Immunochemical and electrophysiological analyses of magnetically responsive neurons in the mollusc Tritonia diomedea

Tritonia diomedea uses the Earth’s magnetic field as an orientation cue, but little is known about the neural mechanisms that underlie magnetic orientation behavior in this or other animals. Six large, individually identifiable neurons in the brain of Tritonia (left and right Pd5, Pd6, Pd7) are known to respond with altered electrical activity to changes in earth-strength magnetic fields. In this study we used immunochemical, electrophysiological, and neuroanatomical techniques to investigate the function of the Pd5 neurons, the largest magnetically responsive cells. Immunocytochemical studies localized TPeps, neuropeptides isolated from Pd5, to dense-cored vesicles within the Pd5 somata and within neurites adjacent to ciliated foot epithelial cells. Anatomical analyses revealed that neurites from Pd5 are located within nerves innervating the ipsilateral foot and body wall. These results imply that Pd5 project to the foot and regulate ciliary beating through paracrine release. Electrophysiological recordings indicated that, although both LPd5 and RPd5 responded to the same magnetic stimuli, the pattern of spiking in the two cells differed. Given that TPeps increase ciliary beating and Tritonia locomotes using pedal cilia, our results are consistent with the hypothesis that Pd5 neurons control or modulate the ciliary activity involved in crawling during orientation behavior.     

Integration of Sensory Quanta in Cuneate Nucleus Neurons In Vivo

Discriminative touch relies on afferent information carried to the central nervous system by action potentials (spikes) in ensembles of primary afferents bundled in peripheral nerves. These sensory quanta are first processed by the cuneate nucleus before the afferent information is transmitted to brain networks serving specific perceptual and sensorimotor functions. Here we report data on the integration of primary afferent synaptic inputs obtained with in vivo whole cell patch clamp recordings from the neurons of this nucleus. We find that the synaptic integration in individual cuneate neurons is dominated by 4–8 primary afferent inputs with large synaptic weights. In a simulation we show that the arrangement with a low number of primary afferent inputs can maximize transfer over the cuneate nucleus of information encoded in the spatiotemporal patterns of spikes generated when a human fingertip contact objects. Hence, the observed distributions of synaptic weights support high fidelity transfer of signals from ensembles of tactile afferents. Various anatomical estimates suggest that a cuneate neuron may receive hundreds of primary afferents rather than 4–8. Therefore, we discuss the possibility that adaptation of synaptic weight distribution, possibly involving silent synapses, may function to maximize information transfer in somatosensory pathways.     

The sensory functions of the cerebellum of the thornback ray,Platyrhinoidis triseriata

Summary Single unit spikes and evoked field potentials were recorded in different parts and depths of the corpus cerebelli and auricle of immobilized rays before and after stimulating with light, electric fields, touch, tail bending and direct shock to mechanoreceptive nerves of the lateral line.Discrete areas of the cerebellum are responsive to these modalities and the areas show limited overlap; they are all distinct from the area reported by Plassmann to be responsive to angular acceleration. The visual and tactile-proprioceptive areas are large; the electric area is small. Most units are excited only by one modality.The tail is represented only in the posterior lobe; trigeminal innervation extends from the posterior onto the anterior lobe, suggesting some topographic projection.The dynamic characteristics of the responses were examined particulary in the visual units. To a flash, units discharge up to six bursts of spikes in 500 ms. This pattern is reduced at repetition rates > 1/s; above ca. 4/s units tend to fire irregularly. Various kinds of units are found in respect to the succession of responses to short trains of flashes. Some units fire much better to objects moving in a limited visual field with a certain direction and rate.Abbreviation EP evoked potential     

Expression of mRNAs for Neurotrophins (NGF,BDNF, and NT-3) and their Receptors (p75NGFR,Trk, TrkB,and TrkC) in Human Peripheral Neuropathies

The steady-state mRNA levels of NGF, BDNF and NT-3, and the mRNA levels of their receptors p75^NGFR, trk, trk,B and trkC were examined in various human peripheral neuropathies, to determine the correlation with myelinated fiber pathology and T cell and macrophage invasions in the diseased nerves. Steady state levels of p75^NGFR mRNAs were significantly elevated in nerves with axonal pathology. In contrast, steady state levels of trkB and trkC mRNA levels were diminished, trk mRNA was not detected in the human nerves. The NGF, BDNF, and NT-3 mRNA levels were elevated in the diseased nerves. The increase in BDNF and NT-3 mRNA levels were proportional to the extent of invasion of the nerves by T cells and macrophages, but did not directly correlate with axonal nor demyelinating pathology, thus suggesting that inflammatory cell invasions are involved in the regulation of BDNF and NT-3 mRNA expressions. These neurotrophin and their receptor gene expressions in the diseased human nerves would be regulated by an underlying pathology-related process, and could play a role in peripheral nerve repair.     

Changes in lectin binding of lumbar dorsal root ganglia neurons and peripheral axons after sciatic and spinal nerve injury in the rat

Summary The effects of chronic lesions of rat lumbar spinal or sciatic nerves on the binding of Glycine max (soybean) agglutinin to galacto-conjugates, in small-and medium-size primary sensory neurons of the L₄ and L₅ dorsal root ganglia, were examined over a 580-day period. Spinal nerve section resulted in a marked decrease in the population of stained neurons within 7 days. However, despite some retrograde morphological changes triggered by axonal injury, the proportion of stained nerve cells was normalized 180 days postoperatively. This temporary decrease in perikaryal lectin reactivity was initially associated with a marked accumulation of stained material in the nerve, proximal and distal to the site of section, with similar accumulations also being noticeable at each level of injury in sciatic nerves subjected to double ligature. This may reflect the presence of glycocompounds linked to the autolysis of nerve fibers during the phase of retrograde dying-back and Wallerian degeneration. At later stages, stained deposits could be seen scattered along central and peripheral axonal processes of the dorsal root ganglion neurons in the vicinity of the cell body. They may indicate a disturbance in the peripheral turnover of glycoproteins in chronically-transected nerves, with piling up of neuronal products. Sciatic nerve injury caused similar but less severe effects which, except for the L₄ ganglion cells, were rapidly reversible.     

Microelectrode analysis of light responses in the brain of the cricket (Gryllus domesticus)

Electrical activity in response to light stimuli was recorded from the brain of the cricket (Gryllus domesticus) using stainless steel microelectrodes. Four basic types of elements were observed as follows: (1) units which registered ambient light intensity by frequency of firing as well as responding with transient changes in rate to stepwise increases or decreases in intensity; (2) units which fired at a higher frequency in dark than in light; (3) units which fired continuously at low level in light and responded with a transient high frequency burst to light off; and (4) units which responded with a brief burst to on and off, but tended to be “on-dominant” or “off-dominant.” Also observed were synchronized spikes in mushroom body responding primarily to light off, but also on occasion to light on, and often accompanied by single unit responses. The units registering intensity are probably homologous with units showing similar properties recorded from the visual systems of several other arthropods and usually referred to as “sustaining units.” On-off, off, and dark units are also known from other forms. The mushroom body light responses were similar to synchronized spikes recorded in cockroach mushroom body following antennal stimulation.     

Neuronal organization of the periamygdaloid cortex in the cat brain

Neuronal organization of the fields Pmm, Pml2, Pe and epm of the periamygdaloid cortex of the cat brain has been studied by means of Golgi and Nissl methods. The field Pmm essentially differs from other fields of this cortex by primitiveness of its cytoarchitectonic an neuronal organization (two layers uniform by the composition of their neurons are distinguished, the structure of the latter is relatively primitive). In the medial part of this field long axonal rarely branching short dendritic, and in the lateral part--poorly differentiating pyramidal and spindle-like cells predominate. The field Pmm can be considered as a transitional formation between the subcortex (the medial nucleus of the amygdaloid body) and other fields of the periamygdaloid cortex. The fields Pml2, Pe and epm are built more complexly: the cells are organized in 4 layers, more complexly differentiated by their form and size than in the field Pmm and correspondingly more various (long axonal densely branching cells are observed: pyramidal and spindle-like--of the cortical type and bushy--of the subcortical type, as well as long axonal rarely branching reticular cells). The short axonal cells in the fields Pml2, Pe and epm are rather variable in their form, size and direction of axons; in the field Pmm they are less numerous. The field Pmm and the complex of the fields Pml2, Pe and epm are perhaps different in their function, this is evident from different projection of their neurons. Axons of the cells in the field Pmm get into less differentiated and the most ancient medial nucleus of the amygdaloid body and into the ancient system of connections of the latter--terminal strip, and neurons of the fields Pml2, Pe and epm are projected into the basolateral part of the amygdaloid body and into the external capsule--phylogenetically younger structures. Besides, poverty of the axonal collateralies in the long axonal neurons and a small amount and uniformity of the forms of the short axonal cells in the field Pmm and contrary, rich collateralies and variety of short axonal cells in the composition of other fields demonstrate more complex internal integrative function, performing in that composition.     

Ambient 60-Hz magnetic flux density in an urban neighborhood

A residential neighborhood in Buffalo, NY, was surveyed with a magnetic field meter to evaluate whether or not spot measurements are reliable predictors of the 60-Hz fields at street corners and residences. The results of repeated measurements over 7 days at 33 street corners in this neighborhood indicate that day-to-day variation in power line magnetic fields is negligible (intraclass correlation coefficient = 0.94). Multivariate linear regression analysis of the data indicates that transmission lines and thick, three-phase primary wires near the field measurement site are strong predictors and account for the majority of the ambient magnetic field variance between locations (multiple correlation coefficient squared = 0.60; F ratio = 22.2, P less than .001). Magnetic fields measured at the front sidewalk were highly correlated with fields at the front doorsteps of 45 homes in this neighborhood (gamma = 0.81). These results suggest that ambient power line magnetic field levels at urban residences can be reliably characterized on a one-time site inspection using a hand-held magnetic field meter and a simple wiring classification system.     

Magnetic intensity affects cryptochrome-dependent responses in Arabidopsis thaliana

Cryptochromes are blue-light absorbing photoreceptors found in many organisms where they have been involved in numerous growth, developmental, and circadian responses. In Arabidopsis thaliana, two cryptochromes, CRY1 and CRY2, mediate several blue-light-dependent responses including hypocotyl growth inhibition. Our study shows that an increase in the intensity of the ambient magnetic field from 33–44 to 500 μT enhanced growth inhibition in A. thaliana under blue light, when cryptochromes are the mediating photoreceptor, but not under red light when the mediating receptors are phytochromes, or in total darkness. Hypocotyl growth of Arabidopsis mutants lacking cryptochromes was unaffected by the increase in magnetic intensity. Additional cryptochrome-dependent responses, such as blue-light-dependent anthocyanin accumulation and blue-light-dependent degradation of CRY2 protein, were also enhanced at the higher magnetic intensity. These findings show that higher plants are sensitive to the magnetic field in responses that are linked to cryptochrome-dependent signaling pathways. Because cryptochromes form radical pairs after photoexcitation, our results can best be explained by the radical-pair model. Recent evidence indicates that the magnetic compass of birds involves a radical pair mechanism, and cryptochrome is a likely candidate for the avian magnetoreception molecule. Our findings thus suggest intriguing parallels in magnetoreception of animals and plants that appear to be based on common physical properties of photoexcited cryptochromes.     

Bioelectric properties of frog sciatic nerves during exposure to stationary magnetic fields

The bioelectric properties of frog sciatic nerves have been measured during exposure to homogeneous, stationary magnetic fields. The action potential amplitude, conduction velocity, absolute refractory period and relative refractory period were found to be unaffected by a continuous 4-h exposure to perpendicular or parallel 2.0 T (1 T equal 10(4) G) magnetic fields. These parameters also remained unchanged during a 1-h post-exposure period. The conduction velocity was similarly found to be unchanged when the field was applied continuously for 17 h. Exposure of sciatic nerves to a 1.0-T field led to no alteration in the threshold for neural excitation. The absence of magnetic field effects on nerve electrical activity observed in the present experiments contrasts with the positive findings reported previously by other investigators. These discrepancies may be attributable to an inadequate control of ambient temperature in the earlier studies.     

The effects of temperature on signalling in ocellar neurons of the desert locust, <Emphasis Type="Italic">Schistocerca gregaria</Emphasis>

In Schistocerca gregaria ocellar pathways, large second-order L-neurons use graded potentials to communicate signals from the ocellar retina to third-order neurons in the protocerebrum. A third-order neuron, DNI, converts graded potentials into axonal spikes that have been shown in experiments at room temperature to be sparse and precisely timed. I investigated effects of temperature changes that a locust normally experiences on these signals. With increased temperature, response latency decreases and frequency responses of the neurons increase. Both the graded potential responses in the two types of neuron and the spikes in DNI report greater detail about a fluctuating light stimulus. Over a rise from 22 to 35°C the power spectrum of the L-neuron response encompasses higher frequencies and its information capacity increases from about 600 to 1,700 bits/s. DNI generates spikes more often during a repeated stimulus but at all temperatures it reports rapid decreases in light rather than providing a continual measure of light intensity. Information rate carried by spike trains increases from about 50 to 185 bits/s. At warmer temperatures, increased performance by ocellar interneurons may contribute to improved aerobatic performance by delivering spikes earlier and in response to smaller, faster light stimuli.     

Cellular migration and axonal outgrowth from adult mammalian peripheral nerves in vitro

It is known that following peripheral nerve transections, sheath cells proliferate and migrate to form a bridge between nerve stumps, which may facilitate axonal regeneration. In the present investigations, cellular migration and axonal outgrowth from nerves of adult mice were studied in vitro using collagen gels. During the first 3 days in culture, profuse migration of fibroblasts and macrophages occurred from the ends of sciatic nerve segments, which had been lesioned in situ a few days prior to explanation, but not from segments of normal nerves. The mechanism of cellular activation in the lesioned nerves was not determined, but migration was blocked by suramin, which inhibits the actions of several growth factors. The migrating cells, which form the bridge tissue, may promote axonal regeneration in two ways. Firstly, axonal outgrowth from isolated intercostal nerves was significantly increased in co-cultures with bridges from lesioned sciatic nerves. This stimulatory effect was inhibited by antibodies to 2.5S nerve growth factor. Secondly, the segments of bridge tissue contracted when removed from animals. It is possible that fibroblasts within the bridge exert traction that would tend to pull the lesioned stumps of peripheral nerve together, as in the healing of skin wounds. The traction may also influence deposition of extracellular matrix materials, such as collagen fibrils, which could orient the growth of the regenerating axons toward the distal nerve stump. © 1996 John Wiley & Sons, Inc.     

Axonal branching pattern and coupling mechanisms of the cerebral giant neurones in the snail,lymnaea stagnalis

The axonal branching pattern of the two cerebral giant neurones (CGCs) of Lymnaea stagnalis was studied with intrasomatically applied horseradish peroxidase. The cells are symmetrical. Each CGC projects to the ipsilateral n. labialis medius and n. arteriae labialis, the subcerebral commissure, and to all ipsi- and contralateral buccal nerves. The contralateral buccal nerves are reached via the ipsilateral cerebro-buccal connective and the buccal commissure. The CGC fire action potentials 1:1 in a driver-follower relationship. Each cell is capable of both driving and following. The relationship depends on the membrane potentials of the somata. In driving CGC spikes are initiated in a cerebral spike trigger zone located near the soma. In following cells spikes are initiated in a distal zone located in the buccal ganglia. The buccal zone is only affected by the partner CGC. CGC are synchronized by three coupling mechanisms: mutual excitatory chemical synapses, electrotonic coupling, and common input. The chemical and electrotonic connections are located in the buccal ganglia. All spikes are relayed to the partner cell via the chemical synapses. The electrotonic coupling improves the efficiency of the chemical synapses. The dual connection selectively synchronizes the CGC-axonal spikes from each side of the buccal mass. Common excitatory input affects the cerebral spike trigger zones and can initiate simultaneous spikes in both cells. This results in bilateral synchrony of spikes in the CGC-axons in both the buccal and the lip nerves.     

β2-Adrenergic Receptors on Peripheral Nerves

We report that peripheral nerves have a functional adenylate cyclase-coupled beta-adrenergic receptor. The pharmacological specificity of this receptor is shown to be of the beta 2 subtype. Two peripheral nerves, the sciatic from the frog and rat and the vagus from the rat, responded to beta 2-agonists with 10-50-fold increases in intracellular cyclic AMP level. This increase was inhibited by the beta-adrenergic antagonist propranolol. In contrast, a central nerve tract, the corpus callosum, responded to isoproterenol with only a minimal one- to twofold increase in cyclic AMP level. These studies demonstrate that peripheral nerves have beta 2-adrenergic receptors that are responsive to exogenously applied catecholamines and suggest a role for these ligands in the previously described modulation of axonal conduction.     

Comparison of coupling of humans to electric and magnetic fields with frequencies between 100 Hz and 100 kHz

Recent laboratory and epidemiological results have stimulated interest in the hypothesis that human beings may exhibit biological responses to magnetic and/or electric field transients with frequencies in the range between 100 Hz and 100 kHz. Much can be learned about the response of a system to a transient stimulation by understanding its response to sinusoidal disturbances over the entire frequency range of interest. Thus, the main effort of this paper was to compare the strengths of the electric fields induced in homogeneous ellipsoidal models by uniform 100 Hz through 100 kHz electric and magnetic fields. Over this frequency range, external electric fields of about 25–2000 V/m (depending primarily on the orientation of the body relative to the field) are required to induce electric fields inside models of adults and children that are similar in strength to those induced by an external 1 μT magnetic field. Additional analysis indicates that electric fields induced by uniform external electric and magnetic fields and by the nonuniform electric and magnetic fields produced by idealized point sources will not differ by more than a factor of two until the sources are brought close to the body. Published data on electric and magnetic field transients in residential environments indicate that, for most field orientations, the magnetic component will induce stronger electric fields inside adults and children than the electric component. This conclusion is also true for the currents induced in humans by typical levels of 60 Hz electric and magnetic fields in U.S. residences. Bioelectromagnetics 18:67–76, 1997. © 1997 Wiley-Liss, Inc.     

Organization of the Viscerosomatic Afferent Projections to Fields 24 and 25 of the Cingulate Gyrus of the Cat Brain Cortex

We studied evoked potentials (EP) and responses of single neurons in fields 24 and 25 of the cingulate gyrus of the cat brain cortex. The responses were elicited by stimulations of a parasympathetic, a sympathetic, and a somatic nerve (the pelvic, splanchnic, and sciatic nerves). The configuration and amplitude/temporal characteristics of both EP and neuronal responses showed that the organization of the visceral and somatic afferent inputs to the cortical fields under study is to a great extent similar. Localization of the foci of maximum activity for EP within field 24 and their overlapping, the higher responsiveness of the neurons of this field to afferent volleys as compared with that in field 25, the dominance of short-latency neuronal responses in field 24, and the presence of polysensory neurons with modally specific opposite responses in this field allow us to conclude that field 24 (compared with field 25) plays a more significant role in the perception and analysis of viscerosomatic afferent information. Taking into account earlier published data that the zones of concentration of the efferent neurons (forming a descending limbico-sympathetic discharge in the L2-L3 white connective branches and a limbico-parasympathetic discharge in the pelvic nerve) are localized within field 25 and overlap each other, we discuss the functional roles of fields 24 and 25 of the cingulate gyrus in viscerosensory and visceromotor control.     

Pedal neuron 3 serves a significant role in effecting turning during crawling by the marine slug <Emphasis Type="Italic">Tritonia diomedea</Emphasis> (Bergh)

The marine nudibranch Tritonia diomedea crawls using its ciliated foot surface as the sole means of propulsion. Turning while crawling involves raising a small portion of the lateral foot margin on the side of the turn. The cilia in the lifted area no longer contribute to propulsion, and this asymmetry in thrust turns the animal towards the lifted side. Neurons located in the pedal ganglia of the brain contribute to these foot margin contractions. T. diomedea has a natural tendency to turn upstream (rheotaxis), and pedal flexion neuron Pedal 3 elicits foot margin lift and receives modulatory input from flow receptors. To assess the contribution of this single cell in turning behavior, two fine wires were glued to the surface of the brain over left and right Pedal 3. We determined that Pedal 3 activity is correlated with subsequent ipsilateral turns, preceding the lift of the foot margin and the change in orientation by a consistent interval. Both Pedal 3 cells show synchronous bursts of activity, and the firing frequency of the ipsilateral Pedal 3 increased before turns were observed to that side. Stimulation of the electrode over Pedal 3 proved sufficient to elicit an ipsilateral turn in Tritonia.     

The nature of sources of bioelectric and biomagnetic fields.

The goal of this paper is to examine the origins and relative importance of primary and secondary sources of electric and magnetic fields for excitable tissue. It is shown that for axonal and cardiac tissue a comparison of the relative field strength from both primary and secondary sources shows only the latter to be significant. Even if the divergence and curl of the primary source were independent (and hence were both needed to define the primary source), because the secondary sources all arise from the divergence of the primary source the magnetic field reflects the same source component as the electric field. As a consequence magnetic and electric fields arising from active tissue are strongly linked.     

The topological organization of primary afferents in the terminal ganglion of crayfish,Procambarus clarkii

Summary The central projections of primary afferents in the terminal ganglion of the crayfish can be seen when an axonal filling with nickel chloride with subsequent silver intensification was used for identification. We describe here the topological relationships of the projections to the landmark structures of the neuropil.The terminal ganglion has five pairs of sensory nerves associated with the mechanosensory hairs and internal proprioceptors. The projection fields of the primary sensory neurons in the nerves Rl and R2 are almost entirely restricted to the ipsilateral half of the ganglion, whereas those of the nerves R3, R4 and R5 cross the midline to form three sensory commissures, A6SCI, A7SCI and A7SCII. The projection fields are segregated from each other, although all are restricted to the ventral neuropil which lies under the ventral intermediate tract (VIT). The intersegmental projections that ascend via the connective ipsilateral to their origins could be observed. This pattern of projection correlates well with the receptive fields exhibited by several mechanosensory interneurons on the body surface of the final segment.     

Chronic excitotoxin-induced axon degeneration in a compartmented neuronal culture model

Glutamate excitotoxicity is a major pathogenic process implicated in many neurodegenerative conditions, including AD (Alzheimer''s disease) and following traumatic brain injury. Occurring predominantly from over-stimulation of ionotropic glutamate receptors located along dendrites, excitotoxic axonal degeneration may also occur in white matter tracts. Recent identification of axonal glutamate receptor subunits within axonal nanocomplexes raises the possibility of direct excitotoxic effects on axons. Individual neuronal responses to excitotoxicity are highly dependent on the complement of glutamate receptors expressed by the cell, and the localization of the functional receptors. To enable isolation of distal axons and targeted excitotoxicity, murine cortical neuron cultures were prepared in compartmented microfluidic devices, such that distal axons were isolated from neuronal cell bodies. Within the compartmented culture system, cortical neurons developed to relative maturity at 11 DIV (days in vitro) as demonstrated by the formation of dendritic spines and clustering of the presynaptic protein synaptophysin. The isolated distal axons retained growth cone structures in the absence of synaptic targets, and expressed glutamate receptor subunits. Glutamate treatment (100 μM) to the cell body chamber resulted in widespread degeneration within this chamber and degeneration of distal axons in the other chamber. Glutamate application to the distal axon chamber triggered a lesser degree of axonal degeneration without degenerative changes in the untreated somal chamber. These data indicate that in addition to current mechanisms of indirect axonal excitotoxicity, the distal axon may be a primary target for excitotoxicity in neurodegenerative conditions.