Evidence of left-right asymmetry in the spinal cord has been accumulating for some time. This report adds an anatomical example. For thoracic segments in the cat, the dorsal root entries of a given segment, at least for T5 to T9, were shown to have a tendency to be located more rostrally on the right than on the left. Measurements of the root positions have been brought together from notes in experimental protocols over nine years (36 animals). The rostral-most root of a segment on the right was found to lie up to 4.5 mm more rostrally than that on the left (mean 0.93 ± 1.05 mm, n = 86), while the caudal-most root was similarly asymmetric by up to 5.5 mm (mean 0.98 ± 1.72 mm, n = 30). These results are supported by very similar measurements made from the undamaged regions of spinal cords in a further 36 animals that had spinal cord lesions. Implications of the asymmetry are discussed.

Previous evidence from electrophysiological experiments in anaesthetized cats with a chronic lateral lesion of the lower thoracic spinal cord indicated an expansion of the functional projections of expiratory bulbospinal neurones (EBSNs) in the segment above the lesion, measured at 16 weeks post-lesion. Here we investigate connections made by the same EBSNs to motoneurones in that segment, using cross-correlations between their discharges. The connections to the internal intercostal nerve motoneurones were found to be no different from controls. However, a significant increase was found in the number of connections between EBSNs and γ motoneurones of the external intercostal nerve (8/24, compared to 1/16) with possibly additional connections to the α motoneurones of the same nerve. Increased connections to the γ motoneurones of the internal intercostal nerve could not be ruled out. The expanded functional projections are thus likely to include new connections to γ motoneurones. We suggest that γ motoneurones may be inherently more receptive to new inputs. If so, the previously discounted role of abnormal fusimotor discharges in motor disorders would be worth reconsideration.

The morphology and projections of ventral horn interneurones in the segment above an ipsilateral thoracic lateral spinal cord lesion were studied in the cat by intracellular injections of Neurobiotin at 6 to 18 weeks post-lesion and compared with previously published control data from uninjured spinal cords. The cell axons ascended, descended or both, mostly contralaterally and mostly spared by the lesion. Unusual morphological dendritic features were seen in the lesion group, mostly growth-related, including complex dendritic appendages, twisted or multiple-branched terminal dendrites, commissural dendrites, apparently swollen proximal dendrites and rostrocaudal asymmetries. Significant quantitative differences included more dendritic spines in the lesion group (3.4×) and smaller soma areas in the lesion group (with similar numbers of primary dendrites and rostrocaudal dendritic spans). Immunoreactivity to microtubule associated protein 2a/b was detected in the proximal, but not distal, dendrites of cells in the lesion group, corresponding to an overall decrease in immunoreactivity in the ventral horns on the lesion side compared to the other. For axon collaterals, significant increases for the lesion group were seen in the number of collaterals in the first 4 mm of axon and in the area of ventral/intermediate horn occupied by terminals, including increased innervation of some regions, among which were the intermediolateral columns. This dendritic and axonal plasticity makes the interneuones candidates for a role in detour circuits but also for a maladaptive role in autonomic hyperreflexia.

The contribution of branched-axon monosynaptic inputs in the generation of short-term synchronization of motoneurones remains uncertain. Here, synchronization was measured for intercostal and abdominal motoneurones supplying the lower thorax and upper abdomen, mostly showing expiratory discharges. Synchronization in the anaesthetized cat, where the motoneurones receive a strong direct descending drive, is compared with that in anaesthetized or decerebrate rats, where the direct descending drive is much weaker. In the cat, some examples could be explained by branched-axon monosynaptic inputs, but many others could not, by virtue of peaks in cross-correlation histograms whose widths (relatively wide) and timing indicated common inputs with more complex linkages, e.g., disynaptic excitatory. In contrast, in the rat, correlations for pairs of internal intercostal nerves were dominated by very narrow peaks, indicative of branched-axon monosynaptic inputs. However, the presence of activity in both inspiration and expiration in many of the nerves allowed additional synchronization measurements between internal and external intercostal nerves. Time courses of synchronization for these often consisted of combinations of peaks and troughs, which have never been previously described for motoneurone synchronization and which we interpret as indicating combinations of inputs, excitation of one group of motoneurones being common with either excitation or inhibition of the other. Significant species differences in the circuits controlling the motoneurones are indicated, but in both cases, the roles of spinal interneurones are emphasised. The results demonstrate the potential of motoneurone synchronization for investigating inhibition and have important general implications for the interpretation of neural connectivity measurements by cross-correlation.

Introduction : HF-SCS applied to the ventral surface of the upper thoracic spinal cord (T2 level) results in physiologic activation of both the diaphragm and inspiratory intercostal muscles. Objective: To evaluate the effects of HF-SCS (500Hz, 1mA) applied to the ventral surface of the entire thoracic spinal cord. Material and Methods: In 4 anesthetized dogs, three eight plate stimulation leads, were inserted epidurally and positioned on the ventral surface of the spinal cord via laminectomies at the T6 and T10 levels. Changes in airway pressure (P) were assessed over different regions of the thoracic spinal cord following electrical stimulation (HFSCS: 500 Hz, 1 mA, 0.2ms) at each electrode lead in separate trials, under conditions of airway occlusion and hyperventilation induced apnea. Results: HF-SCS (500Hz, 1mA) resulted in negative P generation over the ventral surface of the entire thoracic spinal cord. Negative P generation was most pronounced at the T2 level (60±2 cmH2O) and T9 levels (46±8 cmH2O). Further examination of HFSCS at the T9 level demonstrated that maximum negative P was dependent upon low stimulus currents (1-3 mA) and high stimulus frequencies (>300 Hz). Both the diaphragm and inspiratory intercostal muscles contributed to negative P generation. Conclusions: The unique findings of the present investigation include the demonstration that spinal circuits which have connections to the inspiratory motoneuron pools exist over the ventral surface of the entire thoracic spinal cord. We postulate that responses are mediated, in part, by stimulation of the previously described intercostal to phrenic excitatory reflex. The large negative P generated at the T9 level suggests that HF-SCS in this region of the spinal cord may be a useful method to restore breathing in ventilator dependent tetraplegics.

In animals, high-frequency spinal cord stimulation (HF-SCS) applied on the ventral epidural surface at the T2 level results in negative airway pressure generation consistent with inspiratory muscle activation. In the present study, in anesthetized dogs, we found that ventral HF-SCS (500 Hz) applied at all thoracic levels resulted in negative airway pressure generation. In the region of the lower thoracic spinal cord, negative airway pressure generation was most pronounced at the T9 level. At this level, airway pressure generation was monitored: 1) during ventral HF-SCS over a wide range of stimulus amplitudes (0.5-15 mA) and frequencies (50-1,000 Hz) and 2) following spinal sections at C8 (to assess potential diaphragm activation) and subsequently at T6 (to assess potential intercostal muscle activation). The application of low stimulus currents between 1 and 2 mA and high stimulus frequencies (>300 Hz) resulted in the development of large negative airway pressure generation. Stimulation with 1 mA, 500 Hz resulted in a highest negative airway pressure generation of 47 ± 2 cmH2O. Increasing stimulus current was associated with progressive reductions in the magnitude of negative airway pressure generation. HF-SCS (500 Hz) with 15 mA resulted in a negative airway pressure generation of 7 ± 3 cmH2O. C8 section markedly reduced negative airway pressure generation, and subsequent T6 section resulted in positive airway pressure generation after HF-SCS. Our results indicate the existence of pathways with connections to both the phrenic and inspiratory intercostal motoneuron pools in the ventral part of the lower thoracic spinal cord. We speculate that the circuits mediating the previously described excitatory intercostal-to-phrenic reflex mediate the observed responses. NEW & NOTEWORTHY This study suggests that, in contrast to dorsal high-frequency spinal cord stimulation at the T9 spinal level, which results in positive pressure generation, ventral high-frequency spinal cord stimulation at the same spinal level results in large negative airway pressure generation with low stimulus currents. This method, therefore, may provide an alternative method to restore ventilation in ventilator-dependent spinal cord-injured patients.

This feature article focuses on the discrepancy between the distribution of axon diameters within the primate corticospinal tract, determined neuroanatomically, and the distribution of axonal conduction velocities within the same tract, determined electrophysiologically. We point out the importance of resolving this discrepancy for a complete understanding of corticospinal functions, and discuss the various explanations for the mismatch between anatomy and physiology.

There are hardly any published data on the characteristics of muscle nerve sympathetic discharges occurring in parallel with the somatic motoneurone discharges in the same nerves. Here, we take advantage of the naturally occurring respiratory activity in recordings of efferent discharges from branches of the intercostal and abdominal nerves in anesthetized cats to make this comparison. The occurrence of efferent spikes with amplitudes below that for alpha motoneurones were analyzed for cardiac modulation, using cross-correlation between the times of the R-wave of the ECG and the efferent spikes. The modulation was observed in nearly all recordings, and for all categories of nerves. It was strongest for the smallest amplitude spikes or spike-like waveforms, which were deduced to comprise postsynaptic sympathetic discharges. New observations were: (1) that the cardiac modulation of these discharges was modest compared to most previous reports for muscle nerves; (2) that the amplitudes of the sympathetic discharges compared to those of the somatic spikes were strongly positively correlated to nerve diameter, such that, for the larger nerves, their amplitudes overlapped considerably with those of gamma motoneurone spikes. This could be explained by random summation of high rates of unit sympathetic spikes. We suggest that under some experimental circumstances this overlap could lead to considerable ambiguity in the identity of the discharges in efferent neurograms.

Previous analyses of recordings of alpha motoneuron discharges from branches of the intercostal and abdominal nerves in anesthetized cats under neuromuscular blockade demonstrated modulation with the cardiac cycle. This modulation was interpreted as evidence that thoracic somatosensory afferents, most likely muscle spindles, provide a signal to the CNS that could contribute to cardiac interoception. Here, two aspects of these observations have been extended. First, new measurements of thoracic and abdominal EMG activity in spontaneously breathing dogs show that a very similar modulation exists in these rather different circumstances. Second, further analysis of the cat recordings shows that cardiac modulation of the discharges of bulbospinal neurons that transmit the expiratory drive to thoracic motoneurons is weak and of an inappropriate time-course to be a contributor to the effect seen in the motoneurons.

Recordings of alpha motoneuron discharges from branches of the intercostal and abdominal nerves in anesthetized cats were analyzed for modulation during the cardiac cycle. Cardiac modulation was assessed by the construction of cross-correlation histograms between the R-wave of the ECG and the largest amplitude efferent spikes. In all but two recordings (which were believed to have either no or few alpha spikes), the histograms showed relatively short duration peaks and/or troughs (widths at half amplitude 4-50 ms) at lags of 10-150 ms. These observations were deduced to result from activity in oligosynaptic pathways, probably from muscle spindle afferents, whose discharges are known to be synchronized to the cardiac pulse. The results suggest that onward transmission of the cardiac signal from thoracic muscle afferents (and possibly from other dynamically sensitive afferents) to other parts of the central nervous system is highly likely and that therefore these afferents could contribute to cardiac interoception. NEW & NOTEWORTHY It has been recognized since 1933 that muscle spindles respond to the cardiac pulse, but it is unknown whether this cardiac signal is transmitted to other levels in the nervous system. Here we show that a cardiac signal, likely arising from muscle spindles, is present in the efferent activities of thoracic and abdominal muscle nerves, suggesting probable onward transmission of this signal to higher levels and therefore that muscle spindles could contribute to cardiac interoception.

to the editor: In recent years, De Luca and colleagues ([De Luca et al. 1993][1]; [De Luca and Kline 2014][2]) have expressed considerable scepticism for the conventionally accepted view that motoneuron synchronization results from activity in common inputs to the motoneurons concerned. Now [Kline

A previous neurophysiological investigation demonstrated an increase in functional projections of expiratory bulbospinal neurons (EBSNs) in the segment above a chronic lateral thoracic spinal cord lesion that severed their axons. We have now investigated how this plasticity might be manifested in thoracic motoneurons by measuring their respiratory drive and the connections to them from individual EBSNs. In anesthetized cats, simultaneous recordings were made intracellularly from motoneurons in the segment above a left-side chronic (16 wk) lesion of the spinal cord in the rostral part of T8, T9, or T10 and extracellularly from EBSNs in the right caudal medulla, antidromically excited from just above the lesion but not from below. Spike-triggered averaging was used to measure the connections between pairs of EBSNs and motoneurons. Connections were found to have a very similar distribution to normal and were, if anything (nonsignificantly), weaker than normal, being present for 42/158 pairs, vs. 55/154 pairs in controls. The expiratory drive in expiratory motoneurons appeared stronger than in controls but again not significantly so. Thus we conclude that new connections made by the EBSNs following these lesions were made to neurons other than α-motoneurons. However, a previously unidentified form of functional plasticity was seen in that there was a significant increase in the excitation of motoneurons during postinspiration, being manifest either in increased incidence of expiratory decrementing respiratory drive potentials or in an increased amplitude of the postinspiratory depolarizing phase in inspiratory motoneurons. We suggest that this component arose from spinal cord interneurons.

In this study we investigated how the networks mediating respiratory and locomotor drives to lumbar motoneurons interact and how this interaction is modulated in relation to periodic variations in blood pressure (Mayer waves). Seven decerebrate cats, under neuromuscular blockade, were used to study central respiratory drive potentials (CRDPs, usually enhanced by added CO2) and spontaneously occurring locomotor drive potentials (LDPs) in hindlimb motoneurons, together with hindlimb and phrenic nerve discharges. In four of the cats both drives and their voltage-dependent amplification were absent or modest, but in the other three, one or other of these drives was common and the voltage-dependent amplification was frequently strong. Moreover, in these three cats the blood pressure showed marked periodic variation (Mayer waves), with a slow rate (periods 9-104 s, mean 39 ± 17 SD). Profound modulation, synchronized with the Mayer waves was seen in the occurrence and/or in the amplification of the CRDPs or LDPs. In one animal, where CRDPs were present in most cells and the amplification was strong, the CRDP consistently triggered sustained plateaux at one phase of the Mayer wave cycle. In the other two animals, LDPs were common, and the occurrence of the locomotor drive was gated by the Mayer wave cycle, sometimes in alternation with the respiratory drive. Other interactions between the two drives involved respiration providing leading events, including co-activation of flexors and extensors during post-inspiration or a locomotor drive gated or sometimes entrained by respiration. We conclude that the respiratory drive in hindlimb motoneurons is transmitted via elements of the locomotor central pattern generator. The rapid modulation related to Mayer waves suggests the existence of a more direct and specific descending modulatory control than has previously been demonstrated.

Internal intercostal and abdominal motoneurons are strongly coactivated during expiration. We investigated whether that synergy was paralleled by synergistic Group I reflex excitation. Intracellular recordings were made from motoneurons of the internal intercostal nerve of T8 in anesthetized cats, and the specificity of the monosynaptic connections from afferents in each of the two main branches of this nerve was investigated. Motoneurons were shown by antidromic excitation to innervate three muscle groups: external abdominal oblique [EO; innervated by the lateral branch (Lat)], the region of the internal intercostal muscle proximal to the branch point (IIm), and muscles innervated from the distal remainder (Dist). Strong specificity was observed, only 2 of 54 motoneurons showing excitatory postsynaptic potentials (EPSPs) from both Lat and Dist. No EO motoneurons showed an EPSP from Dist, and no IIm motoneurons showed one from Lat. Expiratory Dist motoneurons fell into two groups. Those with Dist EPSPs and none from Lat (group A) were assumed to innervate distal internal intercostal muscle. Those with Lat EPSPs (group B) were assumed to innervate abdominal muscle (transversus abdominis or rectus abdominis). Inspiratory Dist motoneurons (assumed to innervate interchondral muscle) showed Dist EPSPs. Stimulation of dorsal ramus nerves gave EPSPs in 12 instances, 9 being in group B Dist motoneurons. The complete absence of heteronymous monosynaptic Group I reflex excitation between muscles that are synergistically activated in expiration leads us to conclude that such connections from muscle spindle afferents of the thoracic nerves have little role in controlling expiratory movements but, where present, support other motor acts.

Small axons far outnumber larger fibers in the corticospinal tract, but the function of these small axons remains poorly understood. This is because they are difficult to identify, and therefore their physiology remains obscure. To assess the extent of the mismatch between anatomic and physiological measures, we compared conduction time and velocity in a large number of macaque corticospinal neurons with the distribution of axon diameters at the level of the medullary pyramid, using both light and electron microscopy. At the electron microscopic level, a total of 4,172 axons were sampled from 2 adult male macaque monkeys. We confirmed that there were virtually no unmyelinated fibers in the pyramidal tract. About 14% of pyramidal tract axons had a diameter smaller than 0.50 μm (including myelin sheath), most of these remaining undetected using light microscopy, and 52% were smaller than 1 μm. In the electrophysiological study, we determined the distribution of antidromic latencies of pyramidal tract neurons, recorded in primary motor cortex, ventral premotor cortex, and supplementary motor area and identified by pyramidal tract stimulation (799 pyramidal tract neurons, 7 adult awake macaques) or orthodromically from corticospinal axons recorded at the mid-cervical spinal level (192 axons, 5 adult anesthetized macaques). The distribution of antidromic and orthodromic latencies of corticospinal neurons was strongly biased toward those with large, fast-conducting axons. Axons smaller than 3 μm and with a conduction velocity below 18 m/s were grossly underrepresented in our electrophysiological recordings, and those below 1 μm (6 m/s) were probably not represented at all. The identity, location, and function of the majority of corticospinal neurons with small, slowly conducting axons remains unknown.

Cross-correlation of neural discharges was used to investigate the connections between expiratory bulbospinal neurons (EBSNs) in the caudal medulla and expiratory motoneurons innervating thoracic and abdominal muscles in anesthetized cats. Peaks were seen in the cross-correlation histograms for around half of the EBSN-nerve pairs for the following: at T8, the nerve branches innervating internal intercostal muscle and external abdominal oblique muscle and a more distal branch of the internal intercostal nerve; and at L1, a nerve branch innervating internal abdominal oblique muscle and a more distal branch of the ventral ramus. Fewer peaks were seen for the L1 nerve innervating external abdominal oblique, but a paucity of presumed α-motoneuron discharges could explain the rarity of the peaks in this instance. Taking into account individual EBSN conduction times to T8 and to L1, as well as peripheral conduction times, nearly all of the peaks were interpreted as representing monosynaptic connections. Individual EBSNs showed connections at both T8 and L1, but without any discernible pattern. The overall strength of the monosynaptic connection from EBSNs at L1 was found to be very similar to that at T8, which was previously argued to be substantial and responsible for the temporal patterns of expiratory motoneuron discharges. However, we argue that other inputs are required to create the stereotyped spatial patterns of discharges in the thoracic and abdominal musculature.