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Biomedical subjects

G D Lange

Publications and source records attributed to G D Lange.

35 records · Page 2Linked to original sources
PubMed1989

A fractal analysis of cell images.

Methods of digital image analysis have been adapted to measure the fractal dimension of cellular profiles. The fractal dimension is suggested as a useful measure of the complexity of a contour. Three methods produce similar results when applied to constructed, near-ideal fractal figures. Comparison of the measurements for a variety of image types indicates the measurement accuracy in each case and may help in interpreting the results when applied to real, non-ideal cell images of unknown fractal dimension. Two of the methods are currently adopted as appropriate for use on neuronal contours. A correlation exists between the complexity of these contours and the magnitude of the estimated fractal dimension.

Animals↗
PubMed1988

(-)-Baclofen activates presynaptic GABAB receptors on GABAergic inhibitory neurons from embryonic rat hippocampus.

GABA-mediated monosynaptic inhibitory postsynaptic currents were recorded at synapses between embryonic rat hippocampal neurons in culture. The GABAB receptor agonist L(-)-baclofen reduced inhibitory postsynaptic current amplitude substantially at 10 microM, while the D(+)-isomer of baclofen was without effect. L(-)-Baclofen did not increase postsynaptic membrane conductance or block postsynaptic membrane responses to GABA. It is concluded that L(-)-baclofen depresses GABA-mediated inhibitory postsynaptic potentials by a presynaptic action at GABAB receptors on or near the terminals of the inhibitory neuron. The possible physiological importance of these presynaptic 'autoreceptors' is discussed.

Action Potentials↗
PubMed1988

Flow cytometric analysis of membrane potential in embryonic rat spinal cord cells.

Flow cytometric analysis of membrane potential in suspensions of embryonic rat spinal cord cells was carried out in a fluorescence-activated cell sorter (FACS) using anionic voltage-sensitive, fluorescent dyes (oxonols). The FACS or flow cytometer is an analytical instrument that measures optical properties of large cell populations at a single cell level of resolution. The incorporation of oxonol allows relative measurements of membrane potential, since the partition of oxonol within the plasmalemma is directly related to the degree of cell depolarization. Incubation of cells in elevated K+ concentrations or with the Na+ channel agonist batrachotoxin (BTX) changed the fluorescence intensity distribution pattern of the live-cell population; these changes were consistent with the depolarizing effects of these manipulations. Fluorescence shifts were either undetectable or minimal in the dead-cell population. The BTX-induced shift was blocked by tetrodotoxin (TTX) and was reversed in Na+-free medium, indicating embryonic expression of functional Na+ channels. Fluorescence microscopy of sorted cells showed that live cells typically exhibited circumferential ring-like patterns, whose intensities were enhanced under depolarizing conditions. The results show that flow cytometry combined with oxonol dyes can be used to measure the relative membrane potential of large numbers of individual central nervous system cells. The analysis of the changes in the distributions of these membrane potentials can be used to reveal the development of functional ion conductance mechanisms.

Animals↗
PubMed1988

Edge detection in images using Marr-Hildreth filtering techniques.

Details of the morphology of light microscope images of horseradish peroxidase labeled mammalian neurons in cell culture were investigated. A modified Marr-Hildreth edge-detecting algorithm was used in an image processor to obtain a continuous border of the labeled neurons. The interior of the border was filled to obtain isolated binary silhouettes of the neurons. These silhouettes can be used for further quantitative studies.

Animals↗
PubMed1987

Potentiation of gamma-aminobutyric-acid-activated chloride conductance by a steroid anaesthetic in cultured rat spinal neurones.

1. Intracellular recordings from cultured rat spinal cord neurones demonstrated that Cl(-)-dependent responses to GABA (gamma-aminobutyric acid) (but not glycine) were increased in amplitude and duration by the steroid anaesthetic alphaxalone (3 alpha-hydroxy-5 alpha-pregnane-11,20-dione) at submicromolar concentrations that produced little or no effect on passive electrical properties. The non-anaesthetic 3 beta-hydroxy analogue was without effect on GABA-evoked responses. 2. Under voltage clamp, membrane currents evoked by GABA were potentiated by alphaxalone without change in the reversal potential for the GABA-evoked response. Fluctuation analysis of GABA-evoked currents suggested that the mean open-time of GABA-activated channels was prolonged from 30 to 74 ms in the presence of the anaesthetic. 3. Higher concentrations of alphaxalone, similar to those reported during surgical anaesthesia, increased membrane conductance in the absence of exogenously applied GABA. Under voltage clamp, current responses to alphaxalone reversed at the same potential as did responses to GABA, suggesting that they result from increased Cl- conductance. 4. Alphaxalone responses were reduced by the GABA antagonist bicuculline. Fluctuation analysis of current responses to the anaesthetic suggest that they result from the activation of ion channels of long (100 ms) open-time and elementary conductance indistinguishable from that of channels activated by GABA (20 pS). Taken together, these findings indicate that the steroid anaesthetic is able to directly activate Cl- conductance normally activated by GABA in spinal neurones. 5. The actions of the steroid at GABA-receptor-Cl(-)-channel complexes are similar to those produced by the anaesthetic barbiturates (e.g. pentobarbitone), although obtained at 50-100-fold lower concentrations. These effects on the inhibitory Cl(-)-conductance mechanism may be partly responsible for the depressant actions of alphaxalone on the mammalian central nervous system.

Anesthetics↗
PubMed1987

The influence of skeletal muscle on the electrical excitability of dorsal root ganglion neurons in culture.

Dorsal root ganglion (DRG) neurons from embryonic mice grown in coculture with dissociated skeletal muscle or in skeletal muscle conditioned medium (CM) showed an increased incidence of repetitive firing of action potentials when injected with sustained (60-100 msec) depolarizing current. This is in contrast to DRG neurons grown in monoculture and normal medium, which exhibit such behavior far less frequently. The first action potential showed less sensitivity to block with TTX and more sensitivity to Ca2+ channel blockers than the subsequent action potentials. The increased incidence of repetitive firing occurred when CM was added after as few as 2 or as many as 22 d in culture and with as little as 1-7 hr exposure to CM. This effect of CM cannot be mimicked by NGF or by coculture with cells from embryonic spinal cord (Peacock et al., 1973), can be eliminated by heating the CM at 56 degrees C for 30 min, and partially reversed following short exposure to CM. These results indicate that skeletal muscle releases some heat-labile factor(s) that can cause repetitive firing and, in addition, significant decrease in input resistance in the CM-treated neurons and a depression of the anomalous rectification, neither of which could account for the increase in repetitive firing.

Action Potentials↗
PubMed1986

Analysis and isolation of embryonic mammalian neurons by fluorescence-activated cell sorting.

Cells were dissociated from the CNS of the embryonic mouse and rat to produce cell suspensions suitable for analysis and separation on a fluorescence-activated cell sorter (FACS). Cells from the spinal cord of the embryonic mouse were analyzed in the most detail. Cell suspensions generated three major peaks in histograms of forward-angle light scatter. Examination of material isolated from each peak and labeling of cell suspensions with the nonvital and supravital fluorescent dyes propidium iodide, ethidium bromide, and acridine orange demonstrated that the three peaks represented live cells, dead cells, and subcellular fragments. Passage through the cell sorter did not detectably damage live cells, as shown by light microscopy, FACS analysis, and in vitro culture of sorted cells. Neurons and glial cells collected by sorting survived at least 4 weeks in culture. Cell suspensions dissociated from the dorsal root ganglia, hippocampus, hypothalamus, cerebellum, and cerebral cortex of the embryonic mouse and from the spinal cord of the embryonic rat produced similar results. Analysis of samples prepared at different developmental stages showed that viable cells could be recovered from each of these regions throughout the important stages of neurogenesis and early cellular differentiation, but that few viable cells could be recovered from animals beyond late embryonic or early postnatal ages. Quantitative FACS analysis of monoclonal antibody A2B5, tetanus toxin and cholera toxin, and lectins binding to live dissociated cells from the embryonic spinal cord demonstrated that these cells had already developed binding sites for these cell-surface ligands by embryonic day 13. These results demonstrate that a fluorescence-activated cell sorter can be used for quantitative analysis of specific cellular properties, that FACS analysis and sorting can be used to identify and isolate live cells from many regions of the embryonic mammalian CNS during important developmental periods, and that sorted neurons and glial cells can be maintained for weeks in culture.

Animals↗
PubMed1980

Electroreception in the ratfish (Hydrolagus colliei).

Behavioral and neurophysiological experiments and anatomical work indicate that the ampullar structures on the head of fish of the subclass Holocephali are sense organs responsive to weak electric fields and are functionally and structurally homologous to the ampullae of Lorenzini in elasmobranchs. It is concluded that, as in elasmobranchs, these organs are used to detect bioelectric and other natural electric phenomena in the environment.

Action Potentials↗
PubMed1979

Fourier analysis of spike train data.

A formal representation of nerve spike trains in the form of a sum of rectangular functions is proposed. This formal instantaneous frequency function can be Fourier analyzed. The resulting algorithm has the useful properties of spike by spike calculations and an insensitivity to the mean (carrier) spike rate. The technique is also useful for producing a smooth (filtered) reconstruction of a spike train.

Action Potentials↗
PubMed1977

Sinusoidal analysis of electroretinogram of squid and octopus.

1. An isolated eye or eye plus optic lobe preparation (in oxygenated chilled seawater) from Loligo opalescens, Octopus bimaculata, and O. bimaculoides was used to study the electroretinogram (ERG) for small signal intensity-modulated stationary spots of light. 2. If light intensity was modulated sinusoidally (modulation depth 0-50%) the ERG response is sinusoidal with less than 2% of the power present in the next five harmonics compared to the fundamental. Bode plots, amplitude and phase shift plotted against frequency, were constructed from these sinusoidal input-output experiments. 3. Linearity and time invariance were tested: a) an increase in amplitude of sinusoidal modulation by a constant factor caused an increase in response amplitude by the same factor but caused no change in shape of the Bode plot gain or phase curves; b) the transfer function represented by the Bode plot could be used to predict waveshape of the response to a brief flash (Green's or impulse-response function); c) the Fourier transformed square-wave response could be used to obtain a Bode plot which coincided with that obtained by sinusoidal input-output experiments. 4. The Bode plot can be fit by the transfer function of 5-12 (depending on conditions and on the preparations) series cascaded low-pass filters whose corner frequences are distributed between 0.2 and 40 Hz. Alternatively, 3-7 filters plus a delay of 25-130 ms fits the Bode plots equally well. The series filter model is compatible with a simply physical model consisting of cascaded chemical reactions whose forward rate constants are reciprocals of the filter time constants, whose reverse rate constants are negligible, and in which the concentration of an intermediate product controls membrane current. 5. As mean intensity is increased, the gain decreases. This effect is more pronounced at low frequencies than at high frequencies. Thus, the system is nonlinear for large intensity changes. The process of adaptation involves not only a change in gain, but a change in shape of the Bode plot, i.e., change in filter corner frequencies. In terms of the reaction chain model, this means that some rate constants change as the state of adaptation is changed.

Animals↗
PubMed1975

The influence of dental plaques on the microvasculature of the oral mucous membrane.

This investigation was designed to explore the effect of plaque, obtained at different stages of accumulation, on the microvasculature of the hamster cheek pouch. Five groups of three dental students followed an oral hygiene abstinence program for 22 days. At day 2, 9, and 22, plaque was sampled from different anterior teeth, pooled and suspended in a reduced diluting fluid. Each of the three pooled plaque suspensions was then introduced into the right or left cheek pouche of five hamsters. Microvascular changes in the pouches were then observed and recorded photomicrographically until vascular reconstitution occurred. Plaque and gingival indices were taken, and the microbial composition of the plaque was determined according to gram-staining characteristics and morphology. The results indicate that the microvascular response was not a function of age and composition of plaque. This finding is based on the observation that 2-, 9-, and 22-day-old plaque induced similar microvascular damage which required approximately the same recovery period of 9 days to reconsitute normal microvascular function.

Adult↗
PubMed1974

A nonlinearity in the inhibitory interactions in the lateral eye of limulus.

Receptor units in the eye of the horseshoe crab are more sensitive to lateral inhibition at some levels of excitation than they are at others. As a result, the steady-state inhibition of the response of a given unit is not directly proportional to the response levels of neighboring units. This effect may be represented by the introduction of a nonlinearity in the Hartline-Ratliff system of equations. The nonlinear inhibitory effect appears to increase the operating range of the receptor units.

Animals↗
PubMed1969

Objective measure of the dynamics of a visual movement illusion.

Apparent movement in peripheral vision can be induced by sequential flashing of two dots that are spatially unresolved. Subjects used this illusion to make forced-choice estimates of the directional sequence of the two dots. Performance at this task defines spatiotemporal conditions that induce the illusion without reliance upon subjective distinctions of "movement" from "successivity" and "simultaneity." The dynamics of the illusion, defined in this way, are measured and compared with those for after-flash inhibition and the perception of real movement.

Eye Movements↗