Primary neuronal inhibitory responses of a chronically isolated slab of auditory cortex to intracortical stimulation in cats

Responses of 579 neurons in a slab of cerebral cortex (3 weeks after its isolation) to intracortical stimulation, with a distance of 0.5, 1.0, and 2.0 mm between recording and stimulating electrodes, were tested intercellularly and histological changes in a similar slab were studied in experiments on cats. Primary IPSPs were shown to develop in the chronically isolated slab in a much larger number of neurons than in the acutely isolated slab. Latent periods of IPSPs in all series of experiments did not exceed 10 msec, and most IPSPs were mono- and disynaptic. The amplitude and duration of the IPSPs were similar to those observed in the acutely isolated slab and intact auditory cortex in cats. It is concluded that local intracortical neuronal chains along which impulses evoking an inhibitory effect in the terminal neuron of the chain are transmitted are of relatively short length. Such chains may participate in local processing of incoming information. Analysis of the distribution of neurons responding by primary IPSPs by duration of their latent periods and depth in the slab in each series of experiments revealed a spatial and temporal mosaic of inhibitory responses in the chronically isolated slab of auditory cortex and showed that this mosaic is due to intracortical mechanisms.I. I. Mechnikov Odessa State University. Translated from Neirofiziologiya, Vol. 16, No. 2, pp. 152–161, March–April 1984.     

Prior knowledge about spatial pattern affects patch assessment rather than movement between patches in tactile-feeding mallard

1. Heterogeneity in food abundance allows a forager to concentrate foraging effort in patches that are rich in food. This might be problematic when food is cryptic, as the content of patches is unknown prior to foraging. In such case knowledge about the spatial pattern in the distribution of food might be beneficial as this enables a forager to estimate the content of surrounding patches. A forager can benefit from this pre-harvest information about the food distribution by regulating time in patches and/or movement between patches. 2. We conducted an experiment with mallard Anas platyrhynchos foraging in environments with random, regular, and clumped spatial configurations of full and empty patches. An assessment model was used to predict the time in patches for different spatial distributions, in which a mallard is predicted to remain in a patch until its potential intake rate drops to the average intake rate that can be achieved in the environment. A movement model was used to predict lengths of interpatch movements for different spatial distributions, in which a mallard is predicted to travel to the patch where it expects the highest intake rate. 3. Consistent with predictions, in the clumped distribution mallard spent less time in an empty patch when the previously visited neighbouring patch had been empty than when it had been full. This effect was not observed for the random distribution. This shows that mallard use pre-harvest information on spatial pattern to improve patch assessment. Patch assessment could not be evaluated for the regular distribution. 4. Movements that started in an empty patch were longer than movements that started in a full patch. Contrary to model predictions this effect was observed for all spatial distributions, rather than for the clumped distribution only. In this experiment mallard did not regulate movement in relation to pattern. 5. An explanation for the result that pre-harvest information on spatial pattern affected patch assessment rather than movement is that mallard move to the nearest patch where the expected intake rate is higher than the critical value, rather than to the patch where the highest intake rate is expected.     

Imaging cortical dynamics at high spatial and temporal resolution with novel blue voltage-sensitive dyes

Conventional imaging techniques have provided high-resolution imaging either in the spatial domain or in the temporal domain. Optical imaging utilizing voltage-sensitive dyes has long had the unrealized potential to achieve high resolution in both domains simultaneously, providing subcolumnar spatial detail with millisecond precision. Here, we present a series of developments in voltage-sensitive dyes and instrumentation that make functional imaging of cortical dynamics practical, in both anesthetized and awake behaving preparations, greatly facilitating exploration of the cortex. We illustrate this advance by analyzing the millisecond-by-millisecond emergence of orientation maps in cat visual cortex.     

Optimal movement between patches under incomplete information about the spatial distribution of food items

If the food distribution contains spatial pattern, the food density in a particular patch provides a forager with information about nearby patches. Foragers might use this information to exploit patchily distributed resources profitably. We model the decision on how far to move to the next patch in linear environments with different spatial patterns in the food distribution (clumped, random, and regular) for foragers that differ in their degree of information. An ignorant forager is uninformed and therefore always moves to the nearest patch (be it empty or filled). In contrast, a prescient forager is fully informed and only exploits filled patches, skipping all empty patches. A Bayesian assessor has prior knowledge about the content of patches (i.e. it knows the characteristics of the spatial pattern) and may skip neighbouring patches accordingly by moving to the patch where the highest gain rate is expected. In most clumped and regular distributions there is a benefit of assessment, i.e. Bayesian assessors achieve substantially higher long-term gain rates than ignorant foragers. However, this is not the case in distributions with less strong spatial pattern, despite the fact that there is a large potential benefit from a sophisticated movement rule (i.e. a large penalty of ignorance). Bayesian assessors do also not achieve substantially higher gain rates in environments that are relatively rich or poor in food. These results underline that an incompletely informed forager that is sensitive to spatial pattern should not always respond to existing pattern. Furthermore, we show that an assessing forager can enhance its long-term gain rate in highly clumped and some specific near-regular food distributions, by sampling the environment in slightly larger spatial units.     

Optimal movement between patches under incomplete information about the spatial distribution of food items

If the food distribution contains spatial pattern, the food density in a particular patch provides a forager with information about nearby patches. Foragers might use this information to exploit patchily distributed resources profitably. We model the decision on how far to move to the next patch in linear environments with different spatial patterns in the food distribution (clumped, random, and regular) for foragers that differ in their degree of information. An ignorant forager is uninformed and therefore always moves to the nearest patch (be it empty or filled). In contrast, a prescient forager is fully informed and only exploits filled patches, skipping all empty patches. A Bayesian assessor has prior knowledge about the content of patches (i.e. it knows the characteristics of the spatial pattern) and may skip neighbouring patches accordingly by moving to the patch where the highest gain rate is expected. In most clumped and regular distributions there is a benefit of assessment, i.e. Bayesian assessors achieve substantially higher long-term gain rates than ignorant foragers. However, this is not the case in distributions with less strong spatial pattern, despite the fact that there is a large potential benefit from a sophisticated movement rule (i.e. a large penalty of ignorance). Bayesian assessors do also not achieve substantially higher gain rates in environments that are relatively rich or poor in food. These results underline that an incompletely informed forager that is sensitive to spatial pattern should not always respond to existing pattern. Furthermore, we show that an assessing forager can enhance its long-term gain rate in highly clumped and some specific near-regular food distributions, by sampling the environment in slightly larger spatial units.     

Spike-timing precision and neuronal synchrony are enhanced by an interaction between synaptic inhibition and membrane oscillations in the amygdala

The basolateral complex of the amygdala (BLA) is a critical component of the neural circuit regulating fear learning. During fear learning and recall, the amygdala and other brain regions, including the hippocampus and prefrontal cortex, exhibit phase-locked oscillations in the high delta/low theta frequency band (～2-6 Hz) that have been shown to contribute to the learning process. Network oscillations are commonly generated by inhibitory synaptic input that coordinates action potentials in groups of neurons. In the rat BLA, principal neurons spontaneously receive synchronized, inhibitory input in the form of compound, rhythmic, inhibitory postsynaptic potentials (IPSPs), likely originating from burst-firing parvalbumin interneurons. Here we investigated the role of compound IPSPs in the rat and rhesus macaque BLA in regulating action potential synchrony and spike-timing precision. Furthermore, because principal neurons exhibit intrinsic oscillatory properties and resonance between 4 and 5 Hz, in the same frequency band observed during fear, we investigated whether compound IPSPs and intrinsic oscillations interact to promote rhythmic activity in the BLA at this frequency. Using whole-cell patch clamp in brain slices, we demonstrate that compound IPSPs, which occur spontaneously and are synchronized across principal neurons in both the rat and primate BLA, significantly improve spike-timing precision in BLA principal neurons for a window of ～300 ms following each IPSP. We also show that compound IPSPs coordinate the firing of pairs of BLA principal neurons, and significantly improve spike synchrony for a window of ～130 ms. Compound IPSPs enhance a 5 Hz calcium-dependent membrane potential oscillation (MPO) in these neurons, likely contributing to the improvement in spike-timing precision and synchronization of spiking. Activation of the cAMP-PKA signaling cascade enhanced the MPO, and inhibition of this cascade blocked the MPO. We discuss these results in the context of spike-timing dependent plasticity and modulation by neurotransmitters important for fear learning, such as dopamine.     

Odor-evoked responses in the olfactory center neurons in the terrestrial slug

The procerebrum (PC) of the terrestrial mollusk Limax is a highly developed second-order olfactory center consisting of two electrophysiologically distinct populations of neurons: nonbursting (NB) and bursting (B). NB neurons are by far the more numerous of the two cell types. They receive direct synaptic inputs from afferent fibers from the tentacle ganglion, the primary olfactory center, and also receive periodic inhibitory postsynaptic potentials (IPSPs) from B neurons. Odor-evoked activity in the NB neurons was examined using perforated patch recordings. Stimulation of the superior tentacle with odorants resulted in inhibitory responses in 45% of NB neurons, while 11% of NB neurons showed an excitatory response. The specific response was reproducible in each neuron to the same odorant, suggesting the possibility that activity of NB neurons may encode odor identity. Analysis of the cycle-averaged membrane potential of NB neurons revealed a correlation between the firing rate and the membrane potential at the plateau phase between IPSPs. Also, the firing rate of NB neurons was affected by the frequency of the IPSPs. These results indicate the existence of two distinct mechanisms for the regulation of NB neuron activity.     

High-definition mapping of neural activity using voltage-sensitive dyes

The distribution of patterns of activity in different brain structures has been related to the encoding and processing of sensory information. Consequently, it is important to be able to image the distribution of these patterns to understand basic brain functions. The spatial resolution of voltage-sensitive dye (VSD) methods has recently been enhanced considerably by the use of video imaging techniques. The main factor that now hampers the resolution of VSD patterns is the inherent limitation of the optical systems. Unfortunately, the intrinsic characteristics of VSD images impose important limitations that restrict the use of general deconvolution techniques. To overcomes this problem, in this study an image restoration procedure has been implemented that takes into consideration the limiting characteristics of VSD signals. This technique is based on applying a set of imaging processing steps. First, the signal-to-noise (S/N) ratio of the images was improved to avoid an increase in the noise levels during the deconvolution procedures. For this purpose, a new filter technique was implemented that yielded better results than other methods currently used in optical imaging. Second, focal plane images were deconvolved using a modification of the well-known nearest-neighbor deconvolution algorithm. But to reduce the light exposure of the preparation and simplify image acquisition procedures, adjacent image planes were modeled according to the in-focus image planes and the empirical point spread function (PSF) profiles. Third, resulting focal plane responses were processed to reduce the contribution of optical responses that originate in distant image planes. This method was found to be satisfactory under simulated and real experimental conditions. By comparing the restored and unprocessed images, it was clearly demonstrated that this method can effectively remove the out-of-focus artifacts and produce focal plane images of better quality. Evaluations of the tissue optical properties allowed assessment of the maximum practical optical section thickness using this deconvolution technique in the optical system tested. Determination of the three-dimensional PSF permitted the correct application of deconvolution algorithms and the removal of the contaminating light arising from adjacent as well as distant optical planes. The implementation of this deconvolution approach in salamander olfactory bulb allowed the detailed study of the laminar distribution of voltage-sensitive changes across the bulb layer. It is concluded that (1) this deconvolution procedure is well suited to deconvolved low-contrast images and offers important advantages over other alternatives; (2) this method can be properly used only when the tissue optical properties are first determined; (3) high levels of light scattering in the tissue reduce the optical section capabilities of this technique as well as other deconvolution procedures; and (4) use of the highest numerical aperture in the objectives is advisable because this improves not only the light-collecting efficiency to detect poor-contrast images, but also the spatial frequency differences between adjacent image planes. Under this condition it is possible to overcome some of the limitations imposed by the light scattering/birefringence of the tissue.     

VSDI: a new era in functional imaging of cortical dynamics

During the last few decades, neuroscientists have benefited from the emergence of many powerful functional imaging techniques that cover broad spatial and temporal scales. We can now image single molecules controlling cell differentiation, growth and death; single cells and their neurites processing electrical inputs and sending outputs; neuronal circuits performing neural computations in vitro; and the intact brain. At present, imaging based on voltage-sensitive dyes (VSDI) offers the highest spatial and temporal resolution for imaging neocortical functions in the living brain, and has paved the way for a new era in the functional imaging of cortical dynamics. It has facilitated the exploration of fundamental mechanisms that underlie neocortical development, function and plasticity at the fundamental level of the cortical column.     

Optical recordings of membrane potential using genetically targeted voltage-sensitive fluorescent proteins

Optical imaging of electrical activity using voltage-sensitive dyes has been envisaged for many years as a powerful method to investigate multineuronal representation of information processing in brain tissue. This article describes the advent of novel genetically targeted voltage-sensitive fluorescent proteins. This new class of membrane voltage sensors overcomes previous limitations related to the nonselective staining of membranes associated with conventional voltage-sensitive dyes. Here, we discuss the methodology, applications, and potential advantages of this novel technique.     

Odor‐evoked responses in the olfactory center neurons in the terrestrial slug

The procerebrum (PC) of the terrestrial mollusk Limax is a highly developed second‐order olfactory center consisting of two electrophysiologically distinct populations of neurons: nonbursting (NB) and bursting (B). NB neurons are by far the more numerous of the two cell types. They receive direct synaptic inputs from afferent fibers from the tentacle ganglion, the primary olfactory center, and also receive periodic inhibitory postsynaptic potentials (IPSPs) from B neurons. Odor‐evoked activity in the NB neurons was examined using perforated patch recordings. Stimulation of the superior tentacle with odorants resulted in inhibitory responses in 45% of NB neurons, while 11% of NB neurons showed an excitatory response. The specific response was reproducible in each neuron to the same odorant, suggesting the possibility that activity of NB neurons may encode odor identity. Analysis of the cycle‐averaged membrane potential of NB neurons revealed a correlation between the firing rate and the membrane potential at the plateau phase between IPSPs. Also, the firing rate of NB neurons was affected by the frequency of the IPSPs. These results indicate the existence of two distinct mechanisms for the regulation of NB neuron activity. © 2003 Wiley Periodicals, Inc. J Neurobiol 58: 369–378, 2004     

Fluorescence imaging of local membrane electric fields during the excitation of single neurons in culture.

The spatial distribution of depolarized patches of membrane during the excitation of single neurons in culture has been recorded with a high spatial resolution (1 micron2/pixel) imaging system based on a liquid-nitrogen-cooled astronomical camera mounted on an inverted microscope. Images were captured from rat nodose neurons stained with the voltage-sensitive dye RH237. Conventional intracellular microelectrode recordings were made in synchrony with the images. During an action potential the fluorescence changes occurred in localized, unevenly distributed membrane areas, which formed clusters of depolarized sites of different sizes and intensities. When fast conductances were blocked by the addition of tetrodotoxin, a reduction in the number and the intensities of the depolarized sites was observed. The blockade by tetrodotoxin of voltage-clamped neurons also reduced the number of depolarized sites, although the same depolarizing voltage step was applied. Similarly, when a voltage-clamped neuron was depolarized by a constant-amplitude voltage step, the number of depolarized sites varied according to the degree of activation of the voltage-sensitive channels, which was modified by changing the holding potential. These results suggest that the spatial patterns of depolarization observed during excitation are related to the operations of ionic channels in the membrane.     

Confocal ratiometric voltage imaging of cultured human keratinocytes reveals layer-specific responses to ATP

Recent evidencesuggests that changes in membrane potential influence the proliferationand differentiation of keratinocytes. To further elucidate the role ofchanges in membrane potential for their biological fate, the electricalbehavior of keratinocytes needs to be studied under complex conditionssuch as multilayered cultures. However, electrophysiological recordingsfrom cells in the various layers of a complex culture would beextremely difficult. Given the high spatial resolution of confocalimaging and the availability of novel voltage-sensitive dyes, wecombined these methods in an attempt to develop a viable alternativefor recording membrane potentials in more complex tissue systems. As afirst step, we used confocal ratiometric imaging of fluorescence resonance energy transfer (FRET)-based voltage-sensitive dyes. We thenvalidated this approach by comparing the optically recorded voltagesignals in HaCaT keratinocytes with the electrophysiological signalsobtained by whole cell recordings of the same preparation. Wedemonstrate 1) that optical recordings allow precisemultisite measurements of voltage changes evoked by the extracellularsignaling molecules ATP and bradykinin and 2) thatresponsiveness to ATP differs in various layers of cultured keratinocytes.

     

Reducing aberration by inverse filtering in Fresnel zone plate transducers

Transducers employing a spatial electrode pattern in the form of a Fresnel zone plate (FZP) are useful in acoustic imaging systems for focusing an ultrasonic beam. A technique has been developed to predict the spatial acoustic output of such a transducer from a knowledge of the spatial electrode pattern. It has been shown that, due to mechanical aberration, sharp edges in the electrode pattern are not reproduced in the output pattern. The mechanism producing the aberration can be modeled as a low-pass spatial filter. A desired acoustic spatial pattern may be generated by a synthesized electrode pattern obtained through inverse filtering. The approach is to assume that the desired acoustic pattern is the output of a linear spatial system and then to synthesize the electrode pattern that would produce it by multiplying it with the inverse of the aberrating low-pass filter. The synthesized electrode pattern will then generate an acoustic pattern which replicates the desired acoustic pattern very closely.     

Spatial and temporal predictions of moose winter distribution

Herbivores are usually distributed unevenly across the landscape often because of variation in resource availability. We used zero-inflated generalised additive models (to account for data with a high number of zeros) that include georeferences to predict winter distribution of a large herbivore (moose Alces alces). Moose distribution was analysed in relation to forage availability and distance to neighbouring sites. Our results showed that the ability to explain moose distribution indexed by pellet count data at a local scale increased when spatial information (longitude and latitude) was added to the model compared to the model only including food availability. By using the relationship between moose and forage distribution, and the spatial information, we predicted patch choice by moose reasonably well in 2 out of 4?years. However, the distribution of moose was also influenced by weather conditions, as it was most clumped in the year with most snow. In conclusion, our study lends support for a non-linear approach using georeferences for a comprehensive understanding of herbivore distribution at a small scale. This result also indicates that the use of a certain patch by moose not only depends on the selected patch itself but is also influenced by the neighbouring patch and factors at a larger spatial scale, such as moose management influencing the density above moose home range level. The relatively high proportion of unexplained variation suggests that the use of a certain patch is also influenced by other factors such as topography, predation, competition, weather conditions, and wildlife management strategies.     

Sex‐related spatial patterns of Poa ligularis in relation to shrub patch occurrence in northern Patagonia

Abstract. Poa ligularis is a dioecious species and a valuable forage plant which is widespread in the arid steppe of northern Patagonia (Argentina). The vegetation in these areas consists of a system of perennial plant patches alternating with bare soil areas defining contrasting micro‐environments. We hypothesized that (1) male and female individuals of P. ligularis are spatially segregated in different micro‐environments, (2) the intensity of spatial segregation of sexes depends on plant structure and (3) spatial segregation of sexes is enhanced by competitive interactions between the sexes within the vegetation patches. We analysed the spatial distribution of female and male individuals in relation to the spatial pattern of vegetation in two areas differing in their vegetation structure. The location of P. ligularis within patches where either male, female or both sexes occurred was also analysed. The results indicate that different patterns of spatial distribution of sexes of P. ligularis may be found at the community level depending on the dominant life forms and geometric structure of plant patches. Where patches are of a lower height, with a high internal patch cover, individuals of both sexes are concentrated within patch canopies. In sites characterized by large, tall patches and less internal patch cover suitable microsites for female and male P. ligularis occur both within and outside the patch with males located at further distances from the patch edge. Where the patch is large and tall enough to allow the establishment of males and females at relatively high numbers, males occupy the patch periphery or even colonize the interpatch bare soil. These spatial patterns are consistent with selective traits in which females better tolerate intraspecific competition than males, while males tolerate wider fluctuations in the physical environment (soil moisture, nitrogen availability, wind intensity, etc.).     

Chemical mapping of mammalian cells by atom probe tomography

In atom probe tomography (APT), a technique that has been used to determine 3D maps of ion compositions of metals and semiconductors at sub-nanometer resolutions, controlled emissions of ions can be induced from needle-shaped specimens in the vicinity of a strong electric field. Detection of these ions in the plane of a position sensitive detector provides two-dimensional compositional information while the sequence of ion arrival at the detector provides information in the third dimension. Here we explore the use of APT technology for imaging biological specimens. We demonstrate that it is possible to obtain 3D spatial distributions of cellular ions and metabolites from unstained, freeze-dried mammalian cells. Multiple peaks were reliably obtained in the mass spectrum from tips with diameters of ~50 nm and heights of ~200 nm, with mass-to-charge ratios (m/z) ranging from 1 to 80. Peaks at m/z 12, 23, 28 and 39, corresponding to carbon, sodium, carbonyl and potassium ions respectively, showed distinct patterns of spatial distribution within the cell. Our studies establish that APT could become a powerful tool for mapping the sub-cellular distribution of atomic species, such as labeled metabolites, at 3D spatial resolutions as high as ~1 nm.     

Aggregations from using inadvertent social information: a form of ideal habitat selection

Social information in breeding site selection has received extensive study; however, few attempts have been made to link this process to pre‐existing models. We examine the importance of social information to three pertinent models of habitat selection that describe breeding aggregations and spatial patterns: 1) the ideal despotic distribution (IDD) which considers conspecific competition and habitat availability, 2) the perceptual constraints model which accounts for patch selection when animals experience a threshold of undetectable difference in quality, and 3) the “neighbourhood model” which predicts concordance between resources and settlers can be disrupted by conspecific attraction when resources are patchy. These models all predict initial settlers will select a high quality patch first. However, their predictions of subsequent settlement behaviour in remaining patches differ: the IDD predicts subsequent settlers will be distributed regularly, the perceptual constraints model predicts a random distribution, and the neighbourhood model predicts clustering from conspecific attraction. We examined which model best described settlement patterns of bobolink Dolichonyx oryzivorus and savannah sparrow Passerculus sandwichensis, in the context of social information. We observed settlement timing, quantified available resources, and determined where they occurred in the highest (local population “core”) and lowest densities (local population “periphery”). We then assessed whether individuals in the periphery settled in greater concordance with resources or conspecific presence. Core territories were clustered strongly on relevant resources, and these territory holders were older than in the periphery. Peripheral territories were likewise clustered but did not always co‐occur with the best available resources, matching the neighbourhood model prediction that social information may not always direct them to the best sites available. This suggests older individuals used their own experience to locate ideal habitat, whereas younger individuals attempted to aggregate on seemingly ideal habitat by using conspecific location; such information asymmetry due to age can be viewed as an “ideal aggregative distribution”.     

Spatio-temporal dynamics of odor representations in the mammalian olfactory bulb

We explored the spatio-temporal dynamics of odor-evoked activity in the rat and mouse main olfactory bulb (MOB) using voltage-sensitive dye imaging (VSDI) with a new probe. The high temporal resolution of VSDI revealed odor-specific sequences of glomerular activation. Increasing odor concentrations reduced response latencies, increased response amplitudes, and recruited new glomerular units. However, the sequence of glomerular activation was maintained. Furthermore, we found distributed MOB activity locked to the nasal respiration cycle. The spatial distribution of its amplitude and phase was heterogeneous and changed by sensory input in an odor-specific manner. Our data show that in the mammalian olfactory bulb, odor identity and concentration are represented by spatio-temporal patterns, rather than spatial patterns alone.     

Optical single-channel recording: imaging Ca2+ flux through individual N-type voltage-gated channels expressed in Xenopus oocytes

Functional studies of single membrane ion channels were made possible by the introduction of the patch-clamp technique, which allows single-channel currents to be measured with unprecedented resolution. Nevertheless, patch clamping has some limitations: including the need for physical access of the patch pipette, possible disruption of local cellular architecture, inability to monitor multiple channels, and lack of spatial information. Here, we demonstrate the use of confocal fluorescence microscopy as a non-invasive technique to optically monitor the gating of individual Ca2+ channels. Near-membrane fluorescence signals track the gating of N-type Ca2+ channels with a kinetic resolution of about 10ms, provide a simultaneous and independent readout from several channels, and allow their locations to be mapped with sub-micrometer spatial resolution. Optical single-channel recording should be applicable to diverse voltage- and ligand-gated Ca2+-permeable channels, and has the potential for high-throughput functional analysis of single channels.