Search PubMed⌕ Search

Biomedical subjects

W Denk

Publications and source records attributed to W Denk.

At least 37 records · Page 2Linked to original sources
PubMed1996

Direct measurement of coupling between dendritic spines and shafts.

Characterization of the diffusional and electrotonic coupling of spines to the dendritic shaft is crucial to understanding neuronal integration and synaptic plasticity. Two-photon photobleaching and photorelease of fluorescein dextran were used to generate concentration gradients between spines and shafts in rat CA1 pyramidal neurons. Diffusional reequilibration was monitored with two-photon fluorescence imaging. The time course of reequilibration was exponential, with time constants in the range of 20 to 100 milliseconds, demonstrating chemical compartmentalization on such time scales. These values imply that electrical spine neck resistances are unlikely to exceed 150 megohms and more likely range from 4 to 50 megohms.

Animals↗
PubMed1996

Oscillations and gaseous oxides in invertebrate olfaction.

Olfactory systems combine an extraordinary molecular sensitivity with robust synaptic plasticity. Central neuronal circuits that perform pattern recognition in olfaction typically discriminate between hundreds of molecular species and form associations between odor onsets and behavioral contingencies that can last a lifetime. Two design features in the olfactory system of the terrestrial mollusk Limax maximus may be common elements of olfactory systems that display the twin features of broad molecular sensitivity and rapid odor learning: spatially coherent oscillations in the second-order circuitry that receives sensory input; and involvement of the interneuronal messengers nitric oxide (NO) and carbon monoxide (CO) in sensory responses and circuit dynamics of the oscillating olfactory network. The principal odor processing center in Limax, the procerebrum (PC) of the cerebral ganglion, contains on the order of 10(5) local interneurons and receives both direct and processed input from olfactory receptors. Field potential recordings in the PC show an oscillation at approximately 0.7 Hz that is altered by odor input. Optical recordings of voltage changes in local regions of the PC show waves of depolarization that originate at the distal pole and propagate to the base of the PC. Weak odor stimulation transiently switches PC activity from a propagating mode to a spatially uniform mode. The field potential oscillation in the PC lobe depends on intercellular communication via NO, based on opposing effects of reagents that decrease or increase NO levels in the PC. Inhibition of NO synthase slows the field potential oscillation, while application of exogenous NO increases the oscillation frequency. A role for CO in PC dynamics is suggested by experiments in which CO liberation increases the PC oscillation frequency. These design features of the Limax PC lobe odor processing circuitry may relate to synaptic plasticity that subserves both connection of new receptors throughout the life of the slug and its highly developed odor learning ability.

Animals↗
PubMed1996

Brain lipid peroxidation and hydroxy radical attack following the intravenous infusion of hydrogen peroxide in an infant.

Death following peroxide administration in humans has been reported repeatedly. Hydrogen peroxide, an odorless and clear solution is considered a harmless liquid and is in use for cleaning of superficial wounds. We describe the fatal infusion of this compound by mistake leading to oxygen embolism and, subsequently, to death as a warning for the clinician. Hydrogen peroxide is suggested a major substrate for the in vivo production of the potent oxidizing free radical species "hydroxy radical." No direct evidence for its in vivo production from hydrogen peroxide has been described so far. Using the principle of o-tyrosine determination we studied the formation of the hydroxy radical in the postmortem brain of the infant given intravenous hydrogen peroxide in comparison to postmortem brain samples from five infants. o-Tyrosine is formed by hydroxy radical attack on free and bound phenylalanine and was increased twofold in the brain of the infant given hydrogen peroxide. The significant increase of brain malondialdehyde, a major product and indicator of lipid peroxidation, paralleled the findings of hydroxy radical attack, suggesting that this reactive species has been leading to elevated lipid peroxidation. We propose that the generation of lipid peroxidation and the hydroxy radical from hydrogen peroxide can take place in humans.

Brain↗
PubMed1996

Imaging calcium dynamics in dendritic spines.

Recent advances in optical imaging technology have enabled the measurement of Ca2+ dynamics in individual synaptic spines with high time resolution. Results from work using this new technology have confirmed the view that individual synaptic spines can act as functional chemical compartments with independent dynamics of second-messenger concentration. In particular, the ability of Ca2+ to directly mediate Hebbian coincidence detection has been confirmed.

Animals↗
PubMed1995

Two types of calcium response limited to single spines in cerebellar Purkinje cells.

Of fundamental importance in understanding neuronal function is the unambiguous determination of the smallest unit of neuronal integration. It was recently suggested that a whole dendritic branchlet, including tens of spines, acts as the fundamental unit in terms of dendritic calcium dynamics in Purkinje cells. By contrast, we demonstrate that the smallest such unit is the single spine. The results show, by two-photon excited fluorescence laser scanning microscopy, that individual spines are capable of independent calcium activation. Moreover, two distinct spine populations were distinguished by their opposite response to membrane hyperpolarization. Indeed, in a subpopulation of spines calcium entry can also occur through a pathway other than voltage-gated channels. These findings challenge the assumption of a unique parallel fiber activation mode and prompt a reevaluation of the level of functional complexity ascribed to single neurons.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1995

Dendritic spines as basic functional units of neuronal integration.

Most excitatory synaptic connections occur on dendritic spines. Calcium imaging experiments have suggested that spines constitute individual calcium compartments, but recent results have challenged this idea. Using two-photon microscopy to image fluorescence with high resolution in strongly scattering tissue, we measured calcium dynamics in spines from CA1 pyramidal neurons in slices of rat hippocampus. Subthreshold synaptic stimulation and spontaneous synaptic events produced calcium accumulations that were localized to isolated spines, showed stochastic failure, and were abolished by postsynaptic blockers. Single somatic spikes induced fast-peaking calcium accumulation in spines throughout the cell. Pairing of spikes with synaptic stimulation was frequently cooperative, that is, it resulted in supralinear calcium accumulations. We conclude: (1) calcium channels exist in spine heads; (2) action potentials invade the spines; (3) spines are individual calcium compartments; and (4) spines can individually detect the temporal coincidence of pre- and postsynaptic activity, and thus serve as basic functional units of neuronal integration.

Action Potentials↗
PubMed1995

Calcium imaging of single stereocilia in hair cells: localization of transduction channels at both ends of tip links.

Mechanically gated "transduction" channels in inner ear hair cells are thought to be connected to tip links stretched between adjacent stereocilia. To locate active channels, calcium-green fluorescence in single stereocilia was measured with two-photon laser scanning microscopy. Bundle deflection increased fluorescence in many but not all stereocilia; the increase was blocked by depolarization. The number of stereocilia responding was proportional to the transduction current, consistent with Ca2+ influx through transduction channels. Fluorescence rose first in the tips of stereocilia and then in the bases, in agreement with channel localization at the tips. Some of the shortest stereocilia in a bundle showed a fluorescence increase, as did some of the tallest, indicating that transduction channels can be at either or both ends of tip links.

Animals↗
PubMed1994

Two-photon scanning photochemical microscopy: mapping ligand-gated ion channel distributions.

The locations and densities of ionotropic membrane receptors, which are responsible for receiving synaptic transmission throughout the nervous system, are of prime importance in understanding the function of neural circuits. It is shown that the highly localized liberation of "caged" neurotransmitters by two-photon absorption-mediated photoactivation can be used in conjunction with recording the induced whole-cell current to determine the distribution of ligand-gated ion channels. The technique is potentially sensitive enough to detect individual channels with diffraction-limited spatial resolution. Images of the distribution of nicotinic acetylcholine receptors on cultured BC3H1 cells were obtained using a photoactivatable precursor of the nicotinic agonist carbamoylcholine.

Animals↗
PubMed1994

Anatomical and functional imaging of neurons using 2-photon laser scanning microscopy.

Light scattering by brain tissue and phototoxicity are major obstacles to the use of high-resolution optical imaging and photo-activation ('uncaging') of bioactive compounds from inactive ('caged') precursors in intact and semi-intact nervous systems. Optical methods based on 2-photon excitation promise to reduce these obstacles (Denk, 1994; Denk et al., 1990, 1994). Here we show a range of imaging modes based on 2-photon laser scanning microscopy (TPLSM) as applicable to problems in neuroscience. Fluorescence images were taken of neurons labeled with ion-sensitive and voltage-sensitive dyes in invertebrate ganglia, mammalian brain slices, and from the intact mammalian brain. Scanning photochemical images with whole-cell current detection (Denk, 1994) show how the distribution of neurotransmitter receptors on the surface of specific cells can be mapped. All images show strong optical sectioning and usable images can be obtained at depths greater than 100 microns below the surface of the preparation.

Animals↗
PubMed1994

Assault and homicide by chop blows: a series of 11 cases.

In a series of robberies, 11 elderly women living alone were attacked by an assailant (HS) by means of chop blows directed primarily at the throat, but also at the head, neck, and thorax. 5 died of their injuries. 1 died immediately after the assault. The other 4 victims survived for between 7-38 days. As a result of the injuries incurred, the women were confused, physically weakened and confined to bed, and eventually died from pneumonia. Their death was thus casually related to the violence sustained. Extensive haematomas of the scalp and cervical soft tissue were present. In 3 cases there were injuries to the laryngeal skeleton and adjacent soft tissue. In 2 cases there were conjunctival haemorrhages. The other 6 attacks were not fatal. The blows led to transient unconsciousness, which often resulted in a state of confusion, sometimes mimicking cerebrovascular accidents (CVA). Of other injuries, craniofacial haematomas were the most common. These cases are reviewed in this paper.

Journal Article↗
PubMed1993

Oscillatory flow in the cochlea visualized by a magnetic resonance imaging technique.

We report a magnetic resonance imaging technique that directly measures motion of cochlear fluids. It uses oscillating magnetic field gradients phase-locked to an external stimulus to selectively visualize and quantify oscillatory fluid motion. It is not invasive, and it does not require optical line-of-sight access to the inner ear. It permits the detection of displacements far smaller than the spatial resolution. The method is demonstrated on a phantom and on living rats. It is projected to have applications for auditory research, for the visualization of vocal tract dynamics during speech and singing, and for determination of the spatial distribution of mechanical relaxations in materials.

Animals↗
PubMed1992

Forward and reverse transduction at the limit of sensitivity studied by correlating electrical and mechanical fluctuations in frog saccular hair cells.

The spontaneous fluctuations of the intracellular voltage and the position of the sensory hairbundle were measured concurrently using intracellular microelectrodes and an optical differential micro interferometer. Magnitude and frequency distribution of the hair bundles' spontaneous motion suggest that it consists mostly of Brownian motion. The electrical noise, however, exceeds the value expected for thermal Johnson noise by several orders of magnitude, and its frequency distribution reflects the transduction tuning properties of the hair cells. Frequently, a strong correlation was observed between the fluctuations of the hair bundle position and the intracellular electrical noise. From the properties of the correlation and from experiments involving mechanical stimulation we conclude that in most cases mechano-electrical transduction of the bundles' Brownian motion causes this correlation. Small signal transduction sensitivities ranged from 18 to 500 microV/nm. Bundle motion that was observed in response to current injection in more than half of the cells suggests the existence of a fast reverse (electro-mechanical) transduction mechanism to be common in these cells. The sensitivities could be as high as 600 pm of bundle deflection per millivolt of membrane potential change. In a significant minority (4 in 44) of cells, all showing excess electrical noise, we found 'non-causal' components of the electro-mechanical correlation, and in two of those cells narrow-band bundle motion in excess of their thermal motion at frequencies coincident with peaks in the intracellular noise was observed.

Animals↗
PubMed1992

Mechanical response of frog saccular hair bundles to the aminoglycoside block of mechanoelectrical transduction.

1. Deflections of the mechanosensory hair bundles on frog saccular hair cells were measured interferometrically, with submillisecond temporal and submicrometer spatial resolution, and with subnanometer displacement sensitivity. 2. The direction of the initial bundle deflection (toward the taller stereocilia) in response to a sudden application of aminoglycoside antibiotics shows that the mechanosensory channels are blocked in their mechanically open state. 3. The magnitude of the initial deflection is consistent with published data on the gating swing as derived from the gating compliance. 4. A delayed relaxation and frequently a reversal of the initial deflection were observed and are attributed to the previously reported mechanical adaptation mechanism, which is at least partially controlled by the influx of Ca2+ through the transduction channels. 5. Increases of low-frequency spontaneous motion were found at intermediate blocker concentrations. They can be well accounted for by the fluctuating force exerted on the bundle by the random binding and unbinding of blocker molecules. 6. The mechanical response of the hair bundle to aminoglycosides may be related to their acute and specific ototoxicity.

Aminoglycosides↗
PubMed1992

[Reliability of the Alco-Screen dip strips in evaluation of blood alcohol concentration for clinical or forensic studies].

We tested the accuracy of Alco-Screen dipsticks in comparison to the values of blood alcohol. The dipstick test is based on the reaction of alcohol-dehydrogenase, developing a green to blue color. Saliva as well as urine and liquor are easy to obtain at autopsies. We did not get any false-negative results. Interpretation of the change of color turned out to be a problem, because the tip appeared often to be inhomogenous. The dipstick could be used at autopsies and for emergency units, if a quick, semiquantitative result is required.

Alcohol Drinking↗
PubMed1992

[Medical and technical aspects of weapon effects on man. Part 5--Common weapons].

"Weapons at hand" are tools of everyday life, which can be used as instruments for fighting. The investigation offers a survey concerning instruments, which were actually used for conflicting bodily injuries (cases selected from 1029 expertises) or for killing (cases selected from 474 autopsies). The findings are complemented by non-medical publications about self-defence. In the Austrian legal system the use of "weapons at hand" is interpreted like weapons in the technical sense.

Expert Testimony↗
PubMed1991

[Physician's responsibility in blood transfusions].

Organizational and legal aspects of the blood transfusion are seen under the point of view of division of work. Interpreting the legal directions, people who are involved will be obliged to a duty of care, which is adapted to their position in the operation. A higher degree of responsibility will have to be demanded of the superior, in comparison to his subordinates.

Blood Group Incompatibility↗
PubMed1991

[Medical and technical aspects of weapon effects. IV. Plastic bullets reduce the risk of the military assault rifle (StG 77)].

56 x 45 mm Rota-ammunition was introduced to meet the requirements of security personnel at prisons. The Austrian military assault rifle (StG 77) can be used for this ammunition. The bullet consists of an X-ray dense plastic material with a weight of 1.6 g. Therefore the kinetic energy of this projectile is lower than that of the standard ammunition, also on its flight it loses velocity at a higher rate. Thus, this bullet has the advantage of being less dangerous over long distances (500 m versus 2500 m or more) but still offers at short distances (fe 10 m) a reasonable capacity of incapacitation. Tests concerning ballistics, penetration and shooting at gelatine and soap targets were performed. By means of these tests we compared the wounding capacity of the standard ammunition (5.56 x 45 mm S-cartridge) and the investigated Rota-ammunition.

Animals↗