Faculty Bibliography

A growing number of minimally invasive surgical and diagnostic procedures require the insertion of an optical, mechanical, or electronic device in narrow spaces inside a human body. In such procedures, precise motion control is essential to avoid damage to the patient's tissues and/or the device itself. A typical example is the insertion of a cochlear implant which should ideally be done with minimum physical contact between the moving device and the cochlear canal walls or the basilar membrane. Because optical monitoring is not possible, alternative techniques for sub millimeter-scale distance control can be very useful for such procedures. The first requirement for distance control is distance sensing. We developed a novel approach to distance sensing based on the principles of scanning electrochemical microscopy (SECM). The SECM signal, i.e., the diffusion current to a microelectrode, is very sensitive to the distance between the probe surface and any electrically insulating object present in its proximity. With several amperometric microprobes fabricated on the surface of an insertable device, one can monitor the distances between different parts of the moving implant and the surrounding tissues. Unlike typical SECM experiments, in which a disk-shaped tip approaches a relatively smooth sample, complex geometries of the mobile device and its surroundings make distance sensing challenging. Additional issues include the possibility of electrode surface contamination in biological fluids and the requirement for a biologically compatible redox mediator.

A method based on a set of new theorems for the analysis of multichannel time series is described, based on precise Fourier transform and coherence analysis of the restored signals from a detailed set of frequency components. Magnetic field recordings of spontaneous and evoked activity by means of magnetic encephalography demonstrated that multichannel precise Fourier spectrum contains a very large set of harmonics with high coherence. The inverse problem can be solved with great precision based on coherent harmonics, so the technique is a promising platform of general analysis in brain imaging. The analysis method makes it possible to reconstruct sites and timing of electrical activity generated by both spontaneous and evoked brain function at different depths in the brain in the millisecond time range.

The present hardware circuit was designed as a fast and energy efficient motor control system based on cerebellar oscillatory neuron activity and network dynamics. Specifically, a hardware model of the olivo-cerebellar dynamics controlling vertebrate motor coordination is used to control movement in an underwater robotic vehicle. Single shot oscillatory phase resetting is used for instantaneous motor plant reorganization based on incoming sensory information. Such a rapid feedback mode, which is rapid enough to prevent animals from falling when they stumble, has been previously described in biological and mathematical papers (Pellionisz and Llinas, 1979, Velarde et al., 2002, 2004). In the present control system, the direction of the vehicle displacement is captured by a camera, and transformed into a phase shift modulation of sets of oscillatory elements that embody internal dynamics. This design provides a novel real time control platform for robotic control in three dimensions. (C) 2013 Elsevier B.V. All rights reserved. C1 [Porras, A.; Llinas, R.] NYU, Sch Med, Dept Neurosci & Physiol, New York, NY 10016 USA

Superfusion of the squid giant synapse with artificial seawater (ASW) based on isotonic saline containing oxygen nanobubbles (RNS60 ASW) generates an enhancement of synaptic transmission. This was determined by examining the postsynaptic response to single and repetitive presynaptic spike activation, spontaneous transmitter release, and presynaptic voltage clamp studies. In the presence of RNS60 ASW single presynaptic stimulation elicited larger postsynaptic potentials (PSP) and more robust recovery from high frequency stimulation than in control ASW. Analysis of postsynaptic noise revealed an increase in spontaneous transmitter release with modified noise kinetics in RNS60 ASW. Presynaptic voltage clamp demonstrated an increased EPSP, without an increase in presynaptic ICa(++) amplitude during RNS60 ASW superfusion. Synaptic release enhancement reached stable maxima within 5-10 min of RNS60 ASW superfusion and was maintained for the entire recording time, up to 1 h. Electronmicroscopic morphometry indicated a decrease in synaptic vesicle density and the number at active zones with an increase in the number of clathrin-coated vesicles (CCV) and large endosome-like vesicles near junctional sites. Block of mitochondrial ATP synthesis by presynaptic injection of oligomycin reduced spontaneous release and prevented the synaptic noise increase seen in RNS60 ASW. After ATP block the number of vesicles at the active zone and CCV was reduced, with an increase in large vesicles. The possibility that RNS60 ASW acts by increasing mitochondrial ATP synthesis was tested by direct determination of ATP levels in both presynaptic and postsynaptic structures. This was implemented using luciferin/luciferase photon emission, which demonstrated a marked increase in ATP synthesis following RNS60 administration. It is concluded that RNS60 positively modulates synaptic transmission by up-regulating ATP synthesis, thus leading to synaptic transmission enhancement.

The reflexological view of brain function (Sherrington, 1906) has played a crucial role in defining both the nature of connectivity and the role of the synaptic interactions among neuronal circuits. One implicit assumption of this view, however, has been that CNS function is fundamentally driven by sensory input. This view was questioned as early as the beginning of the last century when a possible role for intrinsic activity in CNS function was proposed by Thomas Graham Brow (Brown, 1911, 1914). However, little progress was made in addressing intrinsic neuronal properties in vertebrates until the discovery of calcium conductances in vertebrate central neurons leading dendritic electroresponsiveness (Llinas and Hess, 1976; Llinas and Sugimori, 1980a,b) and subthreshold neuronal oscillation in mammalian inferior olive (IO) neurons (Llinas and Yarom, 1981a,b). This happened in parallel with a similar set of findings concerning invertebrate neuronal system (Marder and Bucher, 2001). The generalization into a more global view of intrinsic rhythmicity, at forebrain level, occurred initially with the demonstration that the thalamus has similar oscillatory properties (Llinas and Jahnsen, 1982) and the ionic properties responsible for some oscillatory activity were, in fact, similar to those in the IO (Jahnsen and Llinas, 1984; Llinas, 1988). Thus, lending support to the view that not only motricity, but cognitive properties, are organized as coherent oscillatory states (Pare et al., 1992; Singer, 1993; Hardcastle, 1997; Llinas et al., 1998; Varela et al., 2001).

At perceptual threshold, some stimuli are available for conscious access whereas others are not. Such threshold inputs are useful tools for investigating the events that separate conscious awareness from unconscious stimulus processing. Here, viewing unmasked, threshold-duration images was combined with recording magnetoencephalography to quantify differences among perceptual states, ranging from no awareness to ambiguity to robust perception. A four-choice scale was used to assess awareness: "didn't see" (no awareness), "couldn't identify" (awareness without identification), "unsure" (awareness with low certainty identification), and "sure" (awareness with high certainty identification). Stimulus-evoked neuromagnetic signals were grouped according to behavioral response choices. Three main cortical responses were elicited. The earliest response, peaking at approximately 100 ms after stimulus presentation, showed no significant correlation with stimulus perception. A late response ( approximately 290 ms) showed moderate correlation with stimulus awareness but could not adequately differentiate conscious access from its absence. By contrast, an intermediate response peaking at approximately 240 ms was observed only for trials in which stimuli were consciously detected. That this signal was similar for all conditions in which awareness was reported is consistent with the hypothesis that conscious visual access is relatively sharply demarcated.

Synaptic plasticity in many regions of the central nervous system leads to the continuous adjustment of synaptic strength, which is essential for learning and memory. In this study, we show by visualizing synaptic vesicle release in mouse hippocampal synaptosomes that presynaptic mitochondria and, specifically, their capacities for ATP production are essential determinants of synaptic vesicle exocytosis and its magnitude. Total internal reflection microscopy of FM1-43 loaded hippocampal synaptosomes showed that inhibition of mitochondrial oxidative phosphorylation reduces evoked synaptic release. This reduction was accompanied by a substantial drop in synaptosomal ATP levels. However, cytosolic calcium influx was not affected. Structural characterization of stimulated hippocampal synaptosomes revealed that higher total presynaptic mitochondrial volumes were consistently associated with higher levels of exocytosis. Thus, synaptic vesicle release is linked to the presynaptic ability to regenerate ATP, which itself is a utility of mitochondrial density and activity.