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Biomedical subjects

K Kirschfeld

Publications and source records attributed to K Kirschfeld.

At least 19 recordsLinked to original sources

Effects of volatile anaesthetics on the membrane potential and ion channels of cultured neocortical astrocytes.

Volatile anaesthetics cause changes in the membrane resting potential of central neurons. This effect probably arises from actions on neuronal ion channels, but may also involve alterations in the ion composition of the extracellular space. Since glial cells play a key role in regulating the extracellular ion composition in the brains of mammals, we analyzed the effects of halothane, isoflurane and enflurane on the membrane conductances and ion channels of cultured cortical astrocytes. Astrocytes were dissociated from the neocortex of 0-2-day old rats and grown in culture for 3-4 weeks. Anaesthetic-induced changes in the membrane potential were recorded in the whole cell current-clamp configuration of the patch-clamp technique. We further studied the effects of halothane and enflurane on single ion channels in excised membrane patches. At concentrations corresponding to 1-2 MAC (1 MAC induces general anaesthesia in 50% of the patients and rats), membrane potentials recorded in the presence of enflurane, isoflurane and halothane did not differ significantly from the control values. At higher concentrations, effects of enflurane and halothane, but not of isoflurane, were statistically significant. Single-channel recordings revealed that halothane and enflurane activated a high conductance anion channel, which possibly mediated the effects observed during whole cell recordings. In less than 10% of the membrane patches, volatile anaesthetics either increased or decreased the mean open time of K+-selective ion channels without altering single-channel conductances. In summary, it seems unlikely that the actions of volatile anaesthetics described here are involved in the state of general anaesthesia. Statistically significant effects occurred at concentrations ten times higher than those required to cause half-maximal depression of action potential firing of neocortical neurons in cultured brain slices. However, it cannot be excluded that the changes observed in the membrane conductance of cortical astrocytes disturb the physiological function of these cells, thereby influencing the membrane resting potential of neurons.

Anesthetics, Inhalation

Course control and tracking: orientation through image stabilization.

Course control and tracking are based on visual detection of the position and movement of objects. A disadvantage of biological movement detectors is that they cannot provide a signal proportional to the speed at which the image of an object moves over the retina. Other image parameters, such as brightness, contrast, and texture, strongly affect the magnitude of the detectors' output signals. To function well, the optomotor control circuit must solve these problems. One possible solution, realized in Diptera, is the principle of "gain control by feedback oscillations" described in this chapter. The optomotor system serves for course control by stabilizing the image of the visual panorama on the eye, and for tracking a moving object by stabilizing the object's image on the eye. When an object moves in front of a structured background, it is impossible for the images of both object and background to be stabilized simultaneously. Arthropods and vertebrates usually employ the same strategy to cope with this problem: saccadic tracking. In Diptera, the neural substrate for saccadic tracking is partially understood.

Animals

Effects of volatile anaesthetics on spontaneous action potential firing of cerebellar Purkinje cells in vitro do not follow the Meyer-Overton rule.

We have investigated in rat brain slices the effects of the volatile anaesthetics enflurane, isoflurane and halothane on spontaneous discharge patterns and mean firing rates of cerebellar Purkinje cells. In the absence of these anaesthetics, Purkinje cells fired bursts of action potentials separated by quiescent periods lasting less than 2 s. Mean discharge rates were 10.8 (SEM 0.4) Hz at 23 +/- 1 degrees C and 25.6 (1.2) Hz at 35 +/- 1 degrees C. The agents exhibited qualitatively different effects when applied at concentrations corresponding to 1-3 MAC. Enflurane markedly lengthened burst and inter-burst durations. Isoflurane acted in a similar manner, but effects were less pronounced. In contrast with isoflurane and enflurane, halothane shortened burst durations. At concentrations corresponding to 1-1.5 MAC, halothane, isoflurane and enflurane significantly depressed action potential firing by 15-30% (P < 0.05). Enflurane 1.2 mmol litre-1 (2.0 MAC), isoflurane 0.9 mmol litre-1 (2.8 MAC) and halothane 0.9 mmol litre-1 (3.8 MAC) depressed spontaneous spike rates by 50%. The changes in discharge patterns and the concentration-dependent decrease in the firing rates were similar at 23 +/- 1 degrees C and 35 +/- 1 degrees C. In summary, we observed that neither the anaesthetic-induced alterations in spontaneous discharge patterns nor the EC50 values of the concentration-dependent depression of the mean firing rates were in accordance with the Meyer-Overton rule. However, at clinically relevant concentrations, depression of average spike rates did not differ significantly between the anaesthetics and thus followed the rule. Our results suggest that anaesthetic actions, which are in accordance with the rule, are frequently masked by several side effects.

Action Potentials

Cortical oscillations and the origin of express saccades.

The latencies of visually guided saccadic eye movement can form bimodal distributions. The 'express saccades' associated with the first mode of the distributions are thought to be generated via an anatomical pathway different from that for the second mode, which comprises regular saccades. The following previously published observations are the basis for a new alternative model of these effects: (i) visual stimuli can cause oscillations to appear in the electroencephalogram; (ii) visual stimuli can cause a negative shift in the electroencephalogram that lasts for several hundreds of milliseconds; and (iii) negativity in the electroencephalogram can be associated with reduced thresholds of cortical neurons to stimuli. In the new model both express and regular saccades are generated by the same anatomical structures. The differences in saccadic latency are produced by an oscillatory reduction of a threshold in the saccade-generating pathway that is transiently produced under certain stimulus paradigms. The model has implications regarding the functional significance of spontaneous and stimulus-induced oscillations in the central nervous system.

Behavior

Visually elicited head rotation in pigeons (Columba livia).

Horizontal rotational head movements were video-taped from pigeons standing freely in a rotating cylinder. The cylinder carried vertically striped patterns approximating a sinusoidally modulated horizontal intensity distribution. We altered systematically various stimulus parameters: spatial wavelength and contrast of the pattern, angular velocity of the pattern motion and mode of motion onset. We found: (1) both gradual acceleration of the patterned cylinder as well as immediate onset of pattern motion elicit the sequence of smooth following and saccadic resetting movement typical of the rotational "stare" head nystagmus; (2) in experiments with rapid onset of pattern motion, velocity of the smooth following response gradually increases to its steady-state level over a period of about 10 sec; (3) the saccadic head rotations are not stereotyped: larger and shorter saccades follow in an irregular sequence, saccadic velocity and average size varies with stimulus conditions; (4) in the range of 0.9-95 deg/sec, the velocity of the following phase increases in parallel with stimulus speed; (5) in the range of spatial wavelengths of the striped patterns from 6 to 45 deg, at a given drum velocity, patterns of short wavelengths elicit optokinetic head rotations with higher gain (head velocity/drum velocity) than patterns of long wavelengths; (6) response velocity increases with pattern contrast (Michaelson contrast 5, 32 and 75%), following approximately a logarithmic relation; (7) our results on rotational optokinetic head movements support the notion that the neural mechanism underlying motion detection operates like a correlation mechanism.

Animals

Tracking of small objects in front of a textured background by insects and vertebrates: phenomena and neuronal basis.

To follow visually a small object moving in front of a textured background, insects and vertebrates can employ a similar strategy: saccadic tracking. In the case of vertebrates, the neural components that generate this behavior are not known in detail. The neural substrate of optomotor behavior in Diptera is relatively well understood. Here a model developed from the dipteran data is found to be capable of saccadic tracking. It is characterized by the following components and functions: (1) Two subsystems contribute to the response, a small-field tracking system and a large-field compensatory optomotor system, as suggested previously (Egelhaaf et al. 1988). (2) Both systems need to be suppressed during saccadic rotation. In the small-field system, the suppression, close to the visual input, is mediated by the activity of the large-field system. In the large-field system, suppression, close to the motor output, is due to efferent signals from the saccade generator. A similar model could also apply to vertebrates. Two implications of the present model are that saccadic tracking does not require object identification, and under saccadic tracking it is the background rather than the object that is stabilized on the retina. If objects are identified under these conditions, this must occur even though their image is not stabilized on the retina.

Animals

Genetic dissection of light-induced Ca2+ influx into Drosophila photoreceptors.

Invertebrate photoreceptors use the inositol-lipid signaling cascade for phototransduction. A useful approach to dissect this pathway and its regulation has been provided by the isolation of Drosophila visual mutants. We measured extracellular changes of Ca2+ [delta Ca2+]o in Drosophila retina using Ca(2+)-selective microelectrodes in both the transient receptor potential (trp) mutant, in which the calcium permeability of the light-sensitive channels is greatly diminished and in the inactivation-but-no-afterpotential C (inaC) mutant which lacks photoreceptor-specific protein kinase C (PKC). Illumination induced a decrease in extracellular [Ca2+] with kinetics and magnitude that changed with light intensity. Compared to wild-type, the light-induced decrease in [Ca2+]o (the Ca2+ signal) was diminished in trp but significantly enhanced in inaC. The enhanced Ca2+ signal was diminished in the double mutant inaC;trp indicating that the effect of the trp mutation overrides the enhancement observed in the absence of eye-PKC. We suggest that the decrease in [Ca2+]o reflects light-induced Ca2+ influx into the photoreceptors and that the trp mutation blocks a large fraction of this Ca2+ influx, while the absence of eye specific PKC leads to enhancement of light-induced Ca2+ influx. This suggestion was supported by Ca2+ measurements in isolated ommatidia loaded with the fluorescent Ca2+ indicator, Ca Green-5N, which indicated an approximately threefold larger light-induced increase in cellular Ca2+ in inaC relative to WT. Our observations are consistent with the hypothesis that TRP is a light activated Ca2+ channel and that the increased Ca2+ influx observed in the absence of PKC is mediated mainly via the TRP channel.

Animals

Motion sensitivity in the nucleus of the basal optic root of the pigeon.

1. Single-unit responses to large-field movement (angular velocity, w = 0.25-42 degrees/s) of sine-wave gratings of different spatial wavelength (lambda = 5.2-41 degrees) and contrast have been recorded in the nucleus of the basal optic root (nBOR) of the accessory optic system (AOS) of the pigeon. 2. The steady-state response to moving sine-wave gratings increases with increasing contrast to reach a saturation level at 25%. 3. Generally the steady-state responses of the cells passed through a maximum when stimulated at various velocities. In 12 of the 15 cells tested with six different velocities and four different spatial wavelengths, the location of the response maximum on the velocity scale depended on the spatial wavelength (lambda) used. That is, in these cells the response depends on the temporal frequency (tf = w/lambda) of the stimulus and not on its velocity alone. This is in agreement with the prediction of the theory of motion detection according to the basic version of the correlation scheme. 4. The temporal frequency for maximal response of individual cells shifts to higher values when the contrast of the sine-wave gratings is reduced to 5%. 5. The steady-state response of 16 of the recorded directional selective cells (53) is modulated with the temporal frequency of the stimulus, regardless of the phase of the grating at the beginning of its movement. 6. In phasic-tonically responding cells, the phasic response peak decays to the steady-state level with a time constant that becomes shorter as the temporal frequency of the stimulus increases. 7. The basic version of the correlation scheme includes only the time constant of one low-pass filter. Therefore the phasic response is expected to decay to the steady-state level with one and the same time constant, and the position of the maximal response on the temporal frequency scale should not be influenced by a change of pattern contrast. According to the model, phase-dependent modulations of the steady-state response should occur only when the spatial wavelength of the stimulus pattern is large compared with the sampling base of the underlying detector. Consequently the results given in points 4-6 cannot be described by a basic version of the correlation scheme.

Acceleration

Spectral tuning of rhodopsin and metarhodopsin in vivo.

Color vision is dependent upon the expression of spectrally distinct forms of rhodopsin in different photoreceptor cells. To identify the structural features of rhodopsin that regulate spectral sensitivity and absorption in vivo, we have constructed a series of chimeric Drosophila rhodopsin molecules, derived from a blue- and a violet-sensitive rhodopsin, and used P element-mediated germline transformation to generate transgenic flies that express the modified pigments in the R1-R6 photoreceptor cells of the compound eye. Our analysis of these animals indicates that multiple regions of the opsin protein are involved in regulating rhodopsin spectral sensitivity and that the native and photoactivated forms of rhodopsin can be tuned independently of each other. These results demonstrate the feasibility of designing receptor molecules with specifically modified activated states.

Amino Acid Sequence

Optics of the harbor porpoise eye in water.

A two-dimensional ray-tracing model for the harbor porpoise eye is constructed from new measurements, mainly on two enucleated eyes, and from data found in the literature. Model calculations show that the crystalline lens has too much refractive power to focus light on the retina. The cornea has a high refractive index and acts as a diverging lens of considerable refractive power. The cornea corrects the eye to near emmetropia for axial and temporal (caudal) directions of view. The eye is approximately 5-D myopic for nasal (frontal) directions of view. The iris serves a dual role as a stop: the iris determines the shapes of bundles of light that enter the lens and the iris blocks light that leaves the lens anterior to its equator.

Animals

Enflurane is a potent inhibitor of high conductance Ca(2+)-activated K+ channels of Chara australis.

The volatile anaesthetic, enflurane, is commonly used in surgery for inducing the state of general anaesthesia. It is assumed, that general anaesthetics act on ion channels, but little is known of how they do so and what kinds of channels are sensitive. We found, that enflurane inhibits a large conductance Ca(2+)-activated K+ channel of the green alga, Chara australis. Effects occur at clinically relevant concentrations are fully reversible. The actions of enflurane are distinct from those of charybdotoxin and tetraethylammonium, which are well known blockers of this channel type. Kinetic analysis of single-channel data demonstrates multiple effects of enflurane on the channel protein.

Calcium

Oscillations in the insect brain: do they correspond to the cortical gamma-waves of vertebrates?

gamma-waves, relatively high-frequency oscillations (30-80 Hz) that can be recorded in the olfactory system and the visual cortex of vertebrates, have recently attracted much attention. A role as an information carrier is under discussion, a possible involvement in "feature linking" has been suggested, and they have also been implicated functionally in phenomena such as mind consciousness or awareness. It has long been known that stimulus-dependent high-frequency oscillations (hf waves) can also be recorded from the optic lobes of arthropods. These oscillations in flies have been examined and found to be analogous to the gamma-waves in many respects. Based on knowledge of the anatomy and physiology of the visual system in flies, the most plausible interpretation of the function of these oscillations differs from the interpretations of the vertebrate gamma-waves currently under consideration.

Animals

The nss mutation or lanthanum inhibits light-induced Ca2+ influx into fly photoreceptors.

Ion-selective calcium microelectrodes were inserted into the compound eyes of the wild-type sheep blowfly Lucilia or into the retina of the no steady state (nss) mutant of Lucilia. These electrodes monitored light-induced changes in the extracellular concentration of calcium (delta[Ca2+]o) together with the extracellularly recorded receptor potential. Prolonged dim lights induced a steady reduction in [Ca2+]o during light in the retina of normal Lucilia, while relatively little change in [Ca2+]o was observed in the retina of the nss mutant. Prolonged intense light induced a multiphasic change in [Ca2+]o: the [Ca2+]o signal became transient, reaching a minimum within 6 s after light onset, and then rose to a nearly steady-state phase below the dark concentration. When lights were turned off, a rapid increase in [Ca2+]o was observed, reaching a peak above the dark level and then declining again to the dark level within 1 min. In analogy to similar studies conduced in the honeybee drone, we suggest that the reduction in [Ca2+]o reflects light-induced Ca2+ influx into the photoreceptors, while the subsequent increase in [Ca2+]o reflects the activation of the Na-Ca exchange which extrudes Ca2+ from the cells. In the nss mutant in response to intense prolonged light, the receptor potential declines to baseline during light while the Ca2+ signal is almost abolished, revealing only a short transient reduction in [Ca2+]o. Application of lanthanum (La3+), but not nickel (Ni2+), into the retinal extracellular space of normal Lucilia mimicked the effect of the nss mutation on the receptor potential, while complete elimination of the Ca2+ signal in a reversible manner was observed. The results suggest that La3+ and the nss mutation inhibit light-induced Ca2+ influex into the photoreceptor in a manner similar to the action of the trp mutation in Drosophila, which has been shown to block specifically a light-activated Ca2+ channel necessary to maintain light excitation.

Animals

Ectopic expression of ultraviolet-rhodopsins in the blue photoreceptor cells of Drosophila: visual physiology and photochemistry of transgenic animals.

We have generated transgenic flies expressing R7 cell-specific opsins in the major class of photoreceptor cells of the Drosophila retina and characterized their spectral properties using high-resolution microspectrophotometry and sensitivity recordings. We show that the Rh3 and Rh4 opsin genes encode UV-sensitive opsins with similar spectral properties (lambda max = 345 nm and 375 nm), and that Rh3 corresponds to the R7p and R7marg class of visual pigments. We have also generated Rh3 and Rh4 isoform-specific antibodies and present an R7 cell map of the Drosophila retina. In a related set of experiments, we show that it is possible to coexpress two different visual pigments functionally in the same cell and produce photoreceptors that display the summed spectral response of the individual pigments. These findings open up the possibility of tuning an animal's visual behavior by targeted expression of combinations of opsin genes to selective types of photoreceptors.

Animals

An optomotor control system with automatic compensation for contrast and texture.

When an animal's surroundings move, the animal normally follows that movement by turning its eyes (that is, by an optomotor reaction). As a result, the retinal image is partly stabilized. The efficacy of this stabilization necessarily depends on the gain of the optomotor control circuit. So far no biological detectors of retinal image movements have been discovered in either vertebrates or invertebrates that is, elements capable of generating a signal proportional to the movement velocity, which could serve as sensors in this control system (Borst & Egelhaaf 1989). The reason is that many other parameters, such as the light intensity and the 'texture' of the pattern, also affect the neuronal output. If movement detection is texture dependent, for instance, the gain and hence the quality of stabilization must also be texture dependent. But in humans, at least, with large-field stimulation the quality of retinal image stabilization has been found to be largely independent of texture (de Graaf et al. 1990). Here I describe a control system with gain control that permits automatic compensation, under closed-loop conditions, of the dependence of movement detection on parameters such as texture, brightness and so on. Comparison with data from experiments on arthropods shows that, in these animals at least, a control circuit with nonlinear properties like those suggested here has in fact been realized.

Animals

The pigeon's eye viewed through an ophthalmoscopic microscope: orientation of retinal landmarks and significance of eye movements.

The retina of live, anaesthetized pigeons was inspected with an ophthalmoscopic microscope mounted on a goniometer. Retinal landmarks (optic axis, pecten, fovea, border between the yellow and red field) and the ora terminalis were projected into the visual field of the eye and related to existing data. The resting position of the eye is determined by an orientation of the pecten 45 degrees to the horizontal plane and the optic axis pointing to the horizon with an azimuth angle of 70 degrees relative to the bill. The binocular overlap is maximal (approximately 30 degrees) some 15 degrees above the eye-bill axis. In the resting position of the eye the red field is directed to the lower frontal visual field with only marginal binocular overlap. Binocular overlap of the area dorsalis with the red field, however, during frontal fixation is brought about by eye movements in the range we have demonstrated. The fixation point is 10 degrees below the eye-bill axis.

Animals