Dorsal Horn Plasticity Following Re-routeing of Peripheral Nerves: Evidence for Tissue-Specific Neurotrophic Influences from the Periphery

Some properties of primary sensory neurons change when they reinnervate new peripheral targets (McMahon et al., Neuroscience, 33, 67 - 75, 1989). We ask here if such influences can extend to the central connectivity of sensory neurons. In adult rats the nerve to the gastrocnemius muscle (GN) and the cutaneous sural nerve (SN) were self- and cross-anastomosed on left- and right-hand sides, respectively, so that they regenerated to either appropriate or inappropriate targets. Ten to 14 weeks later, the distribution and strength of spinal connections of the SN and GN were determined. The unmyelinated afferents in the GN innervating skin increased their connectivity to 286% of that seen for the GN innervating muscle (P < 0.005), and came to resemble normal cutaneous afferents. However, for the SN there was no significant difference between appropriately and inappropriately regenerated nerves by this measure. The ability of myelinated fibres to produce inhibitions and facilitations in dorsal horn cells was also assessed. The intact or self-anastomosed SN produced predominantly inhibitory effects, whilst the GN produced predominantly facilitatory effects. After the SN had regenerated to muscle its central effects became predominantly facilitatory, whilst those of the GN innervating skin became inhibitory. These changes were statistically significant. In conclusion, we have found that major changes in the physiology of central connections in the dorsal horn may occur following peripheral reinnervation of foreign targets. The changes that were seen were appropriate to the new target, and could not easily be explained by non-specific changes due to axotomy, or changes in A-fibre-mediated inhibitions. We suggest that these effects might arise because of trophic influences arising in and specific to different peripheral targets.     

Muscle Afferents Innervating Skin Form Somatotopically Appropriate Connections in the Adult Rat Dorsal Horn

We have studied the somatotopic reorganization in dorsal horn neurons after a disruption in the normal spatial arrangement of primary sensory axons in adult rats. Muscle afferents were redirected to skin by cutting and cross-anastomosing the hindlimb gastrocnemius nerve (GN) and sural nerve (SN). It has previously been shown that after 10 – 12 weeks GN afferents innervate the hairy skin of the lateral ankle and calf (previously innervated by SN afferents) and become potentially capable of relaying information on the location and intensity of stimuli applied to the skin. We determined the receptive field and response properties of dorsal horn neurons in the lumbar spinal cord, in regions where the lower hindlimb is normally represented. In control animals (with intact or self-anastomosed sural nerves) very few neurons (<8%) received any synaptic input from the GN as assessed by electrical stimulation of the nerve. In contrast, when this nerve innervated skin, many cells responded to GN stimulation, and these nearly all had receptive field components in the former SN territory. Moreover, in animals with cross-anastomosed nerves, cells without GN inputs all had receptive fields outside the former SN skin territory. We have shown that in all likelihood GN afferents substituted for SN afferents in subserving the low and high threshold receptive fields of dorsal horn neurons. Furthermore, for many neurons, receptive fields were formed from inappropriately regrown GN afferents and adjacent intact cutaneous afferents (in the tibial or common peroneal nerves). Therefore, when GN afferents innervate skin in adult animals, they alter their central connectivity in an appropriate manner for their new peripheral terminations, so that an orderly somatotopic representation of the hind limb skin is maintained. We suggest that this plasticity of dorsal horn somatotopy is driven in part by activity-dependent mechanisms.     

The somatotopic organization of primary afferent terminals in the superficial laminae of the dorsal horn of the rat spinal cord

Transganglionic transport of wheatgerm agglutinin conjugated horse-radish peroxidase (WGA-HRP) was used to reveal the central distribution of terminals of primary afferent fibers from peripheral nerves innervating the hind leg of the rat. In separate experiments the sizes and locations of cutaneous peripheral receptive fields were determined by electrophysiological recording techniques for each of the nerves that had been labeled with WGA-HRP. By using digital image analysis, the sizes and positions of the peripheral receptive fields were correlated with the areas of superficial dorsal horn occupied by terminals of primary afferents from each of these receptive fields. Data were obtained from the posterior cutaneous nerve of the thigh, lateral sural, sural, saphenous, superficial peroneal, and tibial nerves. The subdivisions of the sciatic nerve, the sural, lateral sural, superficial peroneal, and tibial nerves each projected to a separate and distinct region of the superficial dorsal horn and collectively formed a "U"-shaped zone of terminal labeling extending from lumbar spinal segments L2 to the caudal portions of L5. The gap in the "U" extended from L2 to the L3-4 boundary and was occupied by terminals from the saphenous nerve. Collectively, all primary afferents supplying the hindlimb occupied the medial 3/4 of the superficial dorsal horn with terminals from the tibial nerve lying most medially and occupying the largest of all the terminal fields. Afferents from the superficial peroneal lay in a zone between the medially situated tibial zone and the more laterally placed sural zone. Afferents from the posterior cutaneous nerve were located most caudally and laterally. Terminal fields from the posterior cutaneous and saphenous nerves differed from the others in having split representations caused presumably by their proximity to the mid-axial line of the limb. Comparisons between the peripheral and the central representations of each nerve revealed that 1 mm2 of surface area of the superficial dorsal horn serves approximately 600-900 mm2 of hairy skin and roughly 300 mm2 of glabrous skin. The vast majority of terminal labeling observed in the dorsal horn was found in the marginal layer and substantia gelatinosa, suggesting that small diameter afferents have an orderly somatotopic arrangement in which each portion of the skin surface is innervated by afferent fibers that terminate in preferred localities within the dorsal horn.     

Mapping and plasticity of acid phosphatase afferents in the rat dorsal horn

After peripheral nerve injury in rats, naturally occuring fluoride resistant acid phosphatase (FRAP) disappears from the substantia gelatinosa in that part of the dorsal horn in which the injured nerve afferents terminate. We have taken advantage of this fact to establish the spinal distribution of nerves innervating the skin of the hindlimb. The spinal map of the foot, the distal part of the lower lumbar dermatomes, is in the medial part of the substantia gelatinosa. More proximal skin of the thigh and the lower back maps laterally. The zone of disappearance of FRAP after sciatic nerve section did not shrink detectably within the first few months after injury, provided that regeneration of the nerve was prevented. After one year, however, central FRAP activity was at least partially restored. Secondary transection of the sciatic nerve eliminated the new enzyme and transection of neighboring nerves failed to do so. The restored FRAP activity therefore reflects renewed synthesis and transport of enzyme in still injured neurons, and not central sprouting of intact neighboring afferents.     

Preferential Growth of Neonatal Rat Dorsal Root Ganglion Cells on Homotypic Peripheral Nerve Substrates In Vitro

Developing sensory neurons interact with molecular signals in the local environment to generate stereotypic nerve pathways. Regenerating neurons seem to lose the ability to reinnervate their original sites in the targets, resulting in abnormal sensory input and consequent clinical pathophysiology. The specificity of reinnervation of peripheral targets by regenerating axons is thus crucial for normal recovery of function. In this study, we have examined evidence for selectivity of interactions between primary afferent neurons from identified levels of the spinal cord and different peripheral nerve environments by culturing these neurons on sections of nerves to muscle and viscera. We have compared the growth of a population of sensory afferents normally innervating somatic targets (dorsal root ganglion cells from L4 and L5) with populations containing many afferents innervating visceral targets (L6 and S1 dorsal root ganglia and nodose ganglion). These neurons, from newly born rats, were cultured on unfixed cryostat sections of normal and prelesioned gastrocnemius nerve, pelvic spinal nerve and vagus nerve from adult rats. Normal muscle nerve was seen to support the regeneration of a significantly greater proportion of somatic neurons, with longer neurites, than the visceral nerves. Similarly, much higher proportions of the‘visceral’population of afferent neurons were seen to extend neurites on the normal visceral nerve substrates, with longer neurites, than on the muscle nerve substrate. The selectivity displayed by the sensory neurons for their normal nerve substrates was abolished when they were cultured on prelesioned nerve substrates subjected to Wallerian degeneration, which was apparent from the equivalent and increased proportions of growing neurons having comparable neurite lengths, on all the nerve substrates. We conclude that sensory neurons recognize and respond to substrate-specific and substrate-bound molecules present in normal adult peripheral nerves, and that these differences are lost in prelesioned nerves following Wallerian degeneration.     

Morphology and somatotopic organization of the central terminals of hindlimb hair follicle afferents in the rat lumbar spinal cord

The morphology of the central collateral arborizations of 24 A-beta hair follicle afferents (HFAs) innervating different regions of the skin of the hindlimb were studied by the intra-axonal injection of horseradish peroxidase (HRP) in adult rats. A total of 236 collaterals were recovered. These fell into three classes--complex, simple, and blind-ending--based on numbers of boutons and terminal branch patterns. The morphology of the HFA central arbors innervating the lateral and medial leg and dorsum of the foot was flame-shaped. Afferents with receptive fields on the glabrous-hairy skin border consistently had extra terminal branches running ventromedially into laminae IV/V. Differences in the width of terminal arbors were found. HFA terminals innervating the lateral leg formed narrower sheets than those innervating the dorsum of the foot and toes. The somatotopic organization of the collaterals and terminal arborizations of individual afferents were analyzed both by considering all the collaterals along an axon's rostrocaudal extent and by only examining arbors with boutons (the complex and simple arbors). Thirty-seven percent of blind-ending and 18% of simple collaterals were found to overlap in the rostrocaudal direction with the complex arborizations of afferents whose receptive fields were in a different cutaneous nerve territory. There was no overlap between complex arborizations of afferents from different nerve territories. However, the complex arbors of afferents with receptive fields within a particular nerve territory showed considerable terminal overlap even if they had nonadjacent peripheral receptive fields. The topographical organization of the central terminals of HFAs, forms a coarse somatotopic map of overlapping terminals whereby a particular region of dorsal horn has a maximal, but not exclusive, input from a particular area of skin.     

Effect of peripheral nerve injury on receptive fields of cells in the cat spinal cord

When the sciatic and saphenous nerves are cut and ligated in adult cats, the immediate effect is the production of a completely anesthetic foot and a region in medial lumbar dorsal horn where almost all cells have lost their natural receptive fields (RFs). Beginning at about 1 week and maturing by 4 weeks, some 40% of cells in the medial dorsal horn gain a novel RF on proximal skin, that is, upper and lower leg, thigh, lower back, or perineum. This new RF is supplied by intact proximal nerves and not by sciatic and saphenous nerve fibers that sprouted in the periphery. During the period of switching of RFs from distal to proximal skin there was no gross atrophy of dorsal horn grey matter and no Fink-Heimer stainable degeneration of central arbors and terminals of peripherally axotomized afferents. In intact animals medial dorsal horn cells showed no sing of response to mechanical stimulation of proximal skin. RFs of some of the cells had spontaneous variations in size and sensitivity, but these were not nearly sufficient to explain the large shifts observed after chronic nerve section. Tetanic electrical stimulation of skin or peripheral nerves often caused RFs to shrink, but never to expand. Although natural stimuli of proximal skin would not excite medial dorsal horn cells in intact or acutely deafferented animals, it was found that electrical stimulation of proximal nerves did excite many of these cells, often at short latencies. In the discussion we justify our working hypothesis that the appearance of novel RFs is due to the strengthening or unmasking of normally present but ineffective afferent terminals, rather than to long-distance sprouting of new afferent arbors within the spinal cord.     

Do central terminals of intact myelinated primary afferents sprout into the superficial dorsal horn of rat spinal cord after injury to a neighboring peripheral nerve?

In order to investigate whether normal myelinated primary afferent axons sprout into the territories of adjacent injured peripheral nerve fibers in the superficial dorsal horn of the spinal cord, adult rats underwent either sectioning of the saphenous or femoral nerves on one side, or else unilateral denervation of the skin of the posterior thigh. Two weeks later cholera toxin B subunit (CTb), which is normally transported selectively by myelinated somatic primary afferents, was injected into the ipsilateral (intact) sciatic nerve. The relationship between CTb, vasoactive intestinal peptide (VIP), and binding of Bandeiraea simplicifolia isolectin B4 (IB4) was then examined in the ipsilateral dorsal horn of the second to fifth lumbar spinal segments (L2-L5). Sectioning of the femoral or saphenous nerves resulted in a reduction of IB4 binding in laminae I-II in the medial third of the dorsal horn of L2, L3, and the upper part of L4. VIP-immunoreactivity was upregulated in exactly the same regions in which IB4-binding was reduced. These correspond to the areas that were previously innervated by unmyelinated afferents in the sectioned nerves. CTb-labeling was detected in regions known to receive input from myelinated sciatic afferents: lamina I and a band extending from the inner part of lamina II (IIi) to lamina V in the L3-5 segments, and the deepest part of the dorsal horn in L2. Importantly, no CTb-labeling was detected in the outer part of lamina II (IIo) in the denervated areas. Sectioning of branches of the posterior cutaneous nerve of the thigh resulted in a reduction of IB4-binding and upregulation of VIP-immunoreactivity in the lateral part of the superficial dorsal horn of caudal L4 and L5. Again, CTb-immunoreactivity showed the normal sciatic pattern in L4-L5, with no labeling detected in lamina IIo in the denervated region. These results do not support the suggestion that the central terminals of intact myelinated afferents sprout into regions of lamina II occupied by adjacent nerves that have been axotomized peripherally.     

Deafferentation-induced expansion of saphenous terminal field labelling in the adult rat dorsal horn following pronase injection of the sciatic nerve

We have examined the effect of the degeneration of sciatic nerve afferents on the distribution of saphenous terminals in the adult rat dorsal horn. Deafferentation was produced by injection into the sciatic nerve of pronase, a combination of proteolytic enzymes, which causes death of ganglion cells and degeneration of their terminal fields. The saphenous terminal fields were labelled by exposing the cut nerve to a combination of horseradish peroxidase (HRP) and wheat germ agglutinin-horseradish peroxidase (WGA-HRP). Terminals were mainly found in the superficial dorsal horn, indicating that small-diameter afferents were heavily labelled. In one group of control animals, the normal sciatic and normal saphenous terminal fields were shown to be bilaterally symmetrical. In the experimental group, the initial injection of one sciatic nerve with pronase was followed 4 months later by bilateral HRP/WGA-HRP labelling of both saphenous nerves. In each animal, the terminal field of the saphenous nerve on the lesioned side was expanded in the medial, lateral, and caudal directions. Medially and laterally, the expanded terminal field overlapped more of the sciatic territory than normal; caudally, saphenous terminals were found in the rostral portion of the L5 segment, in an area normally filled by sciatic terminals and devoid of saphenous terminals. The expansion resulted in a total saphenous area 26% larger than the control side. Electron microscopy demonstrated that the label in both the normal and expanded territories was primarily contained in axons and terminals, with minor transneuronal labelling. Labelled terminals in the expanded areas were both simple terminals with round, clear vesicles, and glomerular terminals with multiple synaptic contacts; these terminal types resemble those previously described for primary afferents in the superficial dorsal horn. Although the preexistence of "silent" synaptic terminals in the expanded areas cannot be disproven, the data support the hypothesis that primary afferents in the adult have the potential to sprout and establish synapses when the conditions of the deafferentation are favorable.     

Spinal cord projections from hindlimb muscle nerves in the rat studied by transganglionic transport of horseradish peroxidase, wheat germ agglutinin conjugated horseradish peroxidase, or horseradish peroxidase with dimethylsulfoxide

The spinal cord projections of four different groups of hindlimb muscle nerve branches--the medial and lateral gastrocnemius nerves, muscle branches of the deep peroneal nerve, muscle branches of the femoral nerve, and a nerve to the hamstring muscles--were studied with transganglionic transport of horseradish peroxidase (HRP) in the rat. The influence of varying the postoperative survival (3, 6, and 10 days) and of using wheat germ agglutinin-HRP conjugate (WGA-HRP), or HRP with dimethylsulfoxide (DMSO) instead of free HRP was studied for the gastrocnemius nerves. After 3 days' survival following application of HRP to the gastrocnemius nerves, fine granular labeling was found mainly in lamina V in L4-5, and coarse granular labeling was found in Clarke's column as far caudally as L2, and in laminae VI and VII predominantly in Th12-L2. After 6 or 10 days' survival, the fine labeling in lamina V was sparse or absent, whereas the coarse labeling appeared to remain or to be only slightly reduced in Clarke's column and in laminae VI and VII. No labeling suggestive of terminals was observed in laminae I-III from the gastrocnemius nerves. Except for sparse labeling in lamina I in some of the cases and some minor differences rostrocaudally, the spinal distribution of labeling was similar to that from the other nerves investigated. The distribution of labeling obtained after application of WGA-HRP or HRP with DMSO to the gastrocnemius nerves was very similar to that obtained with free HRP after 3 days' survival. The results indicate that the spinal cord projections of hindlimb muscle nerves in the rat distribute mainly in the deep part of the dorsal horn and in the intermediate zone. Furthermore, the lack of labeling suggestive of terminals in laminae I-III from the gastrocnemius nerves suggests, in conflict with earlier findings in the cat, that primary afferent fibers from muscles do not necessarily terminate in these laminae in the rat. The results suggest, furthermore, that fine granular labeling found in lamina V represents fine-calibered afferent fibers. Finally, the similar spinal projection patterns of the different muscle nerves investigated suggest either a less developed or an essentially different somatotopic organization for muscle afferents compared to cutaneous afferents, as revealed in earlier studies.     

Chronic peripheral nerve section results in a rearrangement of the central axonal arborizations of axotomized A beta primary afferent neurons in the rat spinal cord

In order to investigate the reorganization of the neuropil of the dorsal horn following peripheral nerve injury, the central terminal arborizations of 35 A beta primary afferent neurons, chronically injured by a cut and ligation of the sural nerve 6–12 weeks previously, were studied by the intra-axonal injection of horseradish peroxidase. Their morphology was compared to 13 intact sural nerve hair follicle afferents. Following axotomy, three kinds of morphological abnormalities were observed in the collateral arbors of the 26 afferents that were hair follicle-like. Atrophy with thin stem axons and reduced terminal branch patterns with few boutons was seen in 5 afferents. Sprouting of bouton-containing terminals into lamina I and IIo was found in 8 afferents. Finally, abnormal arborization patterns in the deeper laminae were observed in 29% of the collateral arbors. Changes included the loss in some arbors of a flame-shaped appearance, which is characteristic of hair follicle afferents, atypical branching patterns and ventrally directed axons producing wider and deeper arbors, compared to normal. Axotomy also caused a disruption of the normal somatotopic organizaiton of sural nerve A beta afferents. This disruption manifested as a variability in the normally mediolaterally restricted terminal sheet, with a consequent loss of the strict somatotopic register in the rostrocaudal direction. Damage to the peripheral axon of A beta primary afferents induces a structural reorganization of their central terminals in the dorsal horn of the spinal cord, which may modify sensory input to the central nervous system.     

Plasticity of acid phosphatase (FRAP) afferent terminal fields and of dorsal horn cell growth in the neonatal rat

Peripheral nerve section results in depletion of fluoride-resistant acid phosphatase (FRAP) from the nerve terminals in the dorsal horn of the spinal cord (Schoenen et al., '68) and this has been used in the past to map the termination field of individual nerves (Rustioni et al., '71; Devor and Claman, '80). In the present study we show that a similar central depletion occurs following sciatic nerve section or crush in neonatal rats. Unlike adults, however, the area of depletion is rapidly filled by sprouting of FRAP-containing afferent terminals from nearby intact peripheral nerves. The sprouting is extensive but never completely fills the depleted area. After nerve crush there is some recovery of FRAP from the sciatic nerve terminals themselves as well as from nearby nerve terminals. The source of recovered FRAP is demonstrated by resectioning or recrushing the nerves. The sprouting occurred when the sciatic was injured on day 1 but failed to take place when the injury was applied on or after day 10. Sciatic nerve section on day 1 also produces marked growth retardation of the ipsilateral dorsal horn gray matter that becomes more apparent as the rat matures. Nerve crush produces a less marked shrinkage that is slower in onset. If the nerve is crushed repeatedly, however, so that regeneration is prevented, the shrinkage is analogous to that following nerve section. No shrinkage occurs if the nerve is cut or crushed on day 10. The results show that separation of the spinal cord from its peripheral input at a critical stage in development results in disruption of the somatotopic organization of the C fibre afferent input to the dorsal horn and in slowing of growth of the dorsal horn gray matter.     

Difference in central projection of primary afferents innervating facial and intraoral structures in the rat

Transganglionic transport of horseradish peroxidase-wheat germ agglutinin conjugate was used to study the central projection of primary afferent neurons innervating facial and intraoral structures. The examined primary neurons innervating the facial structures were those comprising the frontal and zygomaticofacial nerves and those innervating the cornea, while the primary neurons innervating the intraoral structures included those innervating the mandibular incisor and molar tooth pulps and those comprising the palatine nerve. The primary afferents innervating the facial structures project to the lateral or ventral parts of the trigeminal principal, oral and interpolar subnuclei, and to the rostral cervical spinal dorsal horn across laminae I through V, with a greater proportion being directed to the spinal dorsal horn. The primary afferents innervating the intraoral structures terminate in the dorsomedial subdivisions of the trigeminal principal, oral and interpolar subnuclei, and in laminae I, II, and V of the medial medullary dorsal horn, with a much denser projection being distributed to the rostral subnuclei. In addition to the above brain stem trigeminal sensory nuclear complex, they project to the supratrigeminal nucleus, caudal solitary tract nucleus, and paratrigeminal nucleus. These observations agree with previously reported data that the central projection of trigeminal nerve is organized in different manners for the facial and intraoral structures. Furthermore, the present findings in conjunction with our previous studies clarify that the central projection of primary afferents from the facial skin is organized in a clear somatotopic fashion and that the terminal fields of primary afferents from the intraoral structures extensively overlap in the brain stem trigeminal nuclear complex particularly in its rostral subdivisions. The central mechanism of trigeminal nociception is discussed with particular respect to its difference between the facial and intraoral structures.     

Anterograde transport of horseradish-peroxidase conjugated isolectin B4 from Griffonia simplicifolia I in spinal primary sensory neurons of the rat

Anterograde transport of the isolectin B4 from Griffonia simplicifolia I (B4) conjugated to horseradish peroxidase (HRP) was investigated in rat somatic and visceral primary sensory neurons at different spinal levels. Injection of B4-HRP into the L5 dorsal root ganglion (DRG) resulted in labelling in the sural nerve, but not in the gastrocnemius nerves. Free nerve endings and lanceolate-like nerve endings were labelled in the lateral hindpaw skin. Labelled fibres were also observed in the greater splanchnic nerve following B4-HRP injection into the T10–11 DRGs. Electron microscopic examination of the labelled nerves showed that B4-HRP labelled exclusively unmyelinated axons. In the spinal cord, labelling was observed in the superficial dorsal horn, and additionally, although much more sparse, in the medial and lateral collateral projections following injections into the T10–11 DRGs. The results suggest that B4-HRP should be a suitable anterograde tracer of unmyelinated cutaneous and splanchnic but not muscle primary afferent fibres.     

Effect of nerve section on the spinal distribution of neighboring nerves

The spinal cord distribution of axonal terminals of peripheral nerves that innervate the skin of the upper medial thigh was examined in rats using transganglionic transport of horseradish peroxidase (HRP) and wheat-germ agglutinin-conjugated HRP (WGA-HRP). Chronic transection of the sciatic nerve or both the sciatic and saphenous nerves did not alter this distribution. Therefore, long-distance sprouting of intact 'thigh nerve' afferents in the dorsal horn is apparently not the mechanism whereby spinal dorsal horn neurons deafferented by sciatic and saphenous neurectomy, gain novel receptive fields in the cutaneous distribution of neighbouring intact nerves of the thigh.     

Topographic organization of central terminal region of different sensory branches of the rat mandibular nerve

The central projection of primary neurons comprising the auriculotemporal nerve, cutaneous branch of the mylohyoid nerve, inferior alveolar nerve, mental nerve, lingual nerve, and buccal nerve was investigated using transganglionic transport of HRP in young rats. In view of the topographic organization of central projection fields, the nerves were divided into two groups; i.e., those projecting to the dorsolateral margin of the trigeminal nucleus principalis, subnucleus oralis, and interpolaris (the auriculotemporal, mylohyoid, and mental nerves) and those projecting more medially (the inferior alveolar, lingual, and buccal nerves). The former group of nerves projected more caudally than the latter in the medullary and spinal dorsal horn complex rostral to the 3rd cervical segment, in general. Furthermore, the latter group projected to the nucleus of the solitary tract and the supratrigeminal and paratrigeminal nuclei, whereas the other nerves did not. The data indicate the following points: Primary neurons innervating the intraoral structures terminate medial (in trigeminal nucleus principalis and subnucleus oralis) and ventral (in subnucleus interpolaris) to the terminal fields of those innervating the facial skin. Primary neurons innervating the intraoral structures project to the nucleus of the solitary tract and the supra- and paratrigeminal nuclei, whereas those innervating the facial skin do not. Primary neurons innervating the periphery of the face project to the spinal dorsal horn and those innervating the intra/perioral region project to medullary dorsal horn, though this segregation from the medulla to the 3rd cervical segment is relatively loose. Only those trigeminal primary neurons, whose receptive fields extend to or beyond the midline, project to the contralateral dorsal horn from the medulla to the 3rd cervical segment.     

Spinal cord projections of the rat main forelimb nerves, studied by transganglionic transport of WGA-HRP and by the disappearance of acid phosphatase.

The spinal cord projections of the 3 main forelimb nerves-median, radial and ulnar, were studied in the rat dorsal horn with transganglionic transport of wheat germ agglutinin-horseradish peroxidase (WGA-HRP), or using the disappearance of fluoride resistant acid phosphatase (FRAP) after nerve section. The projection patterns in lamina II were similar following the two procedures. The median and the radial nerve fibers projected to the medial and the intermediate thirds, respectively, of the dorsal horn lamina II in spinal cord segments C4-C8. The ulnar nerve projected to segments C6-C8 between the areas occupied by the other two nerves. The FRAP method also showed that the lateral part of lamina II, which was not filled by radial nerve fibers, received the projections from the dorsal cutaneous branches of cervical spinal nerves. In addition, FRAP disappeared from the medial end of segment T1 after skin incisions extending from the medial brachium to the axilla, which seemed due to severance of the cutaneous branchlets of the lateral anterior thoracic nerve. The FRAP procedure is thus sensitive enough to detect fibers in lamina II arising from small peripheral nerves, and may be used as an alternative to the anterograde tracing methods whenever there are no overlapping projections.     

Somatotopic organization of cutaneous afferent terminals and dorsal horn neuronal receptive fields in the superficial and deep laminae of the rat lumbar spinal cord

The somatotopic organization of A- and C-afferent fibre terminals in the dorsal horn of the rat lumbar spinal cord was compared with the spatial location of second-order dorsal horn neuronal mechanoreceptive fields. The central terminal fields of the sural, saphenous, and tibial nerve were mapped by labelling the nerves with horseradish peroxidase (HRP). A previous study used the transganglionic transport of wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP) to produce a somatotopic map of high-threshold C-fibre terminal fields in lamina II (Swett and Woolf: J. Comp. Neurol. 231:66-77, '85). In the present study the terminal fields of low-threshold A beta afferents that terminate in laminae III and IV were mapped by using unconjugated HRP at prolonged survival times (72 hours). Unfixed tissue was used to increase the sensitivity of the tetramethylbenzidine reaction, thus allowing these afferent terminals to be clearly seen. The general spatial arrangement of the terminal fields in laminae III/IV closely resembled that found in lamina II in the mediolateral and rostrocaudal planes but because of a dorsoventral obliquity of the afferent terminals, the superficial and deeper fields are not in strict vertical register. The input to laminae II-IV of the dorsal horn may therefore be viewed as two horizontally arranged sheets of afferent terminals both accurately representing the skin surface, the more superficial sheet representing the high-threshold C-afferents and the deeper sheet, low-threshold A-beta afferents. The spatial organization of high-threshold A-delta afferents in laminae I and V appears to be quite different, with a transverse rather than a longitudinal orientation. To study dorsal horn cell receptive field organization two single units with mechanoreceptive fields were recorded extracellularly in each of 87 vertical tracks in the lumbar spinal cord, one unit in the superficial dorsal horn and the second in the deep dorsal horn. In general the somatotopic organization of the receptive fields of both sets of units followed that of the afferent terminal fields but there were cells with receptive fields that were anomalous relative to the recording site. No evidence of any vertical relation or columnar arrangement in receptive field size, threshold, or location on the body surface was found when comparing the two units in a pair. Furthermore, no laminar functional specialization was found, the majority of neurones having both low- and high-threshold inputs.(ABSTRACT TRUNCATED AT 400 WORDS)     

Spinal cord afferent systems containing the nerve terminal protein NT75

In the adult spinal cord, immunocytochemical staining for NT75 is concentrated in nerve terminals in the superficial laminae of the dorsal horn. Deeper laminae of the dorsal horn contain moderate immunocytochemical labeling, but the ventral horn is only sparsely stained. The origin of spinal nerve terminals containing NT75 was investigated with lesion techniques, colchicine treatment, and retrograde tracing in combination with immunocytochemical staining. Primary afferent neurons express NT75 immunoreactivity and account for most of the dense staining in the superficial dorsal horn and part of the labeling in the deeper laminae. It was found that corticospinal and virtually all brainstem neurons with descending projections to the spinal cord also express NT75 immunoreactivity, including those terminating in the ventral horn. Colchicine treatment of the spinal cord also resulted in NT75 staining in most, if not all, spinal neurons. It appears that neurons in all three major sources of spinal afferents (primary sensory, descending, and intrinsic systems) can express NT75 immunoreactivity, but that some neurons normally contain higher levels of the protein in their nerve terminals. Previous analysis of developing spinal cord has shown widespread, dense NT75 labeling throughout the spinal gray in the early postnatal period, which later becomes restricted to the adult pattern. These studies support the hypothesis that many spinal pathways express high levels of NT75 immunoreactivity during development, but that only certain pathways maintain high levels in the adult. © 1993 Wiley-Liss, Inc.     

Morphology and somatotopy of the central arborizations of rapidly adapting glabrous skin afferents in the rat lumbar spinal cord

The central arborizations in the dorsal horn of the spinal cord of 23 rapidly adapting (RA) A-beta primary afferent neurons innervating different regions of the glabrous skin of the hindpaw were studied by the intra-axonal injection of horseradish peroxidase in adult rats. A total of 284 arbors of the complex, simple, and blind-ending variety were recovered. The arbors of RA afferents innervating the toes, paw pads, and non-pad hindpaw differed from each other in branch pattern and dimensions. The simple and complex arbors, which are both bouton-containing, were distributed mainly in laminae III–V, although some complex arbors projected dorsally into lamina IIi. The hindpaw glabrous skin afferent terminals were located in the lumbar enlargement from caudal L3 to rostral L6. A crude somatotopic organization was observed such that toes 1–5 were represented successively in more caudal positions from mid-L4 to caudal L5. The paw pads were organized in a rostrocaudal sequence moving from the paw pads proximal to toe 1 across the foot to the paw pads proximal to toe 5, from caudal L3 to mid-L5. Non-pad hindpaw afferents were located in caudal L5. Overlap between toe, paw pad and non-pad afferent central fields was present, however, and the central terminals of afferents with non-adjacent peripheral receptive fields were shown to occupy the same region of the dorsal horn. © 1993 Wiley-Liss, Inc.