Joint Afferents Research Articles

Age-related loss of knee joint afferents in mice.

Previous work in our laboratory revealed markedly different rates of age-related death of four monoaminergic neuronal populations in the C57BL/6 mouse. Although dorsal root ganglion neurons (DRGns) have been reported not to suffer similar age-related death in rodents, we determined if there is age-related death of the subpopulation of DRGns innervating the knee joints of C57BL/6 mice, which are known to develop degenerative arthritis with aging. The somata of dorsal root ganglion neurons innervating the mouse knee joint (KJ-DRGns) were identified by retrograde tracing with Fluoro-Gold (FG). Lumbar ganglia were serially sectioned and the numbers of FG-labelled KJ-DRGns counted at five ages encompassing the animal's life span. Changes in size of the total population of lumbar DRGns (L-DRGns) were estimated by counting nucleated somata from every fifth toluidine blue-stained serial section from the L3 and L4 lumbar ganglia at three different ages. Using a computer-assisted video morphometric technique somal areas were measured from random sections to determine the distribution of sizes of neurons in the KJ-DRGn and general lumbar DRGn populations at different ages. Counts of FG-labelled joint afferents were 238.5 +/- 80.3 (mean +/- SD) KJ-DRGns per knee at 2 months of age, declining to 103.2 +/- 20.1 by 24 months, representing a 57% loss over the average life span of the C57 mice. The loss occurred in two phases, with a rapid rate over the first 8 months of life and a more moderate rate of loss over the remaining months. L-DRGn numbers revealed a slower overall rate of loss in comparison to the KJ-DRGn population with an average 33.7% loss over the life span of this mouse. Somal size measurements revealed that the larger sizes of KJ-DRGns were lost over the first 8 months of life, with little change in the distribution of somal sizes thereafter. The distributions of sizes of the L-DRGn population did not change significantly over the life spans of the mice. The data provides evidence that the age-related loss of KJ-DRGns is significantly greater than DRGns in general, and may be particularly apparent in the population of larger sized presumed mechanoreceptor neurons. The loss of the KJ-DRGns is approximately reciprocal to the incidence rate of knee joint osteoarthritis reported for the C57BL/6 mice.

The effects of phorbol ester on slowly conducting afferents of the cat's knee joint

The effects of beta-phorbol 12,13-dibutyrate (PDBu) on the discharge properties of slowly conducting knee joint afferents (group III and group IV fibers) were studied to determine the role of protein kinase C in nociception. Extracellular single unit recordings were made from small filaments dissected from the medial articular nerve in cats anesthetized with alpha-chloralose. PDBu was applied intra-arterially close to the joint in concentrations of 10(-6) up to 10(-4) M. The afferents were classified as low-threshold and high-threshold units with regard to their sensitivity to passive noxious and innocuous movements of the knee joint. Following PDBu application, an excitation occurred in 28% of the group III and in 40% of the group IV fibers. An enhancement of responses to passive movements of the joint (sensitization) occurred in 37% of group III and 19% of group IV afferents. In summary, 37.5% of the low-threshold and 50% of the high-threshold fibers proved to be sensitive to PDBu. Most of the PDBu-positive units responded also to bradykinin, whereas only a few PDBu-positive units were sensitive to prostaglandin I2 and E2. We conclude from these results that, in a distinct population of slowly conducting joint afferents, protein kinase C is likely to be involved in the process of transduction. Thus, pain and hyperalgesia may be mediated at least partly by intracellular mechanisms that are linked to protein kinase C.

Spontaneous activity of cat spinal ganglion neurons in vivo

Recordings were made from L4-S1 spinal ganglion neurons of anesthetized cats while their associated dorsal root and the sciatic nerve were left intact, locally anesthetized, or locally anesthetized and sectioned. In all three experimental conditions spontaneous discharges were recorded. These discharges occurred in the absence of any electrical stimulation of the dorsal root or sciatic nerve, and were not due to peripheral exploration of receptive fields or sustained firing in joint or muscle afferents. The spontaneous discharges were relatively rhythmic, and their firing frequency ranged from 5 to 100 impulses per s. Interactions between spontaneous and electrically evoked discharges were observed that depended on the impulse's frequency of firing. High frequency discharges always abolished low frequency impulses regardless of whether these latter were spontaneous or evoked. Extra spikes and postspike events that followed impulses evoked by stimulation of the dorsal root or sciatic nerve were also recorded from some spinal ganglion neurons. These results suggest that spontaneous discharges may originate within the spinal ganglion itself, and that they can occur under normal circumstances.

Knee joint input into the peripheral region of the ventral posterior lateral nucleus of cat thalamus.

1. Experiments were carried out in chloralose-anesthetized cats to study the responses of neurons in the lateral thalamus to excitation of afferent fibres from the knee joint. 2. Single- and multi-unit recordings were made with tungsten electrodes in dorsoventral penetrations through the ventral posterior lateral nucleus (VPL) during electrical stimulation of the medial articular nerve (MAN) of the cat's knee joint at an intensity sufficient to excite slowly conducting unmyelinated fibers. The locations of the recording sites were verified by recovering electrolytic lesion sites in histological sections (Nissl and cytochrome oxidase staining). 3. The average earliest latency for excitation of thalamic responses was 19.1 +/- 8.5 (SD) ms (n = 50). The threshold for excitation of most thalamic units was found to correspond to peripheral joint afferent fibers of the A-delta group. 4. The majority of neurons responding to MAN stimulation were found to be dorsal or ventral to the low-threshold cutaneous hindlimb region of the lateral division of the VPL (stereotaxic coordinates: AP 9.0-11.5; ML 7.0-9.5). In the ventral periphery of the VPL, most neurons responding to MAN stimulation (11/14) were wide dynamic range (WDR) with a discrete cutaneous receptive field on the hindpaw digits. Six WDR neurons were found dorsal to the hindlimb VPL with a convergent receptive field on the hindlimb (but not hindpaw digits). No nociceptive-specific knee joint units were found. 5. Other neurons were found dorsal to the hindlimb VPL with large receptive fields often encompassing the whole contralateral leg, including skin and deep hindlimb structures, possibly in a region described as the dorsal portion of the posterior complex (POd). Some neurons were found with no receptive field. 6. This study provides the first observations on the responses of lateral thalamic neurons to stimulation of the MAN of the cat knee joint. These results demonstrate a central pathway conveying impulses from specific deep joint afferents of the MAN to the peripheral region of the VPL and overlying region known as the POd, regions implicated in the transmission of nociceptive information.

Contribution of joint and muscle afferents to position sense at the human proximal interphalangeal joint

Experiments were carried out to examine the perceived position of the human index finger about the proximal interphalangeal joint. In protocol I, the finger was moved from an intermediate position at velocities ranging from 200 deg/min to 2 deg/min, then held still at one of three positions. The subject's task was visually to align a finger silhouette that was coaxial with the joint to the kinaesthetically perceived position of the unseen finger. Judgements of position were found to be quite accurate, and unaffected by previous velocity. Protocol II showed that although the direction of joint displacements of 0.01 and 0.1 deg could not be detected at any velocity, 1 deg could be detected at 200 deg/min and 10 deg at 20 and 2 deg/min. In protocol III the finger was moved at 2 deg/min and maintained at either 105 or 175 deg. It was found that the position of the unanaesthetized finger was quite accurately known, but with digital nerve block, subjects clearly perceived the finger to be at the mid-position (approximately 130-150 deg). This suggests that the absence of joint and cutaneous afference is interpreted by the CNS as indicating mid-position. The slight bias of the sensed position towards the objective position shown by the results also indicates that muscle afferents can provide a crude signal related to joint position. This finding was further supported by the observation that splinting the distal interphalangeal joint into flexion resulted in flexion bias in the perceived angle of the proximal interphalangeal joint. Anaesthesia of the middle finger, thumb and distal portion of the index finger (leaving proximal joint unaffected), had little effect on position matching performance, suggesting that the large error in position sense during anaesthesia of the whole finger is due to loss of afference specifically related to the proximal interphalangeal joint, and not due to loss of non-specific facilitatory influences from cutaneous and joint afferents. The results argue for an important proprioceptive role for joint afferents at finger joints.

Kinaesthetic signals and muscle contraction.

Signals generated both peripherally and centrally contribute to the group of sensations termed kinaesthesia. Many experiments report sensations of position and movement under passive relaxed conditions without muscle contraction. However, kinaesthetic acuity is probably of greater functional value when subjects are active rather than passive and, accordingly, movement detection is markedly improved by muscular contraction. One mechanism contributing to this enhancement is likely to involve muscle spindle volleys. When identical microstimulation techniques are applied to skin, joint and muscle spindle endings innervating the hand, some cutaneous afferents and some joint afferents elicit a sensation, but activation of certain other cutaneous afferents and muscle spindle afferents rarely does. Activity in more than one muscle spindle afferent may be required for kinaesthetic sensations, whereas some single cutaneous and joint afferents may have a more 'secure' central projection.

Influence of peripheral afferents on cortical and spinal motoneuron excitability

The objective of this study was to establish to what extent muscle, cutaneous, and joint afferents alter the excitability of spinal and cortical motor neurons. This question was examined by studying the impact of electrical stimulation of the second and third digits, the median nerve at the wrist, and the recurrent thenar motor branch on the F/H and magneto-electrical cortical motor responses (MEPs) of the thenar muscles. The firing frequencies of single F/H motor unit action potentials were unaltered by the foregoing conditioning peripheral stimuli. MEPs conditioned by motor threshold stimulation of the median nerve at the wrist or the recurrent motor branch were significantly increased in size at conditioning to test intervals of 50 to 80 milliseconds. No significant change in MEP size resulted from conditioning stimulation of the digital nerves. We conclude that muscle afferents were primarily responsible for the increase in MEP size. Conditioning stimuli may allow examiners to assess central motor conduction where it would otherwise be impossible.

Tonic descending inhibition of spinal cord neurones driven by joint afferents in normal cats and in cats with an inflamed knee joint

In ten cats, single unit electrical activity was recorded in the lumbosacral spinal cord from neurones driven by stimulation of afferent fibres from the ipsilateral knee joint. Tonic descending inhibition (TDI) on the responses of these cells was measured as increases in resting and evoked activity of the neurones following reversible spinalization of the animals with a cold block at upper lumbar level. Acute inflammation of the knee joint was induced in five of the cats by the injection of kaolin and carrageenan into the joint. TDI was observed in 25 of 33 neurones recorded in normal animals (76%) and in 36 of 40 (90%) neurones recorded in animals with acute knee joint inflammation. In both kinds of preparation TDI was more pronounced in neurones recorded in the deep dorsal horn and in the ventral horn than in those recorded in the superficial dorsal horn. There was a tendency in the whole sample for TDI to be greater in neurones with input from inflamed knees. We conclude that the spinal processing of afferent information from joints is under tonic descending influences and that the amount of TDI can be altered during acute arthritis.

Substance P- and calcitonin gene-related peptide immunoreactivity in primary afferent neurons of the cat's knee joint

The distribution of substance P and calcitonin gene-related peptide was determined in primary afferent neurons of the medial and posterior articular nerve of the cat's knee joint. Perikarya of articular afferents were visualized by retrograde labelling with the fluorescent dye Fast Blue which was applied at the transected end of the peripheral nerves. Substance P was found in about 17% of labelled medial articular afferents and in about 16% of labelled posterior articular afferents, respectively, whereas calcitonin gene-related peptide was present in about 35 and 32% of the medial and posterior articular nerve cells, respectively. Taking into account that these neuropeptides are known to be co-localized, probably not more than one-third of the joint afferents contain substance P and/or calcitonin gene-related peptide. Quantification of cell diameters revealed that substance P was found only in small- or intermediate-sized perikarya (< 50 μm) indicating that this peptide is predominantly found in unmyelinated neurons. Calcitonin gene-related peptide was present mainly in small- and intermediate- but also in some large-sized neurons (> 50 μm) providing evidence that this peptide is found in unmyelinated and to a lesser extent in myelinated neurons. This is consistent with previous studies that show that substance P and calcitonin gene-related peptide are present primarily in unmyelinated and thinly myelinated primary afferents. When the portion of substance P-positive neurons of the medial articular nerve is compared to the number of articular afferents displaying a nociceptive function as determined in earlier electrophysiological studies, it can be calculated that at most 30% of the nociceptive-specific articular afferents contain this neuropeptide. Since this finding parallels observations in other combined electrophysiological and immunocytochemical investigations, these data support the idea that substance P may be a functionally important neuropeptide in a small particular subgroup of nociceptive articular afferents but is not an essential neuropeptide in all articular nociceptive nerve fibres.

Joint receptors modulate short and long latency muscle responses in the awake cat

Nerve cuff electrodes were chronically implanted around multiple peripheral nerves in adult cats, including the medial and posterior articular nerves (MAN and PAN) to the knee while EMG electrodes were implanted into seven hindlimb muscles. Randomized load perturbations producing mid-range knee flexions at varying angular velocities were subsequently applied to awake cats. Recordings were initially obtained with knee joint innervation intact and then after local anaesthetic or saline control solution was injected into the knee. Averaged neurogram and EMG responses to the imposed movements were utilized to assess the contribution of joint mechanoreceptor activity to the evoked muscle responses. Additionally, spike-triggered averaging techniques and peri-stimulus time histograms of single joint afferent units isolated from the articular nerve cuffs were utilized to characterize unitary joint receptor responses. The averaged whole nerve response to knee joint perturbations on each of the cuffed articular nerves revealed phasic increases in activity relative to constant background levels. The earliest phasic responses on the articular nerves were initiated at latencies that were too short to be voluntary, occurring in the short latency (reflex) period. Detectable joint receptors were not recruited until after the earliest excitatory responses of agonist/antagonist muscle pairs acting across the knee had occurred, presumably resulting in mechanical loading of the knee joint capsule and subsequent activation of articular mechanoreceptors. Introduction of local anaesthetic into the knee was accompanied by marked diminution in joint afferent activity. Perturbation-evoked muscle responses were characterized by increased activity above background levels in all seven muscles studied, including antagonist muscle pairs. Local anaesthetic-mediated loss of knee joint mechanoreceptor input altered the latency, amplitude and duration of EMG responses in each muscle. The effect of joint anaesthesia in the short latency period was a generalized decrease in all muscle responses relative to normal and saline controls. The loss of afferent input after joint anaesthesia was also associated with altered muscle responses during the long latency period, when both reflex and voluntary mechanisms could potentially contribute to the generation of EMG activity. Interestingly, long latency activity after joint anaesthesia was characterized by "unbalancing" in the EMG responses of some antagonist muscle pairs. This alteration of normal antagonist pair co-contraction patterns served to increase the magnitude of the imposed perturbations, rather than to bring the movements under control. Analysis of single joint afferents isolated from whole joint nerve recordings demonstrated that some joint afferent units were tonically active at quiescent, mid-range knee positions. Additionally, isolated afferents demonstrated different time courses of response to imposed perturbations.(ABSTRACT TRUNCATED AT 400 WORDS)

Contralaterally projecting lamina VIII interneurones in middle lumbar segments in the cat

Peripheral input to lamina VIII interneurones was investigated by using extracellular and/or intracellular records from them. The interneurones were located in the L4-L5 spinal segments and projected to contralateral motor nuclei in the L7 segment. They constituted a non-homogeneous population but their input from muscle afferents (mainly group II afferents of quadriceps, flexor digitorum longus and pretibial flexors and group I afferents of triceps surae and hamstring nerves) and from cutaneous and joint afferents resembled the input to ipsilaterally projecting laminae V–VII interneurones of the same segments rather than the input to more caudally located lamina VII interneurones. Since the ipsilaterally projecting laminae V–VII interneurones with such an input might be involved in locomotion, it is proposed that this is also the case for the contralaterally projecting lamina VIII midlumbar interneurones, especially those excited by stimuli applied in the cuneiform nucleus (mesencephalic locomotor region).

Fourier analysis of the relation between the discharge of quadriceps motor units and periodic mechanical stimulation of cat knee joint receptors

It has been widely assumed that joint receptors contribute to the reflex regulation of movement and posture, although there have been few clear demonstrations of joint-mediated reflex actions on alpha-motoneurones other than those attributable to flexor reflex afferents. The present study extends our recent work on joint-mediated reflexes by using Fourier analysis of spike train interactions to demonstrate that restricted mechanical activation of a small number (one to five) of knee joint receptors by localized indentation of the joint capsule can modify the firing pattern of quadriceps motoneurones in decerebrated cats. The modulation of this discharge pattern can be reversibly abolished by application of droplets of lignocaine directly to the joint nerve and consequently can be attributed unambiguously to articular afferents. Activation of single joint afferents could on occasion produce changes in motor unit firing patterns, but usually activation of two or more was required before secure effects were observed. Increasing the intensity of indentation (resulting in activation of greater numbers of joint receptors) increases the strength of coupling between joint afferent input and motor unit responses, although the relationship is not linear. The relation between the discharge patterns of pairs of motor units was also examined, and it was found that significant coupling occurred at the stimulus frequency, superimposed on a 'background' coupling from unidentified sources. The phase relationship between pairs of motor units was not affected by the periodic stimulus. However, it was observed that if two motor units were firing independently of one another in the absence of capsule probing, maintained capsular indentation produced a striking synchronization between the discharges of the two motor units. These experiments show strong joint-mediated reflex effects on motor unit firing indicating that joint receptors may have an important role to play in motor control.

Perceptual responses to microstimulation of single afferents innervating joints, muscles and skin of the human hand.

1. Microneurographic techniques were used to isolate single afferent axons within cutaneous and motor fascicles of the median and ulnar nerves at the wrist in thirteen subjects. Of the sixty-five identified afferents, eleven innervated the interphalangeal and metacarpophalangeal joints, sixteen innervated muscle spindles, three innervated Golgi tendon organs and thirty-five supplied the glabrous skin of the hand. 2. Intrafascicular stimulation through the recording microelectrode, using trains of constant-voltage positive pulses (0.3-0.8 V, 0.1-0.2 ms, 1-100 Hz) or constant-current biphasic pulses (0.4-13.0 microA, 0.2 ms, 1-100 Hz), evoked specific sensations from sites associated with some afferent species but not others. 3. Microstimulation of eight of the eleven joint afferent sites (73%) evoked specific sensations. With four, subjects reported innocuous deep sensations referred to the relevant joint. With the other four, the subjects reported a sensation of joint displacement that partially reflected the responsiveness of the afferents to joint rotation. 4. Microstimulation of fourteen of the sixteen muscle spindle afferent sites (88%) generated no perceptions when the stimuli did not produce overt movement. However, subjects could correctly detect the slight movements generated when the stimuli excited the motor axons to the parent muscle. 5. With seven of the nine rapidly adapting (type RA or FAI) cutaneous afferents (88%) microstimulation evoked sensations of 'flutter-vibration', and with two of eight slowly adapting (type SAI) afferents (25%) it evoked sensations of 'sustained pressure'. Of the eighteen SAII afferents, which were classified as such by their responses to planar skin stretch, the majority (83%) generated no perceptions, confirming previous work, but three evoked sensations of movements or pressure. 6. The present results suggest a relatively secure transmission of joint afferent traffic to perceptual levels, and it is concluded that the human brain may be able to synthesize meaningful information on joint displacement on the basis of impulses in a single joint afferent. This could partly compensate for the low responsiveness of individual joint afferents within the physiological range of joint displacements. Although single muscle spindle afferents can adequately encode joint position and movement, the results suggest that the brain needs the information from more than one muscle spindle afferent to perceive changes in joint angle.

Reflex actions of knee joint afferents during contraction of the human quadriceps

(1) The spinal reflex actions of afferents stimulated by knee joint distension have been investigated in man. (2) Cannulation of the knee and infusion of saline raised intra-articular pressure, especially during quadriceps contraction. High pressures did not induce any sensation of pain. Pressure was taken as an index of joint proprioceptor activation. (3) Increased pressure progressively depressed the quadriceps H-reflex, both at rest and during quadriceps contraction. There was no indication of a threshold pressure for this inhibitory action. (4) It is concluded that joint distension inhibits quadriceps motoneurons through spinal pathways that still operate during voluntary contraction. These pathways could thus contribute to pathological weakness after joint injury. (5) Joint distension produced spatial facilitation of non-reciprocal inhibition of quadriceps H-reflexes from afferents in the tibial nerve.

A group II-activated ascending tract of lumbosacral origin in the cat spinal cord.

1. Electrophysiological investigations have revealed a population of ascending tract neurones originating in the lumbosacral enlargement, with input from group II muscle afferents of the cat hindlimb. 2. Single-unit microelectrode recordings were made in the lateral funiculus at L6, from the axons of thirty-four ascending tract neurones. All of the axons were antidromically activated by stimulation of the ipsilateral lateral funiculus at Th13 and, whenever tested (eight units), at C1. 3. Conduction velocities of the axons, between the L6 and Th13 segment, ranged from 33 to 92 m s-1 (mean 61 m s-1). 4. All of the ascending tract neurones were discharged following electrical stimulation of muscle nerves at group II strength, but not by weaker stimuli in the group I range. Most of the investigated neurones were excited by group II afferents of more than one muscle nerve. In addition, a proportion of the units tested could also be discharged by cutaneous and by joint afferents. 5. Responses to natural stimuli were investigated in eighteen ascending tract neurones discharged by electrical stimulation of group II afferents in the gastrocnemius-soleus (GS) and plantaris (P1) nerves which were dissected free in continuity with their muscles. Seven units were spontaneously active. Eight units responded to isometric contraction of the GS/P1 muscles with a discharge occurring mainly on the falling phase of muscle tension. Nine units increased their discharge frequency in response to stretching of the muscles and five units responded to mechanically probing the muscles with a blunt instrument. 6. The final termination sites of this group of ascending tract neurones has yet to be determined. Initial attempts (three units) to antidromically activate the neurones from the cerebellum have been unsuccessful. Other likely areas of termination in the brain stem are considered.

Joint Innervation in Joint Injury

When considering joint innervation, it is important to appreciate that the joint receptors and the muscle tendon units about the joint interact in a complimentary manner forming a single afferent system. This system remains complex, and its overall function is still relatively unclear. Although the role of joint afferents can not be explicitly described, it is important to appreciate that alterations in joint innervation caused by athletic trauma can occur and can markedly affect joint function.

The cortical distribution of muscle and cutaneous afferent projections from the human foot

Somatosensory evoked potentials to electrical stimulation of muscle and cutaneous afferents from the foot were recorded in normal human subjects using multiple channels centered on the vertex and referenced to the contralateral earlobe. Low-threshold muscle afferents were selectively activated by an insulated microelectrode inserted percutaneously at the motor point of abductor hallucis. Low-threshold cutaneous and joint afferents of the hallux or second toe were stimulated with ring electrodes. The posterior tibial and sural nerves were stimulated at the ankle through surface electrodes. The cerebral distribution of the initial cortical responses (N33-P40) to stimulation of muscle afferents largely paralleled that to stimulation of its parent nerve, the posterior tibial nerve (which contains afferents of muscle, cutaneous and joint origin). They were maximal slightly posterior and ipsilateral to the vertex. The cutaneous-joint afferent projection from the hallux paralleled that from the sural nerve and both were less lateralized than the tibial and abductor hallucis projections.

Group II-activated lumbosacral interneurones with an ascending projection to midlumbar segments of the cat spinal cord.

1. In anaesthetized cats, single-unit microelectrode recordings were made in the lateral funiculus at L6, from the axons of lumbosacral interneurones discharged by hindlimb group II muscle afferents. 2. The level of the ascending projection of these interneurones was investigated by antidromic activation of their axons in the lateral funiculus from different spinal levels. The majority of units encountered were found to have an ascending projection to at least the L4 level and, of these, most (85%) did not project beyond the L4 or L3 segments of the cord. 3. The axons studied were discharged by group II afferents primarily from knee extensor muscles. Some units were discharged in addition by cutaneous and/or joint afferents. 4. The implications of this ascending projection are discussed.

Tonic suppression of reflex transmission in low spinal cats.

Transmission in various spinal hindlimb reflex pathways arising from muscle, cutaneous and joint afferents, was investigated in acute low spinal (Th10) cats before and after lesions at more caudal levels in the lumbar segments. Lesions of the ipsilateral dorsolateral funiculus (DLF) resulted in increased mono- and polysynaptic ventral root discharges. With conditioning of monosynaptic reflexes it was demonstrated that the DLF lesion enhanced transmission in reflex pathways from group II and III muscle afferents. The DLF lesion also resulted in increased dorsal root potentials from cutaneous, joint and group III muscle afferents. These reflex enhancements could not be obtained with lesions rostral to L2, but developed with lesions in L2 and to some extent in L3, but no further effect was obtained by adding lesions caudal to L3. Ventral extension of the DLF lesion gave hardly any increase of reflex transmission. It is postulated that the investigated reflex pathways may be tonically inhibited in the acute low spinal state by propriospinal neurones with cell bodies in the L2-L3 segments and with axons descending in the dorsolateral funiculus.

Differences in the chemical expression of rat primary afferent neurons which innervate skin, muscle or joint

The fluorescent dye Fast Blue was injected in anaesthetized rats into either skin, muscle or knee joint of the hindlimb. Following retrograde transport of the dye to lumbar dorsal root ganglia, the cell bodies of primary afferent neurons innervating these different target tissues were identified in ganglion sections by fluorescence microscopy. The sections were processed to demonstrate activity of the enzyme thiamine monophosphatase, or immunoreactivity to calcitonin gene-related peptide, substance P, or somatostatin, in Fast Blue labelled neurons. In all cases immunoreactivity to the antineurofilament antibody RT97 was used to classify dorsal root ganglion cells as being either small dark (RT97 negative, unmyelinated axons) or large light (RT97 positive, myelinated axons). The proportion of small dark cells labelled from each target decreased in the order: skin, muscle, joint. Thiamine monophosphatase and somatostatin were present only in small dark cells, while calcitonin gene-related peptide and substance P were found in both small dark and large light cells. In large light cells of all three targets, more contained calcitonin gene-related peptide than substance P. Among small dark cells, thiamine monophosphatase and somatostatin were found predominantly in skin afferents, while calcitonin gene-related peptide and substance P were more common in muscle and joint afferents. The chemical expression of primary afferents is therefore characteristic of the peripheral target they innervate. This could reflect either a maintained influence of the target on the afferents, or the factors which operate only during development.