMS) contributes significantly, and exhibits rapid-onset overall activity declines and increases to pressor and depressor challenges, respectively. In addition, longer-latency physiological responses develop that further compensate for the homeostatic challenge; some of these later influences are associated with arousal. Late-developing RVLMS activity changes accompanying physiologic responses that normalize a cardiovascular manipulation may provide insights into compensatory neural mechanisms during sleep following sustained or extreme blood pressure changes. We used intrinsic optical imaging procedures in seven unanesthetized adult cats to examine RVLMS and control site responses to pressor and depressor challenges during sleep that resulted in somatomotor, respiratory, heart rate or electroencephalographic indications of late-developing (post-baroreflex) compensatory responses. Although initial RVLMS responses differed in direction between pressor and depressor challenges, neural activity increased later in both manipulations, coincident with overt physiological manifestations indicative of compensatory responses, including arousal. Arousal occurred in 44% of blood pressure challenges. Comparable late-developing neural activity increases were not apparent in control sites. Latencies of late RVLMS responses during rapid eye movement sleep were significantly longer than in quiet sleep for pressor challenges. The pattern of the late RVLMS responses was not dependent on arousal, and suggests that the RVLMS participates in both the early baroreflex response and the late-developing compensatory actions.

Animals↗

Optical lever recording of displacements from activated lobster nerve bundles and Nitella internodes.

An optical lever was designed for studying physical displacements associated with electrophysiological activation of lobster nerve bundles. Stimulation current pulses generated a compound action potential volley, and upward physical displacements of <1 nm were recorded. The swelling displacement propagated in the same direction as the action potential volley, occurred simultaneously with the action potentials, and required 10 ms to relax after the electrical potential was restored. For comparison with previous reports, we also recorded the displacement of Nitella internodes associated with electrical stimulation. We found that a rapid swelling displacement (approximately 10 nm) was followed by a larger, slow-shrinking displacement (approximately 100 nm).

Animals↗

State-dependent columnar organization of dorsal hippocampal activity in the freely-behaving cat.

We examined reflected light images from the dorsal hippocampus in freely-behaving cats to assess the topographical organization of neural activity expressed during sleep-waking states. Reflectance patterns, organized as columns along the CA3-CA1 axis which appeared during rapid eye movement sleep (REM) and waking states, were compared to baseline periods collected during quiet sleep (QS). The columns were not static across states or behaviors within a state, but showed inversion in activation/inactivation patterns, and narrowed or widened over time. A correlation between state-dependence and reflectance patterns emerged, with transitions to active waking and REM most likely exhibiting enhanced columnar patterning, relative to QS. No columnar patterns appeared in overlying neocortical areas or in control media. The columnar patterns suggest a highly structured functional state-dependent organization in the dorsal hippocampus, as has been previously predicted by anatomical studies.

Animals↗

High-performance confocal system for microscopic or endoscopic applications.

We designed a high-performance confocal system that can be easily adapted to an existing light microscope or coupled with an endoscope for remote imaging. The system employs spatially and temporally patterned illumination produced by one of several mechanisms, including a micromirror array video projection device driven by a computer video source or a microlens array scanned by a piezo actuator in the microscope illumination path. A series of subsampled "component" video images are acquired from a solid-state video camera. Confocal images are digitally reconstructed using "virtual pinhole" synthetic aperture techniques applied to the collection of component images. Unlike conventional confocal techniques that raster scan a single detector and illumination point, our system samples multiple locations in parallel, with particular advantages for monitoring fast dynamic processes. We compared methods of patterned illumination and confocal image reconstruction by characterizing the point spread function, contrast, and intensity of imaged objects. Sample 3D reconstructions include a diatom and a Golgi-stained nerve cell collected in transmission.

Animals↗