Saumil S. Patel

Moving ahead through differential visual latency

The time it takes to transmit information along the human visual pathways introduces a substantial delay in the processing of images that fall on the retina. This visual latency might be expected to cause a moving object to be perceived at a position behind its actual one, disrupting the accuracy of visually guided motor actions such as catching or hitting, but this does not happen. It has been proposed that the perceived position of a moving object is extrapolated forwards in time to compensate for the delay in visual processing.

Principles of connectivity among morphologically defined cell types in adult neocortex

A census of neocortical neurons Despite the importance of the brains neocortex, we still do not completely understand the diversity and functional connections of its cell types. Jiang et al. recorded, labeled, and classified over 1200 interneurons and more than 400 pyramidal neurons in the mature mouse visual cortex. Fifteen major classes of interneurons fell into three types: some connect to all neurons, some connect to other interneurons, and some form synapses with pyramidal neurons. Science, this issue p. 10.1126/science.aac9462 The connections between more than 10,000 pairs of individually classified neurons in the visual cortex of adult mice are mapped. INTRODUCTION The intricate microcircuitry of the cerebral cortex is thought to be a critical substrate from which arise the impressive capabilities of the mammalian brain. Until now, our knowledge of the stereotypical connectivity in neocortical microcircuits has been pieced together from individual studies of the connectivity between small numbers of neuronal cell types. Here, we provide unbiased, large-scale profiling of neuronal cell types and connections to reveal the essential building blocks of the cortex and the principles governing their assembly into cortical circuits. Using advanced techniques for tissue slicing, multiple simultaneous whole-cell recording, and morphological reconstruction, we are able to provide a comprehensive view of the connectivity between diverse types of neurons, particularly among types of γ-aminobutyric acid–releasing (GABAergic) interneurons, in the adult animal. RATIONALE We took advantage of a method for preparing high-quality slices of adult tissue and combined this technique with octuple simultaneous, whole-cell recordings followed by an improved staining method that allowed detailed recovery of axonal and dendritic arbor morphology. These data allowed us to perform a census of morphologically and electrophysiologically defined neuronal types (primarily GABAergic interneurons) in neocortical layers 1, 2/3, and 5 (L1, L23, and L5, respectively) and to observe their connectivity patterns in adult animals. RESULTS Our large-scale, comprehensive profiling of neocortical neurons differentiated 15 major types of interneurons, in addition to two lamina-defined types of pyramidal neurons (L23 and L5). Cortical interneurons comprise two types in L1 (eNGC and SBC-like), seven in L23 (L23MC, L23NGC, BTC, BPC, DBC, L23BC, and ChC), and six in L5 (L5MC, L5NGC, L5BC, SC, HEC, and DC) (see the figure). Each type has stereotypical electrophysiological properties and morphological features and can be differentiated from all others by cell type–specific axonal geometry and axonal projection patterns. Importantly, each type of neuron has its own characteristic input-output connectivity profile, connecting with other constituent neuronal types with varying degrees of specificity in postsynaptic targets, laminar location, and synaptic characteristics. Despite specific connection patterns for each cell type, we found that a small number of simple connectivity motifs are repeated across layers and cell types defining a canonical cortical microcircuit. CONCLUSION Our comprehensive profiling of neuronal cell types and connections in adult neocortex provides the most complete wiring diagram of neocortical microcircuits to date. Compared with current genetic labels for cell class, which paint the cortex in broad strokes, our analysis of morphological and electrophysiological properties revealed new cell classes and allowed us to derive a small number of simple connectivity rules that were repeated across layers and cell types. This detailed blueprint of cortical wiring should aid efforts to identify specific circuit abnormalities in animal models of brain disease and may eventually provide a path toward the development of comprehensive circuit-based, cell type–specific interventions. Connectivity among morphologically defined cell types in adult neocortex. (A) Simultaneous octuple whole-cell recording to study connectivity followed by morphological reconstruction. (B) Synaptic connectivity between morphologically distinct types of neurons, including pyramidal (P) neurons

High-speed, random-access fluorescence microscopy: I. High-resolution optical recording with voltage-sensitive dyes and ion indicators

The design and implementation of a high-speed, random-access, laser-scanning fluorescence microscope configured to record fast physiological signals from small neuronal structures with high spatiotemporal resolution is presented. The laser-scanning capability of this nonimaging microscope is provided by two orthogonal acousto-optic deflectors under computer control. Each scanning point can be randomly accessed and has a positioning time of 3-5 microseconds. Sampling time is also computer-controlled and can be varied to maximize the signal-to-noise ratio. Acquisition rates up to 200k samples/s at 16-bit digitizing resolution are possible. The spatial resolution of this instrument is determined by the minimal spot size at the level of the preparation (i.e., 2-7 microns). Scanning points are selected interactively from a reference image collected with differential interference contrast optics and a video camera. Frame rates up to 5 kHz are easily attainable. Intrinsic variations in laser light intensity and scanning spot brightness are overcome by an on-line signal-processing scheme. Representative records obtained with this instrument by using voltage-sensitive dyes and calcium indicators demonstrate the ability to make fast, high-fidelity measurements of membrane potential and intracellular calcium at high spatial resolution (2 microns) without any temporal averaging.

Evidence of Cognitive Dysfunction after Soccer Playing with Ball Heading Using a Novel Tablet-Based Approach

Does frequent head-to-ball contact cause cognitive dysfunctions and brain injury to soccer players? An iPad-based experiment was designed to examine the impact of ball-heading among high school female soccer players. We examined both direct, stimulus-driven, or reflexive point responses (Pro-Point) as well as indirect, goal-driven, or voluntary point responses (Anti-Point), thought to require cognitive functions in the frontal lobe. The results show that soccer players were significantly slower than controls in the Anti-Point task but displayed no difference in Pro-Point latencies, indicating a disruption specific to voluntary responses. These findings suggest that even subconcussive blows in soccer can result in cognitive function changes that are consistent with mild traumatic brain injury of the frontal lobes. There is great clinical and practical potential of a tablet-based application for quick detection and monitoring of cognitive dysfunction.

Spatial and Temporal Properties of the Illusory Motion-Induced Position Shift for Drifting Stimuli

The perceived position of a stationary Gaussian window of a Gabor target shifts in the direction of motion of the Gabors carrier stimulus, implying the presence of interactions between the specialized visual areas that encode form, position, and motion. The purpose of this study was to examine the temporal and spatial properties of this illusory motion-induced position shift (MIPS). We measured the magnitude of the MIPS for a pair of horizontally separated (2 or 8deg) truncated-Gabor stimuli (carrier=1 or 4cpd sinusoidal grating, Gaussian envelope SD=18arc min, 50% contrast) or a pair of Gaussian-windowed random-texture patterns that drifted vertically in opposite directions. The magnitude of the MIPS was measured for drift speeds up to 16deg/s and for stimulus durations up to 453ms. The temporal properties of the MIPS depended on the drift speed. At low velocities, the magnitude of the MIPS increased monotonically with the stimulus duration. At higher velocities, the magnitude of the MIPS increased with duration initially, then decreased between approximately 45 and 75ms before rising to reach a steady-state value at longer durations. In general, the magnitude of the MIPS was larger when the truncated-Gabor or random-texture stimuli were more spatially separated, but was similar for the different types of carrier stimuli. Our results are consistent with a framework that suggests that perceived form is modulated dynamically during stimulus motion.

Neural network model of short-term horizontal disparity vergence dynamics

We present a neural network model of short-term dynamics of the human horizontal vergence system (HVS) and compare its predictions qualitatively and quantitatively with a large variety of horizontal disparity vergence data. The model consists of seven functional stages, namely: (1) computation of instantaneous disparity; (2) generation of a disparity map; (3) conversion of the disparity into a velocity signal; (4) push-pull integration of velocity to generate a position signal; (5) conversion of the position signal to motoneuron/plant activity for each eye; (6) gating of velocity overdrive signal to motoneuron/plant system; and finally (7) discharge path for position cells. Closed-loop (normal binocular viewing) symmetric step and staircase disparity vergence data were collected from three subjects and model parameters were determined to quantitatively match each subjects data. The simulated closed-loop as well as open-loop (disparity clamped viewing) symmetric step, sinusoidal, pulse, staircase, square and ramp wave responses closely resemble experimental results either recorded in our laboratory or reported in the literature. Where possible, the firing pattern of the neurons in the model have been compared to actual cellular recordings reported in the literature. The model provides insights into neural correlates underlying the dynamics of vergence eye movements. It also makes novel predictions about the human vergence system.

Dynamic Visual Stimulus Presentation in an Adaptive Optics Scanning Laser Ophthalmoscope

PURPOSE: To demonstrate the technology and application of synchronized laser modulation in the adaptive optics scanning laser ophthalmoscope (AOSLO), which makes it possible to deliver adaptive optics (AO) corrected stimuli to the retina of a living eye and to record the precise retinal location where the stimulus has landed. METHODS: The modification involves the development of custom software to control a high frequency pixel clock and a waveform generator board in synchrony with the scanning mirrors. The experiment involves a measurement of visual acuity with and without aberrations correction with AO. RESULTS: The system can project stimuli at a frame rate of 30 Hz with high sampling regotution (7.5 seconds of arc), thereby limiting the quality of the retinal image to the level of AO correction. Visual acuity in six subjects is improved on average by 33% after aberration correction across a 5.89-mm pupil. CONCLUSIONS: Dynamic visual stimulus presentation in an AOSLO works effectively and expands the scope of AOSLO applications.

Acute Hemodynamic Responses to Weightlessness During Parabolic Flight

Pilots and astronauts experience fluid shifts in variable gravity. Acute effects of fluid shifts on the cardiovascular system were monitored on NASAs KC‐135 aircraft during parabolic flight The variability of R‐R intervals in the electrocardiogram was measured as an indication of vagal cardiac neural activity. R‐R intervals were measured during the gravity transition from 2‐G to 0‐G produced by parabolic flight to assess the involvement of the autonomic nervous system in regulating the acute effects of fluid shifts. In seven subjects, a BoMed noninvasive continuous cardiac output monitor (NCCOM 3) monitored thoracic fluid index (TFI, ohms), heart rate (bpm), and cardiac output (1/min). Data were stored on a lap‐top computer with the subject in one of four postures: sitting, standing, supine, and semi‐supine, during one of four sets of eight to ten parabolas. Five seconds of data were averaged: before parabola onset (1.3‐G); parabola entry (1.9‐G); 0‐G; and parabola exit (1.7‐G). Three to eight parabolas were averaged for subjects in each posture; the mean for each posture was calculated. In each of five additional subjects, the coefficient of variation was calculated by dividing mean value by the standard deviation of 3 to 15 R‐R intervals. Eight to ten parabolas were averaged for each postural set. Compared with values collected before 0‐G, standing values during 0‐G showed that the thoracic fluid index decreased 2.5 ohms, heart rate decreased 22 bpm, and cardiac output increased 1 L/min. During sitting, thoracic fluid index decreased 1.25 ohms, heart rate decreased 10 bpm, whereas cardiac output increased 0.5 L/min. In the supine position, thoracic fluid index and heart rate were constant whereas cardiac output decreased 0.55 L/min. In the semi‐supine position, thoracic fluid index and heart rate were constant. Compared with values collected from 2‐G and 0‐G the coefficient of variation increased 66.4% in the standing position, 53.4% in the sitting position, and 43.3% in the semi‐supine position and decreased 11.6% in the supine position. The data indicated that cardiovascular changes are dependent on posture and gravity. During the four sets of parabolas in the four different postures, the greatest and smallest changes were observed in the standing and supine positions, respectively, during 0‐G. Fluid shifts from the legs to the thorax occurred during 0‐G in the supine and standing positions. The high values of the coefficient of variation at the onset of 0‐G suggest that vagal cardiac neural activity increases, but not significantly, in all positions except supine.

Autonomic effects on R-R variations of the heart rate in the squirrel monkey: an indicator of autonomic imbalance in conflict sickness.

To assess the contribution of the autonomic nervous system (ANS) to beat-to-beat (R-R interval) variations in the squirrel monkey, heart rate was analyzed after atropine, propranolol, carbachol, or isoproterenol administration, using a computer analyzing system. Heart rate variations were expressed as coefficient of variance (CV) of mean R-R interval. CV of R-R interval clearly was reduced by atropine but not by propranolol, indicating that the effect could be mediated through the parasympathetic nerve. CV of R-R interval was measured also under vestibulo-visual conflict (VVC) situation in pitch. Although CV of R-R interval increased during VVC, this variation almost was abolished after atropine administration. Monkeys with unilateral labyrinthectomy showed smaller variations than those without operation during exposure to VVC. CV of R-R interval is a useful parameter for the objective and quantitative evaluation of ANS function and may be a good indicator to demonstrate the severity of motion sickness without invasive methods.

Simultaneous optical recording of evoked and spontaneous transients of membrane potential and intracellular calcium concentration with high spatio-temporal resolution

We have developed a system for simultaneous optical recording of transients of membrane potential and intracellular calcium concentration from mammalian brain slice preparations with high spatio-temporal resolution. Simultaneous recording was achieved by using two dedicated photodetectors together with two fluorescent indicators. Specifically, the calcium-sensitive dye Calcium Orange and the voltage-sensitive dye RH-414 were selected because they have overlapping excitation spectra, but separable emission spectra. Transverse guinea pig hippocampal slices were double-loaded by bath application of the membrane-permeant form of Calcium Orange and RH-414. Transients of intracellular calcium concentration and membrane potential associated with evoked neural activity in hippocampal areas CA1 and CA3 were recorded. Furthermore, we have recorded calcium and voltage transients associated with spontaneous epileptiform activity induced by bath application of an epileptogenic drug, 4-aminopyridine. The use of photodiode matrices (10 x 10 elements each) as detectors gives the high spatial (200 x 200 microns/element with a 10 x objective) and temporal resolution (570 microseconds/frame). The recording system also includes a CCD camera for obtaining images of the preparation and overlaying the image with the optically detected signals. A software package has been developed for setting up the experimental protocol(s) and for collecting, processing, displaying, and analyzing the data in an user-friendly, windows-based environment.