Convergent inputs to single neurons in two different subdivisions of somatosensory forepaw digit cortex of the raccoon.

The aim of this study was to compare the physiological properties of single neurons in the glabrous (G) and heterogeneous (H) subdivisions of primary somatosensory digit 3 cortex of adult raccoons. Extracellular recordings were obtained from 50 G neurons whose receptive fields (RFs) were confined to the glabrous skin of a digit, and 41 H neurons whose RFs were located on hairy skin, claws, or mixtures of skin types. Both electrical and mechanical stimulation of the digits were used to assess excitatory neuronal responsiveness. The two sets of neurons, which had nearly identical depth distributions, differed considerably in their input convergence: (i) the percentage of neurons (%N) responding to electrical or mechanical stimulation of each off-focus digit and (ii) the number of digits from which individual cells could be driven were significantly greater for H neurons. Those G and H cells which could be excited by off-focus inputs were examined for probability of response (P), number of spikes per response (S/R), and latency of response (L) to digit stimulation. Surprisingly, for input from any one digit, there were no significant differences in these response properties between the two sets of neurons. However, inputs from different (on-focus versus off-focus) digits varied significantly and revealed patterns of response properties that were qualitatively similar for both G and H neurons. Specifically, %N and P decreased while L increased symmetrically with distance of each off-focus digit from the central on-focus digit 3, reflecting corresponding variations in the synaptic accessibility and conduction time of off-focus excitatory inputs. In contrast, S/R values were very similar for all digits, suggesting that the synaptic strength of off-focus inputs is regulated independently of accessibility. Finally, preliminary findings indicated that denervation of the third digit caused a decrease in off-focus response latencies, while the normal latency profile across digits was retained. This suggests that the previously existing pattern of off-focus inputs to G and H neurons provides a template for denervation-induced cortical reorganization, whereby the synaptic efficacy of off-focus inputs is increased by disinhibition or facilitation.

Influence of grafting a smaller target muscle on the magnitude of naturally occurring trochlear motor neuron death during development.

About half of the motor neurons produced by some neural centers die during the course of normal development. It is thought that the size of the target muscle determines the number of surviving motor neurons. Previously, we tested the role of target size in limiting the number of survivors by forcing neurons to innervate a larger target (Sohal et al., '86). Results did not support the size-matching hypothesis because quail trochlear motor neurons innervating duck superior oblique muscle were not rescued. We have now performed the opposite experiment, i.e., forcing neurons to innervate a smaller target. By substituting the embryonic forebrain region of the duck with the same region of the quail before cell death begins, chimera embryos were produced that had a smaller quail superior oblique muscle successfully innervated by the trochlear motor neurons of the duck. The number of surviving trochlear motor neurons in chimeras was significantly higher than in the normal quail but less than in the normal duck. The smaller target resulted in some additional loss of neurons, suggesting that the target size may regulate neuron survival to a limited extent. Failure to achieve neuron loss corresponding to the reduction in target size suggests that there must be other factors that regulate neuron numbers during development.

Raccoon forelimb motorsensory cortex: I. Somatic afferent inputs to different cytoarchitectonic areas.

The distribution of potentials evoked in and around forelimb MsI by graded electrical stimulation of forelimb nerves has been studied in the raccoon (Procyon lotor). These data have been correlated with cytoarchitectonic characteristics of pericruciate cortical tissue. Potentials evoked by cutaneous nerve stimulation were widely distributed in MsI and SmI, but were smaller in amplitude and of longer latency in MsI than in SmI. Stimulation of ulnar, median or deep radial nerve at 1-1.4T, a strength considered to activate only Group I muscle afferent fibers, caused evoked potentials in a localized region mostly confined to posterior sigmoid gyrus. On the basis of cytoarchitectonic features it is concluded that: a) Anterior sigmoid gyrus, to near the level of the tip of the cruciate sulcus, is area 6 cortex; b) The lateral portion of the posterior sigmoid gyrus, cortex comprising the caudal bank of the cruciate sulcus and cortex surrounding the lateral tip of the cruciate sulcus is area 4 cortex; c) The middle portion of the posterior sigmoid gyrus, almost to the lip of the cruciate sulcus rostrally and extending onto the rostral bank of the ascending coronal and postcruciate sulci caudally, is area 3a cortex. The cortical focus for Group I afferent-evoked potentials coincides with area 3a cortex. It is concluded that forelimb MsI of raccoon is organized in a fashion similar to MsI of cats and monkeys.

Raccoon forelimb motorsensory cortex: II. Somatosensory inputs to single neurons.

Somatosensory input to 431 neurons in MsI has been studied in unanesthetized, paralyzed raccoons (Procyon lotor). The type of sensory input to neurons in lateral sigmoid gyrus (cytoarchitectonic area 4) and in posterior sigmoid gyrus (areas 4 and 3a) was not significantly different. Of these neurons, 36% were activated by superficial cutaneous stimulation (touch, tap or hair deflection) and 26% by deep stimulation (pressure or joint movement). Mute neurons (not driven by any form of peripheral stimulation tested, or vaguely driven) comprised 38% of the sample. Only 4% of anterior sigmoid gyrus (area 6) neurons responded to superficial or deep stimulation; 96% were mute. The majority of MsI neurons had small (less than or equal to 20 cm2) peripheral receptive fields (PRFs). There was a statistically significant trend for PRF size to decrease along the proximal-distal axis of the forelimb. The area of MsI digit PRFs was significantly larger than the area of SmI digit PRFs. Comparing the data for raccoon MsI with information from the literature for cats and monkeys suggests that the type and amount of somesthetic afferent input to forelimb MsI is related to the behavioral uses to which each animal puts the forelimb.

Motor outflow to cervical motoneurons from raccoon motorsensory cortex.

Motor outflow from forelimb motorsensory cortex (MsI) to forelimb muscle motoneurons in raccoon has been investigated using three approaches: 1) determination of latencies for cortically evoked efferent discharge in forelimb nerves; 2) determination of latencies for cortical facilitation of forelimb monosynaptic reflexes; and 3) intracellular recording of cortically evoked synaptic potentials. All three approaches indicated a major polysynaptic pathway (minimally disynaptic) for corticofugal facilitation or inhibition of cervical motoneurons. Suggestive evidence for a monosynaptic connection between forelimb MsI and cervical motoneurons was found for only one motoneurons. Nevertheless, the motor pathway between MsI and cervical motoneurons was shown to be more efficacious (defined on the basis of central delays) than in the cat under similar experimental conditions. The results are discussed in terms of organization changes in forelimb MsI which appear to be related to the extent to which certain mammals use their forelimbs for manipulating and exploring objects.

Limitations on impulse conduction at the branch point of afferent axons in frog dorsal root ganglion.

Impulse conduction at the branch point of afferent axons in dorsal root ganglion (DRG) has been studied using intracellular recording from frog DRG neurons in vitro. The least conduction interval (LCI, the minimum inter-response interval) was determined for pairs of impulses to successfully propagate through the branch point into the dorsal root. At 21 degrees-23 degrees C, average branch point LCI was significantly longer than for afferent fibers in the peripheral nerve. This result suggested that the branch point would limit the maximum frequency of action potentials that could conduct into the dorsal root. This was found to be the case. The maximum frequency of impulses in short trains (less than or equal to 40 ms) which could conduct into the dorsal root without failure (363 Hz) was accurately predicted by branch point LCI and was far less than the maximum frequency predicted from the LCI of axons in the peripheral nerve (610 Hz). Branch point LCI was correlated (r = -0.78) with the natural log of peripheral axon conduction velocity (CV). However, the relationship of LCI and CV was different for different types of neurons and the shape of the somatic action potential was found to be a reliable predictor of branch point LCI. Neurons with long-duration somatic action potentials with a shoulder on the falling phase tended to have low CV and invariably had long LCI's. Neurons with brief, smooth action potentials had short LCI's regardless of CV. These cells, which appear to be the most differentiated type, have found a way to minimize branch point LCI which is virtually independent of their axonal CV. For the latter neurons, branch point LCI was correlated (r = 0.42) with the reciprocal of the hyperpolarization level, at the cell body, required to block conduction through the branch point, suggesting that the proximity of the cell body to the branch point might play a role in determining the LCI of some neurons. Over a range of 12 degrees C to around 35 degrees C, branch point LCI was inversely related and maximum firing frequency directly related to temperature. At high temperatures (30 degrees-40 degrees C) conduction failure occurred at sites having particularly long LCI's. It is concluded that a) these axon branch points act as low-pass filters and set the maximum frequency of conducted impulses that can access the central nervous system; b) certain varieties of DRG neurons exhibit more branch point filtering action than others; and c) warming, within limits, reduces branch point filtering action.

Influence of reduced neuron pool on the magnitude of naturally occurring motor neuron death.

The present investigation was undertaken to examine the role of peripheral competition in survival of motor neurons during development. A loss of approximately half of the trochlear motor neurons in duck and quail occurs during the course of normal embryogenesis. The number of motor neurons in the nucleus of quail prior to the onset of cell death is identical to the final number of survivors in the nucleus of duck embryos (about 1,300 neurons). In the present study competition at the peripheral target was decreased by reducing the number of trochlear motor neurons initially projecting to their target muscle. This was accomplished by substituting the midbrain of duck embryos with the same neural tissue from quail embryos. Midbrain transplantation was performed before motor axon outgrowth and normal cell death begin. The development of the motor neurons and their sole target of innervation, the superior oblique muscle, was examined by using a variety of techniques. The source of the grafted motor neurons and of a reduction in the size of the motor neuron pool was confirmed from histological sections and cell counts. The grafted motor neurons projected their axons into the appropriate peripheral target, which was determined by the use of HRP tracing technique. Counts of muscle fibers, motor endplates, and acetylcholine receptors and measurement of total muscle protein indicated that the size of the superior oblique muscle in the chimera embryos was similar to that of the normal duck but significantly larger than the muscle in quail embryos. Electrophysiological observations indicated that the grafted trochlear motor neurons made functional connections with the superior oblique muscle. Counts of the trochlear motor neurons after the period of cell death indicated an average of 1,310 neurons in the nucleus of duck, 772 in quail, and 690 in the chimera embryos. The number of motor neurons in the chimera embryos is not significantly different from that in the normal quail. In other words, in spite of reduced peripheral competition trochlear motor neuron death of normal magnitude occurred. Lack of increased cell survival in our study suggests that trochlear motor neurons do not compete for survival at the peripheral target.

Unequal branch point filtering action in different types of dorsal root ganglion neurons of frogs.

The influence of the intraganglionic branch point on impulse conduction in single neurons of frog dorsal root ganglia (DRG) has been determined by measuring the least interval at which it will conduct two action potentials into the dorsal root. At 21-23 degrees C, branch points of myelinated fibers had long least conduction intervals and low safety factors for orthodromic impulse conduction compared to nodes of Ranvier in peripheral nerve. DRG neurons with broad somatic spikes with a shoulder on the falling phase and slowly conducting myelinated or non-myelinated axons had the longest least conduction intervals (lowest safety factors). DRG neurons with brief somatic spikes with little or no shoulder on the falling phase had short least conduction intervals (higher safety factor) regardless of their conduction velocity. The results indicate that certain DRG neurons have found a way to minimize branch point filtering action.

Mechanisms of accommodation in different types of frog neurons.

Responses of individual spinal ganglion neurons, sympathetic ganglion neurons, and motoneurons of frogs to linearly rising currents were investigated utilizing microelectrodes for intracellular stimulation and recording. Spinal ganglion neurons exhibited rapid accommodation to linearly rising currents. Minimal current gradients (MCG's) required to excite these neurons (average value, 106 rheobases/sec) were of the same order of magnitude as for some nerve fibers. Although sympathetic ganglion neurons exhibited responses to lower current gradients than spinal ganglion neurons, distinct MCG's (average value, 26 rheobases/sec) could always be established. MCG's could not be detected in most motoneurons, even with current gradients as low as 0.6 rheobase/sec. A few motoneurons exhibited distinct MCG's (average value, 11 rheobases/sec). The failure of spinal ganglion neurons to respond to anything other than rapidly rising currents appears to be due primarily to the development of severe delayed rectification. The inability of sympathetic ganglion neurons to respond to low current gradients appears to depend not only on delayed rectification but also on increases in depolarization threshold. When present in motoneurons, accommodation appears to result from the same mechanisms responsible for its appearance in sympathetic ganglion neurons.