Synaptic organization of retinotectal connections of the frog application of pulse triggered averaging

Pulse-triggered averaging technique was applied to retinotectal connections of the frog. An extracellular single unit was first isolated from the terminals of retinal fibers, and then intracellular responses were recorded from a tectal neuron in the vicinity of the extracellular recording electrode. Intracellular potentials in response to a moving stimulus were averaged by triggering with the isolated presynaptic impulses. The results show that "on-off" retinal fibers monosynaptically excite E-E type (EPSP at "on" and "off" of light) and EI-EI type (EPSP-IPSP at "on" and "off" of light). One of the E-E type neurons was identified as a large ganglionic neuron in layer 8.     

Restoration of the behavioral reactions of Rana esculenta frogs during regeneration of visual nerve fibers]

After sectioning the optic nerve, various forms of visually guided behaviour in the frog R. esculenta recover unevenly. Within 16--20 days, the onset of reactions was observed which are associated with perception of relatively large unmoving objects during active locomotion of animals (1); within 55 days -- avoidance reactions (2); within 90 days -- feeling reactions (3) were formed. In electrophysiological experiments it was shown that axons of the ganglionic retinal cells from various classes form contacts with the tectum also at different periods. The data obtained confirm the significance of specialized "canalization" of the retino--central nervous systems of transmission and evaluation of visual information for organization of separate forms of visually monitored behaviour in frogs.     

Microbial electrode sensor for alcohols

A microbial electrode consisting of immobilized microorganisms, a gas permeable Teflon membrane, and an oxygen electrode was prepared for the continuous determination of methyl and ethyl alcohols. Immobilized Trichosporon brassicae was employed for a microbial electrode sensor for ethyl alcohol. When a sample solution containing ethyl alcohol was injected into a microbial electrode system, the current of the electrode decreased markedly with time until a steady state was reached. The response time was within 10 min by the steady state method and within 6 min by the pulse method. A linear relationship was observed between the current decrease and the concentration of ethyl alcohol below 22.5 mg/liter. The current was reproducible within ± 6% of the relative error when a sample solution containing 16.5 mg/liter ethyl alcohol. The standard deviation was 0.5 mg/liter in 40 experiments. The selectivity of the microbial electrode sensor for ethyl alcohol was satisfactory. The microbial electrode sensor was applied to a fermentation broth of yeasts and satisfactory comparative results were obtained (correlation coefficient 0.98). The current output of the microbial electrode sensor was almost constant for more than three weeks and 2100 assays. A microbial electrode sensor using immobilized bacteria for methyl alcohol was also described.     

The androgen receptor mRNA is up-regulated by testosterone in both the harderian gland and thumb pad of the frog, Rana esculenta

³²P-labelled cDNA probe from plasmid containing rat androgen receptor (rAR) has been tested in hybridization experiments using RNAs from the Harderian gland and thumb pad of the edible frog, Rana esculenta. Northern blot analysis has shown a high degree of homology between the rAR cDNA and the frog androgen receptor mRNA (fAR mRNA); this has been supported by both the hybridization conditions (high stringency) and the molecular size of fAR mRNA which is quite similar to those described in mammals (9.4 kb). The role of androgens has been further investigated with respect to the kinetics of expression of fAR mRNA in in vivo experiments. In both the Harderian gland and thumb pad, testosterone has increased the levels of fAR mRNA as compared with the untreated groups. The use of cyproterone acetate (CPA) in combination with testosterone has resulted in a loss of the increase in fAR mRNA as compared to testosterone-treated groups, while CPA alone has resembled the control group. In primary cultures of frog Harderian gland and thumb pad cells, the steady-state levels of fAR mRNA have been increased in the cells exposed to testosterone as compared to those not exposed. These findings confirm that, in these androgen target tissues, testosterone exerts an up-regulation on its own receptors, increasing the accumulation of fAR mRNA in the same way as oestrogens up-regulate the expression of their own receptors in Xenopus liver and oviduct cells.     

Electrophysiological properties of isthmic neurons in frogs revealed by in vitro and in vivo studies

The frog nucleus isthmi (parabigeminal nucleus in mammals) is a visually responsive, cholinergic and anatomically well-defined group of neurons in the midbrain. It shares reciprocal topographic projections with the ipsilateral optic tectum (superior colliculus in mammals) and strongly influences visual processing. Anatomical and biochemical information indicates the existence of distinct neural populations within the frog nucleus isthmi, which raises the question: are there electrophysiological distinctions between neurons that are putatively classified by their anatomical and biochemical properties? To address this question, we measured frog nucleus isthmi neuron cellular properties in vitro and visual response properties in vivo. No evidence for distinct electrophysiological classes of neurons was found. We thus conclude that, despite the anatomical and biochemical differences, the cells of the frog nucleus isthmi respond homogeneously to both current injections and simple visual stimuli.     

A sputtered thin film of nanostructured Ni/Pt/Ti on Al2O3 substrate for ethanol sensing

A novel thin film ethanol sensor using sputtered Ni/Pt/Ti on an Al2O3 substrate as the working electrode in an alkaline solution was developed. Atomic force microscopy (AFM) and scanning electron microscopy (SEM) were used to characterize the nanostructure of nickel films. Sputtering deposition conditions for maximum catalytic efficiency, electrode selectivity, and reproducibility were discussed. The results showed that ethanol oxidation was more efficient on the sputtered Ni/Pt/Ti on an Al2O3 substrate electrode than that on the conventional nickel electrode. The optimal operating conditions to generate the sputtered Ni/Pt/Ti on the Al2O3 substrate electrode were: 45 min of Ni sputtering deposition time, and 50 W of Ni sputtering power. The results also indicated that the response time of the prepared ethanol sensor is 27 s and the best sensitivity is 3.08 microA microM(-1) cm(-2).     

Response characteristics of the BVP neuron model to periodic pulse inputs.

The characteristics of the BVP neuron model response to periodic pulse stimuli are investigated. Temporal patterns of the output of the model are analyzed as a function of the stimulus intensity and period. The BVP model exhibits the same chaotic behavior, and a Cantor function-like graph of the response frequency (mean firing rate) as in electrophysiological experiments. This shows that the BVP model describes the complicated response characteristics of the neuron at least qualitatively.     

Development of an ultra high sensitive tissue biosensor for determination of swellfish poisoning, tetrodotoxin

A simple tissue biosensor for measuring Na⁺ channel blockers such as tetrodotoxin (TTX) and saxitoxin (STX) has been developed. The membrane of frog bladder has Na⁺ channels which control the passage of Na⁺. It is well known that TTX blocks Na⁺ channels. The tissue biosensor consists of a Na⁺ electrode integrated within a flow cell. The tip of the electrode was covered with frog bladder membrane sandwiched between two sheets of cellulose acetate membrane, and the electrode was set in a flow cell.

A solution of 8% NaCl was carried in the cell and the output of the electrode allowed to stabilize. TTX was injected into the sensor system and measured from the inhibition ratio of the sensor peak output. One assay took approximately 5 min. The lower limit of detection was 86 fg. The continuous determination of TTX was feasible for 250 h in the presence of 0·003% NaN₃. A Linear correlation was obtained between TTX activities of F-niphobles and F-parudale determined by the methods of TTX sensor and mouse assay.     

The saltatory effect in vision.

Events in vision triggered by brief flashes in train prove to be strictly analogous to those generated in the skin by a succession of taps. A requisite condition for their appearance in vision is projection of the flashes into the peripheral retinal field. Experiments are described that establish the general relation between extent of saltatory leaping and degree of retinal eccentricity and between leaping and retinal subtense of stimulus patches. Additionally, a curious "dip" phenomenon and several qualitative experiments in which color has been used to "tag" the saltatory image are reported.     

ADPβS evokes microglia activation in the rabbit retina <Emphasis Type="Italic">in vivo</Emphasis>

To investigate whether stimulation of purinergic P2Y(1) receptors modulates the activation of microglial and Müller glial cells in the rabbit retina in vivo, adenosine 5'-O-(2-thiodiphosphate) (ADPbetaS; 2 mM in 100 mul saline), a non-hydrolyzable ADP analogue, was intravitreadly applied into control eyes or onto retinas that were experimentally detached from the pigment epithelium. Both retinal detachment and application of ADPssS onto control retinas induced phenotype alterations of the microglial cells (decrease of soma size, retraction of cell processes) and had no influence on microglial cell density. ADPssS application onto detached retinas accelerated the process retraction and resulted in a strongly decreased density of microglial cells. The effects of ADPssS on microglia density and phenotype in detached retinas were partially reversed by co-application of the selective inhibitor of P2Y(1) receptors, MRS-2317 (3 mM in 100 mul saline). ADPssS apparently did not influence Müller cell gliosis, as determined by electrophysiological and calcium imaging records. It is concluded that rabbit retinal microglial cells express functional P2Y(1) receptors in vivo, and that activation of these receptors stimulates phenotype alterations that are characteristical for microglia activation.     

Frame of reference transformations in motion perception during smooth pursuit eye movements

Smooth pursuit eye movements change the retinal image velocity of objects in the visual field. In order to change from a retinocentric frame of reference into a head-centric one, the visual system has to take the eye movements into account. Studies on motion perception during smooth pursuit eye movements have measured either perceived speed or perceived direction during smooth pursuit to investigate this frame of reference transformation, but never both at the same time. We devised a new velocity matching task, in which participants matched both perceived speed and direction during fixation to that during pursuit. In Experiment 1, the velocity matches were determined for a range of stimulus directions, with the head-centric stimulus speed kept constant. In Experiment 2, the retinal stimulus speed was kept approximately constant, with the same range of stimulus directions. In both experiments, the velocity matches for all directions were shifted against the pursuit direction, suggesting an incomplete transformation of the frame of reference. The degree of compensation was approximately constant across stimulus direction. We fitted the classical linear model, the model of Turano and Massof (2001) and that of Freeman (2001) to the velocity matches. The model of Turano and Massof fitted the velocity matches best, but the differences between de model fits were quite small. Evaluation of the models and comparison to a few alternatives suggests that further specification of the potential effect of retinal image characteristics on the eye movement signal is needed.     

Integration of videooculography and encephalography for investigation of visual selective attention in humans

An original method of EEG recording in combination with eye tracking was developed. Due to the precise synchronization (which can include any amount of various recording devices), it is now possible to record short-time electrophysiological patterns evoked by some visual stimulus. The key feature of the method is the possibility to study not only single effects of a stimulus but to extract electrophysiological records corresponding to some part of complex image scanning.     

体外循环血栓非侵入在线电阻抗成像方法

目的 在体外循环系统中,血栓的在线检测和可视化具有重要意义。本文提出了基于电阻抗成像（EIT）的体外循环血栓非侵入在线检测方法。方法 首先通过联合仿真研究了传感器尺寸对成像效果的影响。其次,根据仿真结果设计了直径为20 mm的16铜质电极EIT传感器,搭建了循环流动实验平台,并设计了静态和循环流动实验。使用尺寸为3～6 mm的猪血块代替血栓,将血块置于新鲜猪血样本中,采用Tikhonov正则化算法进行成像。将3 mm和5 mm的血块分别置于循环系统中,重建血块在传感器截面的大小和位置图像,并与高速相机拍摄结果进行对比。结果 仿真结果显示当目标物与传感器面积比（AR）不小于0.01时,传感器直径为20 mm和30 mm对应的图像相关系数（IC）均大于0.06,成像效果较好。静态成像结果显示,相对尺寸覆盖率误差（RCR）小于等于0.1。循环流动实验显示,血块经过传感器时,检测到归一化后的相对电导率变化值分别为80和200,结果显示该方法能够检测到循环系统中的血块。结论 该方法具有实时性和非侵入的优点,有望应用于体外血栓的检测。     

A neural computation for visual acuity in the presence of eye movements

Humans can distinguish visual stimuli that differ by features the size of only a few photoreceptors. This is possible despite the incessant image motion due to fixational eye movements, which can be many times larger than the features to be distinguished. To perform well, the brain must identify the retinal firing patterns induced by the stimulus while discounting similar patterns caused by spontaneous retinal activity. This is a challenge since the trajectory of the eye movements, and consequently, the stimulus position, are unknown. We derive a decision rule for using retinal spike trains to discriminate between two stimuli, given that their retinal image moves with an unknown random walk trajectory. This algorithm dynamically estimates the probability of the stimulus at different retinal locations, and uses this to modulate the influence of retinal spikes acquired later. Applied to a simple orientation-discrimination task, the algorithm performance is consistent with human acuity, whereas naive strategies that neglect eye movements perform much worse. We then show how a simple, biologically plausible neural network could implement this algorithm using a local, activity-dependent gain and lateral interactions approximately matched to the statistics of eye movements. Finally, we discuss evidence that such a network could be operating in the primary visual cortex.     

The retinal rod outer segment of the frog: detachment, isolation, phosphorus fractions and enzyme activity

Frog retinal rod outer segments were detached from dark adapted retinas by (1) agitation in frog Ringer's solution or (2) by crushing between two glass surfaces. The resulting suspensions were further purified by low and high speed centrifugation procedures in Ficoll density gradients. The density of the outer segments in Ficoll solutions was found to be 1.09. The large frog outer segments, unlike bovine outer segments, are not readily separated from nuclei, which were estimated to comprise 2.6–8% of the material, based on DNA analyses. The RNA/DNA ratio was 0.4–0.5, like that of neuronal nuclei. Representative enzymes of glycolysis (lactic dehydrogenase and glyceraldehyde phosphate dehydrogenase), phosphogluconate oxidation (glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase), the citric acid cycle (malic dehydrogenase) and ATP degradation (ATPase) were assayed. A major part of the malic dehydrogenase activity was probably due to inner segments attached to some of the outer segments. Glyceraldehyde phosphate dehydrogenase and glucose-6-phosphate dehydrogenase (but not lactic dehydrogenase) activities were lower in detached outer segments purified on Ficoll gradients than in samples of outer segment layer microdissected from freeze-dried sections of frog retina, as well as in whole retina. The data suggest that the activities of all the enzymes studied are intrinsically low in rod outer segments.     

Multivariate phenotypic selection on a complex sexual signal

Animal signals are complex, comprising multiple components that receivers may use to inform their decisions. Components may carry information of differing value to receivers, and selection on one component could modulate or reverse selection on another, necessitating a multivariate approach to estimating selection gradients. However, surprisingly few empirical studies have estimated the strength of phenotypic selection on complex signals with appropriate design and adequate power to detect nonlinear selection. We used phonotaxis assays to measure sexual selection on the advertisement signal of Cope's gray tree frog, Hyla chrysoscelis. Female preferences were assessed for five signal components using single‐ and two‐stimulus behavioral assays. Linear, quadratic, and correlational selection gradients were estimated from the single‐stimulus data. Significant directional selection is acting on call duration, call rate, pulse rate, and relative amplitude; stabilizing selection is acting on call duration and call rate. Under the two‐stimulus paradigm, conclusions were qualitatively different, revealing nonlinear selection on all components except call duration. For individual subjects, the outcomes of single‐ and two‐stimulus trials were frequently discordant, suggesting that the choice of testing paradigm may affect conclusions drawn from experiments.     

A Simple and Accurate Model to Predict Responses to Multi-electrode Stimulation in the Retina

Implantable electrode arrays are widely used in therapeutic stimulation of the nervous system (e.g. cochlear, retinal, and cortical implants). Currently, most neural prostheses use serial stimulation (i.e. one electrode at a time) despite this severely limiting the repertoire of stimuli that can be applied. Methods to reliably predict the outcome of multi-electrode stimulation have not been available. Here, we demonstrate that a linear-nonlinear model accurately predicts neural responses to arbitrary patterns of stimulation using in vitro recordings from single retinal ganglion cells (RGCs) stimulated with a subretinal multi-electrode array. In the model, the stimulus is projected onto a low-dimensional subspace and then undergoes a nonlinear transformation to produce an estimate of spiking probability. The low-dimensional subspace is estimated using principal components analysis, which gives the neuron’s electrical receptive field (ERF), i.e. the electrodes to which the neuron is most sensitive. Our model suggests that stimulation proportional to the ERF yields a higher efficacy given a fixed amount of power when compared to equal amplitude stimulation on up to three electrodes. We find that the model captures the responses of all the cells recorded in the study, suggesting that it will generalize to most cell types in the retina. The model is computationally efficient to evaluate and, therefore, appropriate for future real-time applications including stimulation strategies that make use of recorded neural activity to improve the stimulation strategy.     

Biosynthesis of Somatostatin-Like Immunoreactivity by Frog Retinas In Vitro

Abstract: Somatostatin-like immunoreactivity has been localized to a wide variety of central nervous system neurons, including the retina. We utilized the unique advantages the retina provides for in vitro studies of nerves to examine the biosynthesis of somatostatin. Extracts of frog retinas pulse-labeled with [³⁵S]cysteine for various time periods revealed uptake of radioactivity into material adsorbable by anti-somatostatin antibody linked to affinity beads. This uptake increased in a curvilinear fashion for 4 h and was inhibited by cycloheximide (0.2 m M ) or by boiling the retinas prior to labeling. Pulse-chase experiments revealed that affinity-adsorbable radioactivity from retinal extracts decreased with time of incubation in chase medium; 89% of this decrease could be accounted for by increases in the affinity-adsorbable radioactivity of the chase medium. Chromatography of the retinal extracts on Sephadex G50 (superfine) revealed four elution peaks, whereas only one peak, co-eluted with somatostatin-14, could be identified in the medium. Chromatographic elution patterns of affinity-adsorbable radioactivity from extracts of pulse-labeled retinas incubated in chase medium for various times showed a gradual shift of radioactivity from the earlier-eluting peaks to the later ones. These studies indicate that biosynthesis of somatostatin occurs in frog retinas in vitro. The retina may be a useful model for further study of peptidergic neurons.     

Asymmetrical dendritic fields of retinal ganglion cells

By use of Golgi chrome—silver impregnation, studies were made of the dendritic branchings of feline and frog ganglion cells. It was shown that besides the known varieties of ganglion cells there were asymmetrical neurones whose dendrites lay all to one side. Essential differences distinguished these ganglion cells in the cat from those in the frog, differences depending upon the architectonics of the inner plexiform layer, which is broad and subdivided into layers in the frog, and narrow in the cat. We discuss the possible role of neurones with a unilateral arrangement of dendrites in relation to know electrophysiological data on retinal detectors and the receptive fields of ganglion cells.Brain Institute, Academy of Medical Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 3, No. 3, pp. 301–307, May–June, 1971.     

Movement correction of digital sequence angiographies of the retina]

Retinal hemodynamics can be quantified from videoangiographic image sequences by digital image processing. Intensity changes of dye dilution curves provide dynamics parameters of the local retinal blood flow. The measuring points of dye dilution curves have to be fixed on identical image contents in each image of a complete image sequence. To obtain measurements for every pixel on the retinal surface a motion-compensated image sequence is required. A new method adapted to the compensation of eye motion and movement artifacts in Scanning Laser Ophthalmoscopy in long image sequences (300-500 images) is presented in this paper. To inhibit error propagation of time sequential motion estimation, the eye movement is divided into two dynamic movements components. The method presented permits compensation for eye motion in retinal fluorescein angiographic sequences. Owing to the short calculation times, this algorithm can be used in clinical routine.