Convergence of forelimb afferent actions on C7-Th1 propriospinal neurones bilaterally projecting to sacral segments of the cat spinal cord

Propriospinal neurones located in the cervical enlargement and projecting bilaterally to sacral segments of the spinal cord were investigated electrophysiologically in eleven deeply anaesthetized cats. Excitatory or inhibitory postsynaptic potentials from forelimb afferents were recorded following stimulation of deep radial (DR), superficial radial (SR), median (Med) and ulnar (Uln) nerves. 26 cells were recorded from C7, 22 from C8 and 3 from Th1 segments. The majority of the cells were located in the Rexed's laminae VIII and the medial part of the lamina VII. In 10 cases no afferent input from the forelimb afferents was found. In the remaining neurones effects were evoked mostly from DR (88%) and Med (63%), less often from SR (46%) and Uln (46%). Inhibitory actions were more frequent than excitatory. The highest number of IPSPs was evoked from high threshold flexor reflex afferents (FRA)--all connections were polysynaptic. However, inhibitory actions were often evoked from group I or II muscle afferents (polysynaptic or disynaptic) and, less frequently, from cutaneous afferents (mostly polysynaptic). Di- or polysynaptic IPSPs often accompanied monosynaptic EPSPs from group I or II muscle afferents. Disynaptic or polysynaptic EPSPs from muscle and cutaneous afferents were also recorded in many neurones, while polysynaptic EPSPs from FRA were observed only exceptionally. Various patterns of convergence in individual neuronal subpopulations indicate that they integrate different types of the afferent input from various muscle and cutaneous receptors of the distal forelimb. They transmit this information to motor centers controlling hind limb muscles, forming a part of the system contributing to the process of coordination of movements of fore--and hind--limbs.     

Direct and relayed projection of periodontal receptor afferents to the cerebellum in the ferret

Field potentials in the cerebellar cortex of the ferret have been studied in response to stimulation of alveolar, muscular and cutaneous branches of the trigeminal nerve. Responses from the alveolar nerves are unusual in their very short latency. Evidence based on latency analysis, frequency following and comparison with other well-known inputs supports the view that the earliest field potentials are due to direct, unrelayed afferents, which terminate as mossy fibres. There is, in addition, a monosynaptically relayed afferent path via mossy fibres. The alveolar nerve afferents concerned with the direct projection are shown to come from periodontal mechanoreceptors and not from cutaneous receptors. No such connections are found from jaw-muscle spindle afferents. The direct and relayed periodontal pathways are both ipsilateral and crossed. They terminate in the cerebellar cortex in the parvermal region of lobules IV, V and VI. The functional significance of the direct periodontal afferent projection is considered particularly in the light of parallels with the vestibular system, which also has direct and relayed cerebellar projections.     

Proprioceptive afferent fibers in the cranial nerves III, IV and VI.

The purpose of the present works was to clarify whether the cranial nerves III, IV and VI carry proprioceptive afferent fibres from the extrinsic ocular muscles. In sheep the picture is now clear. The cranial nerves III, IV and VI carry many large proprioceptive fibres (12-16 micrometer) to the central nervous system. These nerves also contain many small fibres of the y-range (2-6 micrometer) which innervate the intrafusal muscle fibres in the spindles. In man the picture is still vague: most of the spindles are not typical, the large proprioceptive fibres (12-16 micrometer) and the small y-fibres (2-6 micrometer) are very few in the cranial nerves III, IV and VI. It is to be concluded that in sheep the cranial nerves III, IV and VI are not purely motor nerves to the extrinsic ocular muscles, but they also carry many of the large fibres of the proprioceptive function. In man, such large fibres are not found and the pathway of proprioceptive afferents from the orbital muscles is still not certain.     

Primary afferent depolarization of C fibres in the spinal cord of the cat

The excitability of primary afferent terminals of cutaneous C fibres was tested in the spinal cord of decerebrated cats. C fibre terminal excitability was decreased in the spinal state, and increased by conditioning volleys that activated only A fibres of another cutaneous nerve and by stimulating hair mechanically. It is suggested that C fibre input and therefore nociceptive information to the central nervous system is susceptible to presynaptic control by segmental and suprasegmental mechanisms.     

Study of tonic and evoked electrical activity in efferent fibers of the sympathetic nerve in white rats

In anaesthetised Wistar rats, electrical sympathetic activity and a somatosympathetic reflex in the cervical sympathetic trunk elicited by a single electrical shock to forelimb or hindlimb afferent nerves, were recorded. The spontaneous activity was shown to conform with the pulse and respiratory waves of arterial pressure. Somatosympathetic reflex consists of early and late discharges evoked by somatic myelinated afferent fibres stimulation, and C-response elicited by stimulation of unmyelinated afferent fibres in spinal nerves.     

The effect of stimulation of Golgi tendon organs and spindle receptors from hindlimb extensor muscles on supraspinal descending inhibitory mechanisms

Experiments were performed in precollicular decerebrate cats to investigate whether proprioceptive volleys originating from Golgi tendon organs and muscle spindles may activate supraspinal descending inhibitory mechanisms. Conditioning stimulation of the distal stump of ventral root filaments of L7 or S1 leading to isometric contraction of the gastrocnemius-soleus (GS) muscle inhibited the monosynaptic reflex elicited by stimulation of the ipsilateral plantaris-flexor digitorum and hallucis longus (Pl-FDHL) nerve. The amount and the time course of this Golgi inhibition were greatly increased by direct cross-excitation of the intramuscular branches of the group Ia afferents due to ephaptic stimulation of the sensory fibers, which occurred when a large number of a fibers had been synchronously activated. The postsynaptic and the presynaptic nature of these inhibitory effects, as well as their segmental origin, have been discussed. In no instance, however, did the stimulation of Golgi tendon organs elicit any late inhibition of the test monosynaptic reflex, which could be attributed to a spino-bulbo-spinal (SBS) reflex. Conditioning stimulation of both primary and secondary endings of muscle spindles, induced by dynamic stretch of the lateral gastrocnemius-soleus (LGS) muscle, was unable to elicit any late inhibition of the medial gastrocnemius (MG) monosynaptic reflex. The only changes observed in this experimental condition were a facilitation of the test reflex during the dynamic stretch of the LGS, followed at the end of the stimulus by a prolonged depression. These effects however were due to segmental interactions, since they persisted after postbrachial section of the spinal cord. Intravenous injection of an anticholinesterase, at a dose which greatly potentiated the SBS reflex inhibition produced by conditioning stimulation of the dorsal root L6, did not alter the changes in time course of the test reflex induced either by muscle contraction or by dynamic muscle stretch. Conditioning stimulation of a muscle nerve activated the supraspinal descending mechanism responsible for the inhibitory phase of the SBS reflex only when the high threshold group III muscle afferents (innervating pressure-pain receptors) had been recruited by the electric stimulus. This finding contrasts with the great availability of the system to the low threshold cutaneous afferents. The proprioceptive afferent volleys originating from Golgi tendon organs as well as from both primary and secondary endings of muscle spindles, contrary to the cutaneous and the high threshold muscle afferent volleys, were apparently unable to elicit not only a SBS reflex inhibition, but also any delayed facilitation of monosynaptic extensor reflexes attributable to inhibition of the cerebellar Purkinje cells.     

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.     

刺猬胸前神经传入活动的来源及皮层代表区分布

为探讨冬眠刺猬皮肌的紧张性控制,本工作研究了支配皮肌的胸前神经(VTN)的传入活动的来源和皮层代表区的分布。VTN的传入冲动来自皮肌本体感受器,传入纤维径C_6—T_2背根入脊髓,与同部位的皮肤感觉相分离,后者经相应节段的皮神经传入。电刺激VTN引起的皮层诱发电位反应位于新皮层外侧面的中间部,相当于Woolsey的S-Ⅱ区内,与桡神经和坐骨神经的代表区有重叠,而在S-Ⅰ区没有记录到反应。     

Interaction between segmental and descending intersegmental reflexes in lumbar motoneurons

The effects of segmental reflexes on descending intersegmental reflexes to stimulation of forelimb afferents were studied in anesthetized cats by recording postsynaptic responses from single motoneurons. Interaction between these influences was found to be reciprocal in character for groups of neurons with primary connections with afferents of the superficial and deep branches of the peroneal nerve and afferents of the nerve to the gastrocnemius muscle. Excitatory postsynaptic responses arising in groups of motoneurons of the peroneal nerve to stimulation of forelimb afferents underwent profound and prolonged inhibition during conditioning stimulation of afferents in the deep and superficial peroneal nerves. Activation of segmental afferents during conditioning stimulation of the gastrocnemius nerve was accompanied by inhibition of excitatory intersegmental responses and deinhibition of inhibitory responses in motoneurons of the gastrocnemius muscle. Segmental inhibition of intersegmental descending impulse activity appeared in the interneuron system of the segmental reflex centers connecting the descending propriospinal tracts with the motoneurons of these centers.I. P. Pavlov Institute of Physiology, Academy of Sciences of the USSR, Leningrad. Translated from Neirofiziologiya, Vol. 4, No. 2, pp. 16872-175, March–April, 1972.     

Effectiveness of ascending synaptic influences from various spinal funiculi on reticulospinal neurons in cats

Activity of reticulospinal neurons evoked by stimulation of the ventral, ventrolateral, dorsolateral, and dorsal funiculi of the spinal cord was recorded extracellularly in cats anesthetized with chloralose. Responses of 57 reticulospinal neurons, of which 22 projected into the ventral funiculus, 20 into the ventrolateral, and 15 into the dorsolateral, were studied. The functional properties (conduction velocity and refractory period) and the location of the neurons of the above-mentioned groups in the medulla did not differ appreciably. The most effective synaptic activation of all neurons was observed during stimulation of the dorsal and dorsolateral funiculi. Responses to stimulation of the dorsal funiculus had the lowest threshold. These responses arose in reticulospinal neurons of the ventral and ventrolateral funiculi after the shortest latent period. The effectiveness of synaptic influences from the dorsal and dorsolateral funiculi was identical in the group of neurons of the dorsolateral funiculus. Correlation between activity evoked by stimulation of the dorsal funiculus in reticulospinal neurons and peripheral nerves indicated that the responses appeared in these cells to stimulation of muscular (groups I and II) and cutaneous (group II) afferent fibers. The results indicate that impulses from low-threshold muscular and cutaneous afferents, which effectively activate reticulospinal neurons, are transmitted along ascending pathways of the dorsal funiculi.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 11, No. 3, pp. 254–263, May–June, 1979.     

Primary afferent depolarization in the cervical enlargement of cats evoked by activation of cervical sensory nerves

Depolarization of primary afferent terminals in the cervical enlargement of the spinal cord evoked by activation of sensory nerves of the upper cervical segments (C₂) was studied in cats anesthetized with pentobarbital. It was shown that low-threshold muscular and high-threshold cutaneous afferents of nerves of the forelimb were depolarized most strongly. Parallel with this depolarization, prolonged (over 0.5 sec) inhibition of the monosynaptic and polysynaptic flexor reflex developed. It is suggested that these influences are transmitted via pathways running in the posterior and lateral white columns. The results are discussed in connection with regulation of postural motor activity in vertebrates.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 14, No. 2, pp. 190–197, March–April, 1982.     

Relevance of the callosal transfer in defining the peripheral reactivity of somesthetic cortical neurones

Experiments were performed in 13 chloralose-anaesthetized, curarized cat preparations (monitoring of rectal temperature, heart rate, expired pCO2 and EEG), in order to ascertain whether, and to what extent, the reactivity to ipsilateral skin shocks of the neurones of the anterior ectosylvian and anterior suprasylvian gyri (AEG and ASG, respectively) is dependent on the callosal output of the somatosensory areas of the contralateral hemisphere. Indeed, we knew from previous experiments that a high proportion of AEG and ASG neurones having bilateral peripheral receptive fields (PRFs) can be excited by direct stimulation of the contralateral homonymous areas, and that the callosal fibres originating in the latter carry somesthetic impulses related to contralateral PRFs. A preliminary analysis was carried out on the amplitude and latency relationships between the evoked potentials (EP) recorded simultaneously from the two hemispheres and from the corpus callosum (CC) following stimulation of the forepaw of one side. The results obtained showed good correlations between the onset and development of the EPs picked up from the hemisphere ipsilateral to the stimulated skin, on the one hand, and onset and development of the EPs recorded from the contralateral hemisphere and the corpus callosum, on the other. At a further stage of the experiments, the EPs elicited upon ipsilateral and contralateral skin shocks in the AEG-ASG area have been recorded and averaged before, during and after the reversible inactivation of callosal somesthetic transmission. This was achieved by applying polarizing currents (0.2-1 mA) to the rostral portion of the CC, adequacy and reversibility of this method having been tested by observing, respectively, suppression and prompt restoration of transcallosal EPs and of the asynaptic spiking produced by cortical cells when antidromically invaded from contralateral homotopic cortex. It was seen that during CC blockade the EPs elicited in the AEG-ASG areas did not show any change either in amplitude or time-course if brought about by contralateral peripheral stimulation, whereas those evoked by ipsilateral skin shocks exhibited significant reduction, which was related to the strength of CC polarization and to the reduction of transcallosal EPs. In control experiments similar effects were observed after ablation of somatosensory areas of the hemisphere which send off somesthetic callosal impulses, whereas strychninization of these areas caused effects opposite in sign, i.e., enhancement of the ipsilateral but not of the contralateral EPs in the areas of the untreated hemisphere. By testing the effects of CC polarization on single AEG-ASG neurones, it was observed that the responses of units linked only with contralateral PRFs (Group I; 7 units tested) were unaffected by callosal polarization. The discharges of neurones provided with wide and bilateral PRFs (Group II; 27 units tested) were not affected if elicited by contralateral PRF shocks but were deeply impaired (in 11 neurones out the 27) when provoked by ipsilateral PRF stimulation. The effect consisted chiefly of the disappearance of the first high peak of the PSTHs, and when recording intracellularly graded events, it was mirrored by a large decrease of the postsynaptic excitatory potentials elicited in Group II neurones by ipsilateral PRF shocks. A late scattered histographic component was identified in the PSTHs of such cells, which did not appear to be significantly altered during CC blockade. These effects were observed on the ipsilateral responses of 11 out of the 27 Group II neurones so tested whereas the ipsilateral PSTHs of the remaining 16 Group II neurones either did not undergo significant changes during the callosal blockade or escaped evaluation because of high spontaneous shifts of neural responsiveness. The results are discussed mainly with a view to the possible functional role of the specific somesthetic callosal fibres in defining ipsilateral reactivity for the wide-field cells of the AEG-ASG area.     

On the nature of the afferent fibers of oculomotor nerve

The oculogyric nerves contain afferent fibers originating from the ophthalmic territory, the somata of which are located in the ipsilateral semilunar ganglion. These primary sensory neurons project to the Subnucleus Gelatinosus of the Nucleus Caudalis Trigemini, where they make presynaptic contact with the central endings of the primary trigeminal afferents running in the fifth cranial nerve. After complete section of the trigeminal root, the antidromic volleys elicited in the trunk of the third cranial nerve by stimulating SG of NCT consisted of two waves belonging to the A delta and C groups. The area of both components of the antidromic volleys decreased both after bradykinin and hystamine injection into the corresponding cutaneous region and after thermic stimulation of the ipsilateral trigeminal ophthalmic territory. The reduction of such potentials can be explained in terms of collision between the antidromic volleys and those elicited orthodromically by chemical and thermic stimulation. Also, capsaicin applied on the nerve induced an immediate increase, followed by a long lasting decrease, of orthodromic evoked response area. These findings bring further support to the nociceptive nature of the afferent fibers running into the oculomotor nerve.     

Mechanisms of pain arising from articular tissues

This paper reviews the peripheral and central neural mechanisms underlying pain from articular tissues innervated by spinal and trigeminal afferents. The paper especially addresses trigeminal mechanisms related to pain from the temporomandibular joint and its associated craniofacial musculature. Recent studies have shown the existence of articular nociceptive primary afferents that project to the spinal cord dorsal horn and trigeminal brainstem complex. A particular feature of most neurones receiving these deep nociceptive afferent inputs is their responsivity also to cutaneous nociceptive afferent inputs. This suggests the involvement of these neurones not only in the detection of acute articular pain, but also in the hyperalgesia and poor localization, spread, and referral of pain that characterize many painful conditions of joints and other deep structures. While only limited information is available on related higher brain centre mechanisms, convergence and interaction between cutaneous and deep afferent inputs also seem to be a characteristic of somatosensory neurones in the thalamus and somatosensory cerebral cortex. Muscle and autonomic reflexes may be induced by such deep noxious stimuli, but the functional significance of some of these effects (e.g., in relation to clinical concepts of myofascial dysfunction) requires further study in more appropriate functional settings.     

Synaptic potentials of motoneurons of the masseter muscle

Postsynaptic potentials of 93 motoneurons of the masseter muscle evoked by stimulation of different branches of the trigeminal nerve were studied. Stimulation of the most excitable afferent fibers of the motor nerve of the masseter muscle evoked monosynaptic EPSPs with a latent period of 1.2–2.0 msec, changing into action potentials when the strength of stimulation was increased. A further increase in the strength of stimulation produced an antidromic action potential in the motoneurons with a latent period of 0.9 msec. In some motoneurons polysynaptic EPSPs and action potentials developed following stimulation of the motor nerve to the masseter muscle. The ascending phase of synaptic and antidromic action potentials was subdivided into IS and SD components, while the descending phase ended with definite depolarization and hyperpolarization after-potentials. Stimulation of cutaneous branches of the trigeminal nerve, and also of the motor nerve of the antagonist muscle (digastric) evoked IPSPs with a latent period of 2.7–3.5 msec in motoneurons of the masseter muscle. These results indicate the existence of functional connections between motoneurons of the masseter muscle and its proprioceptive afferent fibers, and also with proprioceptive afferent fibers of the antagonist muscle and cutaneous afferent fibers.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 1, No. 3, pp. 262–268, November–December, 1969.     

Primary afferent depolarization induced by activation of intersegmental connections of the substantia gelatinosa in the cat spinal cord

Depolarization of primary afferent terminals induced by selective activation of intersegmental connections of the substantia gelatinosa was investigated in cats anesthetized with pentobarbital. Depolarization was found to develop most rapidly in fibers of high-threshold muscular and cutaneous sensory nerves, but it was present only to a very slight degree in fibers of group Ia muscular afferents. It is suggested that the spread of activity inducing depolarization takes place in the substantia gelatinosa along a chain of excessively excited neurons. The possible role of primary afferent depolarization as a factor stabilizing coordinated activity of spinal neurons is discussed.     

Cortical Presynaptic Control of Dorsal Horn C–Afferents in the Rat

Lamina 5 sensorimotor cortex pyramidal neurons project to the spinal cord, participating in the modulation of several modalities of information transmission. A well-studied mechanism by which the corticospinal projection modulates sensory information is primary afferent depolarization, which has been characterized in fast muscular and cutaneous, but not in slow-conducting nociceptive skin afferents. Here we investigated whether the inhibition of nociceptive sensory information, produced by activation of the sensorimotor cortex, involves a direct presynaptic modulation of C primary afferents. In anaesthetized male Wistar rats, we analyzed the effects of sensorimotor cortex activation on post tetanic potentiation (PTP) and the paired pulse ratio (PPR) of dorsal horn field potentials evoked by C–fiber stimulation in the sural (SU) and sciatic (SC) nerves. We also explored the time course of the excitability changes in nociceptive afferents produced by cortical stimulation. We observed that the development of PTP was completely blocked when C-fiber tetanic stimulation was paired with cortex stimulation. In addition, sensorimotor cortex activation by topical administration of bicuculline (BIC) produced a reduction in the amplitude of C–fiber responses, as well as an increase in the PPR. Furthermore, increases in the intraspinal excitability of slow-conducting fiber terminals, produced by sensorimotor cortex stimulation, were indicative of primary afferent depolarization. Topical administration of BIC in the spinal cord blocked the inhibition of C–fiber neuronal responses produced by cortical stimulation. Dorsal horn neurons responding to sensorimotor cortex stimulation also exhibited a peripheral receptive field and responded to stimulation of fast cutaneous myelinated fibers. Our results suggest that corticospinal inhibition of nociceptive responses is due in part to a modulation of the excitability of primary C–fibers by means of GABAergic inhibitory interneurons.     

A possible coordinating role for presynaptic inhibition

Conditioning stimuli were applied to the common peroneal or superficial peroneal nerve in acute experiments on anesthetized cats. Changes in the N₁-component of the dorsal cord potential evoked by stimulation of one of these nerves or of other nerves (tibial, deep peroneal) and changes in the amplitude of antidromic action potentials in the afferent fibers of these nerves were investigated. The degree of reinforcement of antidromic action potentials, reflecting the degree of depolarization of the afferent terminals, was found to be greater for the passive nerve than for the active to which the conditioning stimulus was applied. Inhibition of the N₁-component of the dorsal cord potential was deeper when a pair of stimuli was applied to two different nerves (under these conditions only the mechanism of presynaptic inhibition was activated) than when they were applied to the same nerve. It is concluded that presynaptic inhibition, by selectively controlling afferent volleys, can evidently play a coordinating role.     

电刺激大鼠脊神经皮支对远距离机械感受单位电活动的影响

实验采用分离神经细束的方法,观察逆行电刺激大鼠脊神经背侧皮支后,在相距较远的神经细束上记录到的Aδ和C类机械感受单位电活动的变化。刺激T9脊神经背侧皮支,在T12神经细束上记录到59.3%（16/27）的Aδ和71.2%(37/52）的C类单位在刺激后90～120s放电显著增加。刺激T8脊神经背侧皮支,在T12神经细束上记录到47.8%(11/23）的Aδ单位和36.6%(15/41）的C类单位在刺激后120～150s放电显著增加。大多数单位（18/23）的机械感受阈值在电刺激远距离脊神经背侧皮支后降低。结果表明,逆行电刺激外周感觉神经,可以使相距较远的Aδ和C类机械感受单位致敏,其传入放电增加。     

Cutaneous nerves of the embryonic chick wing do not develop in regions denuded of ectoderm

Peripheral nerves travel to their targets along precise routes, and it is likely that different cues provide guidance at different stages of the journey. In a developing chick limb, the cutaneous nerve fibres follow at first deep mixed nerve trunks, in company with motor axons; they branch from these trunks at predictable points and approach the skin; they then ramify profusely to form a plexus at a precisely defined depth beneath the ectoderm, at exactly the same level as the blood vascular plexus. To analyse the role of signals from the target patch of skin in regulating cutaneous nerve development, we have ablated patches of dorsal wing ectoderm using short-wave ultraviolet irradiation at E4 (embryonic day 4), approximately one day before nerves grow into the limb bud. The irradiated patches remain denuded of ectoderm for more than a week, by which time the cutaneous nerve plexus on the contralateral control side is well developed and can be revealed by whole-mount silver staining. Where the ectoderm has been ablated, no cutaneous nerve plexus forms, and the nerve branches that normally would have diverged from the neighbouring mixed nerve trunk to innervate the missing patch of skin are absent - ab initio, apparently. The routes of the mixed nerve trunks are not affected. Partial ablation of the territory of a cutaneous nerve branch often leads to loss of the whole nerve branch; the intact skin territory thus left vacant is invaded by ramifications from the remaining cutaneous branches, as expected if the normal extent of a cutaneous nerve's territory is regulated by competition. Where there is an ectodermal lesion, cutaneous innervation stops precisely at its boundary, even though the vascular plexus extends for some distance beyond this margin, beneath the denuded surface. The data suggest that the embryonic skin is required firstly to trigger divergence of cutaneous nerve branches from the mixed nerve trunks, and secondly, once the nerve fibres have reached the skin, to supply a trophic cue (probably NGF) encouraging growth of a plexus; at the same time, the embryonic skin generates a signal inhibiting nerves from approaching closer than about 70 microns to the surface.