Ervin Wolf

Persistent responses to brief stimuli: feedback excitation among brainstem neurons

The ability of brief stimuli to trigger prolonged neuronal activity is a fundamental requirement in nervous systems, common to motor responses and short-term memory. Bistable membrane properties and network feedback excitation have both been proposed as suitable mechanisms to sustain such persistent responses. There is now good experimental evidence for membrane bistability. In contrast, the long-standing hypotheses based on positive feedback excitation have yet to be supported by direct evidence for mutual excitatory connections between appropriate neurons. In young frog tadpoles (Xenopus), we show that a small region of caudal hindbrain and rostral spinal cord is sufficient to generate prolonged swimming in response to a brief stimulus. We used paired whole-cell patch recordings to identify hindbrain neurons in this region that actively excite spinal neurons to drive sustained swimming. We show directly that some of these hindbrain neurons make reciprocal excitatory connections with each other. We use a population model of the hindbrain network to illustrate how feedback excitation can provide a robust mechanism to generate persistent responses. Our recordings provide direct evidence for feedback excitation among neurons within a network that drives a prolonged response. Its presence in a lower brain region early in development suggests that it is a basic feature of neuronal network design.

The Extent of the Dendritic Tree and the Number of Synapses in the Frog Motoneuron.

Frog motoneurons were intracellularly labelled with cobaltic lysine in the brachial and the lumbar segments of the spinal cord, and the material was processed for light microscopy in serial sections. With the aid of the neuron reconstruction system NEUTRACE, the dendritic tree of neurons was reconstructed and the length and surface area of dendrites measured. The surface of somata was determined with the prolate – oblate average ellipsoid calculation. Corrections were made for shrinkage and for optical distortion. The mean surface area of somata was 6710 μm2; lumbar motoneurons were slightly larger than brachial motoneurons. The mean length of the combined dendritic tree of brachial neurons was 29 408 μm and that of lumbar neurons 46 806 μm. The mean surface area was 127 335 μm2 in brachial neurons, and 168 063 μm2 in lumbar neurons. The soma – dendrite surface area ratio was 3 – 5% in most cases. Dendrites with a diameter of ≤ 1.0 μm constituted ∼ 75% of the combined dendritic length in most of the neurons. Unlike in the cat, there was no correlation between the size of stem dendrites and the extent of daughter branches. From the synaptic density estimated in earlier electron microscope investigations of frog motoneuron dendrites (Antal et al., J. Neurocytol., 15, 303–310, 1986; 21, 34–49, 1992), and from the present data, the number of synapses on the dendritic tree was calculated. The calculations indicated 26 949 synapses on the smallest and 61 519 synapses on the largest neuron if the synaptic density was multiplied by the length of the dendritic tree. If the synaptic density was multiplied by the surface area of the dendritic tree the calculation yielded 23 337 synapses for the smallest and 60 682 synapses for the largest neuron. More than 60% of the combined surface area of dendrites was >600 μm from the soma. This suggests that about two‐thirds of the synapses impinged upon distant dendrites >600 μm from the soma. The efficacy of synapses at these large distances is investigated on model neurons in the accompanying paper (Wolf et al., Eur. J. Neurosci., 4, 1013–1021, 1992).

A fast 3-dimensional neuronal tree reconstruction system that uses cubic polynomials to estimate dendritic curvature.

The main goal of this work was to develop and test the accuracy of our 3DARBOR neuronal tree reconstruction system by comparing it with a very precise but time-consuming method of reconstruction (NEUTRACE). Comparison was performed by reconstructing 18 dendritic trees of frog spinal motoneurons from serial sections with both methods and comparing several morphological summaries of the two reconstructions. In 3DARBOR the planar projection of the dendritic trees was drawn and fed into an IBM-compatible PC through a graphic tablet. Dendritic coordinates along the perpendicular (focus) axis on the plane of drawing were estimated by an interpolation method. The interpolation was based on the lengths of projected dendrites and the coordinates of points where dendrites entered the next section. Focus axis coordinates of these points could automatically be calculated from the serial numbers and thicknesses of sections. 3DARBOR was tested by comparison of the distributions of characteristic points of dendritic trees, segment lengths and branching angles. Product moment analysis on dendritic trees was also performed. It was concluded that 3DARBOR is a fast enough reconstruction system without any systematical error of interpolation that can correctly supply the most morphological parameters and visualize the natural arborizations.

Simulation of the Effect of Synapses: the Significance of the Dendritic Diameter in Impulse Propagation

The effectiveness of synapses at various sites of the dendritic tree was studied using a segmental cable model with a program developed by Hines (Int. J. Biomed. Comput., 24, 55–68, 1989). The model rendered possible a high‐fidelity simulation of the dendritic geometry of a frog motoneuron described in the accompanying paper (Birinyi et al., Eur. J. Neurosci., 1003–1012, 1992). The model was used in the passive membrane mode and the synaptic activity was simulated with current injections into large and small diameter dendrites at proximal and distal locations. Synaptic efficiency was defined by the charge transfer ratio expressed as the proportion of the injected current which appeared at the soma. The charge transfer ratio was determined with uniform and non‐uniform distribution of specific membrane resistance over the soma–dendrite surface while the diameter of selected dendrite segments changed. The best charge transfer ratio was found with the largest dendrite membrane resistance, and the maximum efficiency of synaptic activity appeared at the original size of the dendrite segment stimulated. The amount of current that flowed in the proximal and distal directions from the segment stimulated depended on the diameter of that segment. The increase in diameter of proximal dendrites increased synaptic efficiency on distal dendrites, whereas the reverse caused a decline in synaptic efficiency on proximal dendrites. In addition to the diameter of dendrites, the arborization pattern also played a significant role in this mechanism. It is concluded that the cellulipetal increase in dendrite diameter greatly increases synaptic efficiency.

The effect of vestibular nerve section on the expression of the hyaluronan in the frog, Rana esculenta

Following postganglionic lesion of the eighth cranial nerve, the changes in the expression of hyaluronan (HA), one of the extracellular matrix macromolecules, were examined in the medial (MVN) and lateral (LVN) vestibular nuclei and in the entry or transitional zone (TZ) of the nerve in the frog. HA was detected in different survival times by using a specific biotinylated hyaluronan-binding probe. HA expression was defined by the area-integrated optical density (AIOD), calculated from pixel intensities of digitally captured images. During the first postoperative days the perineuronal net (PN), a HA-rich area around the neurons, was not distinguishable from the surrounding neuropil in the MVN and LVN, characterized by a bilateral drop of AIOD specifically on the operated side. From postoperative day 14 onwards AIOD increased whilst the PN reorganized. In contrast, the AIOD wobbled up and down bilaterally without any trend in the TZ. Statistical analysis indicated that AIOD changes in the structures studied ran parallel bilaterally presumably because of the operation. Our results demonstrated for the first time that (1) the lesion of the eighth cranial nerve is accompanied by the modification of AIOD reflected HA expression in the MVN, LVN and TZ, (2) different tendencies exist in the time course of AIOD in the structures studied and (3) these tendencies are similar on the intact and operated sides. Our findings may suggest an area dependent molecular mechanism of HA in the restoration of vestibular function.

A Novel Form of Compensation in the Tg2576 Amyloid Mouse Model of Alzheimer’s Disease

One century after its first description, pathology of Alzheimer’s disease (AD) is still poorly understood. Amyloid-related dendritic atrophy and membrane alterations of susceptible brain neurons in AD, and in animal models of AD are widely recognized. However, little effort has been made to study the potential effects of combined morphological and membrane alterations on signal transfer and synaptic integration in neurons that build up affected neural networks in AD. In this study spatial reconstructions and electrophysiological measurements of layer II/III pyramidal neurons of the somatosensory cortex from wild-type (WT) and transgenic (TG) human amyloid precursor protein (hAPP) overexpressing Tg2576 mice were used to build faithful segmental cable models of these neurons. Local synaptic activities were simulated in various points of the dendritic arbors and properties of subthreshold dendritic impulse propagation and predictors of synaptic input pattern recognition ability were quantified and compared in modeled WT and TG neurons. Despite the widespread dendritic degeneration and membrane alterations in mutant mouse neurons, surprisingly little, or no change was detected in steady-state and 50 Hz sinusoidal voltage transfers, current transfers, and local and propagation delays of PSPs traveling along dendrites of TG neurons. Synaptic input pattern recognition ability was also predicted to be unaltered in TG neurons in two different soma-dendritic membrane models investigated. Our simulations predict the way how subthreshold dendritic signaling and pattern recognition are preserved in TG neurons: amyloid-related membrane alterations compensate for the pathological effects that dendritic atrophy has on subthreshold dendritic signal transfer and integration in layer II/III somatosensory neurons of this hAPP mouse model for AD. Since neither propagation of single PSPs nor integration of multiple PSPs (pattern recognition) changes in TG neurons, we conclude that AD-related neuronal hyperexcitability cannot be accounted for by altered subthreshold dendritic signaling in these neurons but hyperexcitability is related to changes in active membrane properties and network connectivity.

Dendritic Impulse Propagation is Altered in Neocortical Pyramidal Neurons of Tg2576 Mice, an Animal Model of Alzheimer's Disease

Alzheimers disease (AD) is one of the most frequent neurological degenerative disorders. During the course of the illness accumulations of amyloid-beta peptide appear in the brain tissue as insoluble extracellular plaques, and neurons and synapses show signs of degeneration. We investigated the dendritic impulse propagation in layer II/III pyramidal neurons of the somatosensory cortex in human amyloid precursor protein over expressing Tg2576 transgenic mice and compared the signal propagation to that of the control, healthy neurons from wild type mice. Earlier studies detected morphological changes in these pyramidal neurons of Tg2576 mice. However, physiological measurements with somatic electrodes could not differentiate between the mutant and control principal neurons based on their passive membrane properties and action potential generation. We used morphologically detailed passive segmental cable models of these cells (n=58) within the NEURON (Duke University, USA) simulation environment. Current was injected to various dendritic points of mutant and healthy neurons to simulate local activity of synapses and current and voltage transfers to the soma were investigated as a function of path distance of injection site from the soma. Spatial distributions of the dendritic surface area and dendritic length were also studied. First in literature, we investigated the dendritic impulse propagation systematically in pyramidal neurons from Tg2576 mice. We found that current transfer is more effective in the apical dendrites of mutant neurons and concluded that this higher effectivity may contribute to the hyperexcitability of mutant neurons, a phenomenon also found in humans with AD.

Somato-dendritic morphology and dendritic signal transfer properties differentiate between fore- and hindlimb innervating motoneurons in the frog Rana esculenta

BackgroundThe location specific motor pattern generation properties of the spinal cord along its rostro-caudal axis have been demonstrated. However, it is still unclear that these differences are due to the different spinal interneuronal networks underlying locomotions or there are also segmental differences in motoneurons innervating different limbs. Frogs use their fore- and hindlimbs differently during jumping and swimming. Therefore we hypothesized that limb innervating motoneurons, located in the cervical and lumbar spinal cord, are different in their morphology and dendritic signal transfer properties. The test of this hypothesis what we report here.ResultsDiscriminant analysis classified segmental origin of the intracellularly labeled and three-dimensionally reconstructed motoneurons 100% correctly based on twelve morphological variables. Somata of lumbar motoneurons were rounder; the dendrites had bigger total length, more branches with higher branching orders and different spatial distributions of branch points. The ventro-medial extent of cervical dendrites was bigger than in lumbar motoneurons. Computational models of the motoneurons showed that dendritic signal transfer properties were also different in the two groups of motoneurons. Whether log attenuations were higher or lower in cervical than in lumbar motoneurons depended on the proximity of dendritic input to the soma. To investigate dendritic voltage and current transfer properties imposed by dendritic architecture rather than by neuronal size we used standardized distributions of transfer variables. We introduced a novel combination of cluster analysis and homogeneity indexes to quantify segmental segregation tendencies of motoneurons based on their dendritic transfer properties. A segregation tendency of cervical and lumbar motoneurons was detected by the rates of steady-state and transient voltage-amplitude transfers from dendrites to soma at all levels of synaptic background activities, modeled by varying the specific dendritic membrane resistance. On the other hand no segregation was observed by the steady-state current transfer except under high background activity.ConclusionsWe found size-dependent and size-independent differences in morphology and electrical structure of the limb moving motoneurons based on their spinal segmental location in frogs. Location specificity of locomotor networks is therefore partly due to segmental differences in motoneurons driving fore-, and hindlimbs.