John E. Swett

Sensory neurons of the rat sciatic nerve

Experiments have been undertaken in this laboratory over recent years to accurately determine the numbers and sizes of somatic neurons which contribute to the normal sciatic nerve, at mid-thigh levels, of the adult, albino rat. This article is concerned with the dorsal root ganglion (DRG) neuron population of the sciatic nerve whose cell bodies were identified through retrograde labeling of cut branches of the sciatic with horseradish peroxidase (HRP) and/or its wheat germ conjugate (WGA-HRP). It is essential to understand the neuronal composition of the normal rat sciatic nerve if the consequences of aging, nerve injury, and surgical repair to improve functional regeneration are to be properly evaluated. Neuron counts were determined from camera-lucida paper drawings of all labeled profiles in DRGs L3-L6 at 100 x magnification. The profiles, obtained by labeling individual branches of the sciatic nerve (sural, lateral sural, tibial, peroneal, medial, and lateral gastrocnemius/soleus nerves) were traced from 40-microns-thick, serial, frozen sections. The sizes of the perikarya, areas and diameters, were determined by tracing the perimeters of the drawn profiles on a digitizing tablet. The tablets output was inputted directly into a specially designed computer spreadsheet which contained a mathematical table for correcting the split-cell error inherent to the sectioning process. Afferents from any given branch of the sciatic normally occupied two to three adjacent ganglia. Sciatic DRG neurons were normally located in lumbar ganglia L3-L6. Nearly 98-99% of all sciatic DRG perikarya resided in the L4 and L5 DRGs. The L6 DRG, traditionally regarded as an important contributor to the rat sciatic, contained merely 0.4% of its afferent neurons while the L3 ganglion, frequently overlooked as a contributor, contained 1.2% of the mid-thigh sciatic afferents. The mean size of rat DRG neurons was about 29 microns (550-600 microns2). The corrected counts revealed that the normal sciatic nerve (at mid-thigh levels), in rats between 2 and 12 months of age, contained a mean, total DRG neuron population of about 10,500 neurons. This is probably an underestimate by 3-5% of the true number due to occasional unreliable labeling of some of the small DRG neurons. It is estimated that the normal, mean number of sciatic DRG neurons of young to middle-aged rats lies somewhere between 10,500 and 11,000 +/- 2000. The data suggest that nearly 20% of all DRG neurons in the sciatic nerve supply muscle afferents. The vast majority of the remaining neurons are involved with innervation of the skin.(ABSTRACT TRUNCATED AT 400 WORDS)

The cutaneous contribution to the hamstring flexor reflex in the rat: an electrophysiological and anatomical study

The location and properties of the cutaneous receptive fields responsible for detecting the flexor withdrawal reflex in the posterior head of biceps femoris (pBF) and semitendinosus (ST) components of the hamstring muscle have been examined in unanaesthetized decerebrate rats, spinalized at T10-T11. Single alpha-motoneurone efferents were recorded from the nerve to pBF and the principal head of ST and their responses to ipsi- and contralateral hindlimb skin stimulation investigated. The efferents to both muscles characteristically had a low or absent background discharge and they all had mechanoreceptive fields on the ipsilateral foot. The mechanical threshold of these fields was high with no response to light touch or brush. Fifty-four percent of these units also had a smaller and weaker contralateral mechanoreceptive field. The only apparent difference between ST and pBF efferents was that more ST efferents had contralateral fields than pBF units. Noxious, hot and cold thermal stimuli applied to the ipsilateral foot activated 56% of the efferents. Mustard oil, a chemical irritant, produced a long-lasting flexor response when applied to the ipsilateral foot. The responses of these efferents to stimulation of A beta, A delta and C cutaneous afferents in the sural nerve were also studied. Short latency reflexes were elicited in all efferents by A beta inputs, longer latency reflexes were elicited in 64% by A delta inputs and very long latency responses with long afterdischarges were found in 73% of the units to C inputs. Retrograde labelling of the hamstring motoneurones with WGA-HRP indicated that they lay in ventrolateral lamina IX extending from the caudal portion of the third lumbar segment to the junction of the 5th and 6th lumbar segments. Transganglionic labelling of small diameter primary afferent terminals in the dorsal horn of cutaneous nerves innervating the foot revealed that the longitudinal distribution corresponded closely with that of the hamstring motor nucleus. The flex-or reflex in the spinal rat provides a useful model therefore, for studying how the input in nociceptive afferents is processed and transformed within the spinal cord, to produce appropriate outputs.

Motor versus sensory neuron regeneration through collagen tubules

&NA; Differences in regeneration of sensory and motor nerves were studied in rats to determine the effects of entubulation with collagen conduits. The rat sciatic nerve was repaired either with a 10‐mm saline‐filled gap or with a no‐gap end‐to‐end repair cuffed within collagen tubules. These repairs were compared with the standard epineurial repairs. The populations of regenerated motor and sensory neurons in the peroneal nerves of all repairs were compared against the populations of normal peroneal neurons using horseradish peroxidase retrograde labeling. The epineurial repair resulted in regeneration of 65 percent (409 ± 150) of motor neurons and 79 percent (2127 ± 516) of sensory neurons (n = 6). The no‐gap end‐to‐end repair in a collagen tubule resulted in regeneration of 53 percent (338 ± 203) of motor and 70 percent (1893 ± 794) of sensory neurons (n = 7). In the 10‐mm gap repair, only 6.2 percent (39 ± 18) of motor neurons but 63 percent (1710 ± 557) of sensory neurons regenerated (n = 5). These results show that collagen entubulation supports nerve regeneration in end‐to‐end nerve repairs comparably to standard epineurial suture repairs. With the 10‐mm gap repairs in collagen tubules, sensory neurons regenerated consistently better than motor neurons in the same environment. Therefore, intrinsic differences exist between motor and sensory neuron regeneration in the same nerve. (Plast. Reconstr. Surg. 102: 430, 1998.)

Short latency activation of pyramidal tract cells by Group I afferent volleys in the cat

1. The contralateral bulbar pyramids were explored with low impedance micro‐electrodes in cats anaesthetized with chloralose to reveal the effect of Group I afferent volleys (deep radial nerve of the forelimb) on pyramidal tract (Pt) cells.

Most Dorsal Root Ganglion Neurons of the Adult Rat Survive Nerve Crush Injury

Severe crush of the rat sciatic nerve does not result in any significant cell death among motor neurons (Swett et al., 1991a). The present study reports on the survival of the dorsal root ganglion (DRG) neurons in the same experiments. From 15 to 187 days after crush of the left sciatic nerve, the common peroneal or sural nerve was cut and labeled distal to the injury with a mixture of horseradish peroxidase (HRP) and its wheatgerm agglutinin conjugate (WGA:HRP). In other cases, the crush injury was made far enough distally on a peroneal or sural branch to permit labeling several millimeters proximal to the injury. The procedures for reconstructing the regenerated DRG neuron populations were identical to those used in an earlier study describing the normal sciatic DRG neuron populations in the rat (Swett et al., 1991b). The normal peroneal nerve contains 2699 +/- 557 DRG neurons. When the peroneal nerve was crushed near its point of origin from the sciatic and labeled 10 mm distal to the injury, 2186 +/- 163 DRG neurons were counted, suggesting a decrease of about 19% (p < 0.01). However, when the entire sciatic nerve was crushed, distal labeling of the peroneal nerve revealed a mean number of 2578 +/- 291 DRG neurons, an insignificant reduction (p > 0.2). When the peroneal nerve was labeled proximal to a peroneal crush site, a similar number of DRG neurons (2563 +/- 412) was counted. Results following sural nerve crush were similar. The sural nerve normally contains 1675 +/- 316 DRG neurons. When the nerve was labeled distal to the injury, 1558 +/- 64 DRG neurons were counted--a number almost identical to that found (1529 +/- 240) when this nerve was labeled proximal to the injury. The results demonstrate that within 6 months of severe crush injury of the rat sciatic nerve, the vast majority of DRG neurons survive and regenerate new axons distally beyond the injury site, presumably to reinnervate their original targets.

Analgesia with peripheral nerve stimulation: Absence of a peripheral mechanism

Abstract Electrical stimulation of peripheral nerve can bring pain relief to patients suffering from intractable pain of peripheral origin. Some investigators believe that the mechanism responsible for this effect is located in the central nervous system (CNS) while others suggest that it is a peripheral one involving a conduction block in small diameter afferent fibers, thereby preventing the arrival of nociceptive information to the CNS. To determine which of these mechanisms was more likely to be responsible for producing this effect we studied 9 patient volunteers who sought relief from severe chronic, post‐traumatic pain in their extremities by stimulation of electrode cuffs implanted on their peripheral nerves. Using normalized stimulation procedures on human nerve and reapplying them to peripheral nerve of the cat, we were able to reconstruct, to a degree of accuracy not previously described in human subjects, the functional events occurring in nerve at the site of stimulation when patients reported pain relief. This effect was reliably induced in 7 of our 9 subjects with a mean relative stimulus intensity of 1.3T, a value insufficient to activate A&dgr; or C fibers. No conduction failure appeared in A fibers of the cat with prolonged stimulation at 75 Hz at intensities 4 times greater than those causing pain relief in man. Under conditions governed by patient choice there was no evidence that nerve block accounted for the effect. The mechanism appears to depend upon normal conduction of action potentials in large diameter fibers.

Detection thresholds to stimulation of ventrobasal complex in cats

Cats were trained to indicate, by bar pressing for food rewards, their detection of stimulation of the ventrobasal (VB) complex delivered through implanted bipolar electrodes. By varying stimulus intensity it was possible to determine thresholds for detection. Scaling stimulus intensity relative to the appearance of a minimal evoked potential allowed comparisons between animals and also comparisons with results obtained by stimulation of peripheral nerve. Animals could detect VB stimulation, but only at stimulus intensities consistently stronger than those required for minimal appearance of an evoked response in ipsilateral primary somatosensory cortex. Results of VB activation differed from cutaneous nerve effects in that VB detection thresholds were markedly influenced by stimulus frequency. They were lowest at frequencies above 30 Hz and increased greatly at lower frequencies. Discomfort or pain did not seem to result even from relatively high stimulus intensities. The results compare well with observations obtained from stimulation of VB in humans. The appearance of an evoked cortical response is not necessarily correlated with behavior. Under appropriate conditions, behavior can be elicited predictably with minimal electrocortical activity; under other conditions detection may be absent even when large numbers of cortical neurons are activated. We suggest that regions of the cerebral cortex receiving thalamocortical projections from VB may not be essential in the detection process.