Effect of ageing on tactile transduction processes

With advancing age, a decline in the main sensory modalities including touch sensation and perception is well reported to occur. This review mainly outlines the peripheral components of touch perception highlighting ageing influences on morphological and functional features of cutaneous mechanical transducers and mechanosensitive ion channels, sensory innervation, neurotransmitters and even vascular system required to ensure efferent function of the afferent nerve fibres in the skin. This, in conjunction with effect of ageing on the skin per se and central nervous system, could explain the tactile deficit seen among the ageing population. We also discuss appropriate tools and experimental models available to study the age-related tactile decline.     

Sensory nerve endings in the anterior cruciate ligament (Lig. cruciatum anterius) of sheep

This study examines the structure of sensory nerve endings in the sheep anterior cruciate ligament (ACL). Three types of nerve endings are found: free nerve endings (FNE), Ruffini corpuscles, and lamellated corpuscles. The FNE (more than 100) are found subsynovially. The afferent nerve fibres are either thin myelinated axons (Aδ) or C fibres with diameters of 1–2 μm. FNE have been reported to function as thermoreceptors and polymodal nociceptors. In addition, FNE are also seen between fascicles of collagen fibres, often close to blood vessels. Part of this group may be efferent autonomic fibres controlling local blood flow. The corpuscles are seen subsynovially and between fascicles of connective tissue close to the attachment points of the ACL. A ligament contains about 20 Ruffini corpuscles, which are mainly located in the subsynovial connective tissue. They consist of cylinders formed from perineural cells surrounding the afferent myelinated axons (diameters 4–5 μm) with enlarged nerve terminals anchored between collagen fibres. These enter in bundles from the surrounding connective tissue at one open pole, pass through the length of the cylinder, and leave at the other pole. Functionally, Ruffini corpuscles have been described as slowly adapting stretch receptors. Lamellated corpuscles (usually between 5 and 15) are found in the subsynovial connective tissue. The afferent myelinated axon has a diameter of 4–6 μm, and the nerve terminal is located in the centre of numerous layers formed by lamellated terminal glial cells and by a perineural capsule. They are known to function as rapidly adapting pressure receptors. The most important function of the ACL is its mechanical function, but additional sensory functions must be considered triggering reflex mechanisms in case of extreme positioning or overload. Anat Rec 254:13–21, 1999. © 1999 Wiley‐Liss, Inc.     

Myelin basic protein-positive nerve fibres in human Meissner corpuscles

Myelinated nerve fibres forming sensory corpuscles become amyelinic before entering the corpuscle. Interestingly, in Meissner corpuscles from monkey myelin basic protein (MBP), a specific component of myelin sheath co-localized with neuronal markers. To investigate whether or not this also occurs in human digital Meissner corpuscles, we used single and double immunohistochemistry to detect MBP associated with axonic (protein gene product (PGP) 9.5) or Schwann and Schwann-related cell (S100 protein) markers. We also studied these markers in Pacinian corpuscles. Nerve fibres immunoreactive for MBP were detected in about 25% of the Meissner corpuscles examined; however, MBP never co-localized with PGP 9.5 and MBP occasionally co-localized with S100 protein. MBP-immunoreactive fibres associated with Meissner corpuscles were observed at the periphery of the lamellar cells or within the corpuscle between the lamellar cells. These results describe the distribution of myelinated nerve fibres expressing MBP in human Meissner corpuscles, which is important when studying Meissner corpuscles in cutaneous biopsies used for the diagnosis of peripheral and degenerative neuropathies.     

Synaptic transmission between single slowly adapting type I fibres and their cuneate target neurones in cat.

1. The synaptic linkage between single, identified slowly adapting type I (SAI) fibres and their central target neurones of the cuneate nucleus was examined in pentobarbitone-anaesthetized cats. Simultaneous extracellular recordings were made from individual cuneate neurones and from fine, intact fascicles of the lateral branch of the superficial radial nerve in which it was possible to identify and monitor the activity of each group II fibre. Individual SAI fibres were activated by static displacement and by vibration delivered with a fine probe (0.25-2 mm diameter) to their associated touch domes in the hairy skin of the forelimb. 2. Transmission properties across the synapse were analysed for nine SAI-cuneate pairs in which the single SAI fibre of each pair provided a suprathreshold input to the cuneate neurone. Neither spatial nor temporal summation was required for effective impulse transmission, and often more than 80% of SAI impulses led to a response in the cuneate neurone. Responses of the cuneate neurones to single SAI impulses occurred at a short, fixed latency (S.D. often < 0.1 ms), and frequently consisted of a burst of two or three impulses, at low SAI input rates in particular. 3. The tight phase-locking in the responses to vibration of single SAI fibres was preserved in the cuneate responses for frequencies up to approximately 400 Hz. However, as the impulse rates of the cuneate neurones were less than 150 impulses s-1, their impulse patterns could not directly signal the vibration periodicity at frequencies > 100-150 Hz despite 1:1 responses in their single SAI input fibres up to approximately 500 Hz. 4. The reliable transmission of touch dome-associated SAI input across the cuneate nucleus indicates that transmission failure at this first relay is unlikely to be responsible for the reported failure of touch dome-SAI inputs to contribute to tactile perception. 5. The transmission characteristics for the SAI fibres were very similar to those demonstrated previously for fibres associated with Pacinian corpuscles, which argues against any marked differential specialization in transmission characteristics for dorsal column nuclei neurones that receive input from different tactile fibre classes.     

Wagner-Meissner neurilemmoma of the right cheek

We report a very rare case of Wagner-Meissner neurilemmoma in the cheek of a 10-year-old boy. The tumor presented as a slowly growing soft tissue swelling. Magnetic resonance imaging disclosed a very infiltrative, 9-cm mass involving the subcutis and deep soft tissues of the right cheek. Microscopically, the tumor was unencapsulated and composed almost entirely of well-formed Wagner-Meissner corpuscles that formed confluent sheets, perivascular cuffs, and individual corpuscles percolating through adipose tissue. Compared with the 3 previous reports, which describe circumscribed, encapsulated tumors in adult patients, this case had distinctive clinicopathologic features never reported: presentation in a pediatric patient, location in the head and neck region, and an infiltrative growth pattern.     

Further observations on the sensitive innervation of some bird's proctodeum

The AA. studied the autonomic and sensitive somatic innervation of some female bird's proctodeum, through the properly modified Ruffini's gold chloride method. The vegetative component was constituted by ganglion cells of different size, isolated or grouped to form ganglia, found along the course of nerve trunks or in the concurrent point of different nerve bundles. The sensitive somatic innervation was represented by free and encapsulated endings differently distributed in the thickness of the wall. The former were composed of thin networks, while the latter, located more frequently in the muscular tunica and in the subadventitial connective, were composed of encapsulated receptors classified as Pacini, Pacini-like and Herbst corpuscles. The morphology of these receptors was described and hypotheses were brought up about their probable functional role. The AA, also found, even if very rarely, helicoidal collagen fibres around nerve fascicles.     

Merkel cells and Meissner's corpuscles in human digital skin display Piezo2 immunoreactivity

The transformation of mechanical energy into electrical signals is the first step in mechanotransduction in the peripheral sensory nervous system and relies on the presence of mechanically gated ion channels within specialized sensory organs called mechanoreceptors. Piezo2 is a vertebrate stretch‐gated ion channel necessary for mechanosensitive channels in mammalian cells. Functionally, it is related to light touch, which has been detected in murine cutaneous Merkel cell–neurite complexes, Meissner‐like corpuscles and lanceolate nerve endings. To the best of our knowledge, the occurrence of Piezo2 in human cutaneous mechanoreceptors has never been investigated. Here, we used simple and double immunohistochemistry to investigate the occurrence of Piezo2 in human digital glabrous skin. Piezo2 immunoreactivity was detected in approximately 80% of morphologically and immunohistochemically characterized (cytokeratin 20⁺, chromogranin A⁺ and synaptophisin⁺) Merkel cells. Most of them were in close contact with Piezo2⁻ nerve fibre profiles. Moreover, the axon, but not the lamellar cells, of Meissner's corpuscles was also Piezo2⁺, but other mechanoreceptors, i.e. Pacinian or Ruffini's corpuscles, were devoid of immunoreactivity. Piezo2 was also observed in non‐nervous tissue, especially the basal keratinocytes, endothelial cells and sweat glands. The present results demonstrate the occurrence of Piezo2 in cutaneous sensory nerve formations that functionally work as slowly adapting (Merkel cells) and rapidly adapting (Meissner's corpuscles) low‐threshold mechanoreceptors and are related to fine and discriminative touch but not to vibration or hard touch. These data offer additional insight into the molecular basis of mechanosensing in humans.     

Nerve fibres are required to evoke a contact sensitivity response in mice

Previous work has indicated that the dermis and epidermis of skin contains abundant nerve fibres closely associated with Langerhans' cells. We have investigated whether these nerve endings are necessary for inducing and evoking a contact sensitivity (CS) response. Topical application of a general or a peptide (calcitonin gene-related peptide and substance P)-specific neurotoxin was employed to destroy the nerve fibres at skin sites subsequently used to induce or evoke the CS response. Elimination of nerve fibres abolished both induction and effector stages of the specific CS response. Denervation did not destroy the local Langerhans' cells, which were observed in increased numbers, or prevent them from migrating to lymph nodes. The local CS response was also abolished by systemic deletion of capsaicin-sensitive nerve fibres, suggesting that the loss of response was not non-specific but associated with the loss of specific nerve fibres. The results indicate that peptidergic nerve fibres are required to elicit a CS response and may be vital to the normal function of the immune system.     

Ageing of the somatosensory system at the periphery: age‐related changes in cutaneous mechanoreceptors

Decline of tactile sensation associated with ageing depends on modifications in skin and both central and peripheral nervous systems. At present, age‐related changes in the periphery of the somatosensory system, particularly concerning the effects on mechanoreceptors, remain unknown. Here we used immunohistochemistry to analyse the age‐dependent changes in Meissner's and Pacinian corpuscles as well as in Merkel cell‐neurite complexes. Moreover, variations in the neurotrophic TrkB‐BDNF system and the mechanoprotein Piezo2 (involved in maintenance of cutaneous mechanoreceptors and light touch, respectively) were evaluated. The number of Meissner's corpuscles and Merkel cells decreased progressively with ageing. Meissner's corpuscles were smaller, rounded in morphology and located deeper in the dermis, and signs of corpuscular denervation were found in the oldest subjects. Pacinian corpuscles generally showed no relevant age‐related alterations. Reduced expression of Piezo2 in the axon of Meissner's corpuscles and in Merkel cells was observed in old subjects, as well was a decline in the BDNF‐TrkB neurotrophic system. This study demonstrates that cutaneous Meissner's corpuscles and Merkel cell‐neurite complexes (and less evidently Pacinian corpuscles) undergo morphological and size changes during the ageing process, as well as a reduction in terms of density. Furthermore, the mechanoprotein Piezo2 and the neurotrophic TrkB‐BDNF system are reduced in aged corpuscles. Taken together, these alterations might explain part of the impairment of the somatosensory system associated with ageing.     

Sensory Innervation of the Female Human Umbilical Skin: Morphological Studies

Background: Sensory stimuli are conducted by several cutaneous sensory nerves and tactile corpuscles. The latter are specialized sensory organs that represent the starting point of many afferent sensory pathways. To date, our knowledge about the distribution of the sensory innervation in the umbilical skin of females is incomplete.

Aim of the study: To elucidate the morphology of the cutaneous innervation of the normal female umbilical skin.

Materials and methods: Biopsies of normal umbilical skin were obtained from female patients undergoing umbilical hernial repair. The specimens were processed for both immunohistological (antibodies against PGP9.5, pan-neuronal marker, and S-100 protein, marker of Schwann cells) and ultrastructural (transmission electron microscopy) examinations.

Results: The authors found abundant genital end-bulb-like structures, numerous epidermal and dermal Merkel cells, Meissner and Ruffini corpuscles, intraepidermal nerve terminals, and multiple free nerve endings surrounding the ducts and acini of the sweat glands.

Conclusions: The umbilical skin of females has abundant sensory innervation similar to that of the glans penis.     

Functional role of unmyelinated tactile afferents in human hairy skin: sympathetic response and perceptual localization

In addition to A-beta fibres the human hairy skin has unmyelinated (C) fibres responsive to light touch. Previous functional magnetic resonance imaging (fMRI) studies in a subject with a neuronopathy who specifically lacks A-beta afferents indicated that tactile C afferents (CT) activate insular cortex, whereas no response was seen in somatosensory areas 1 and 2. Psychophysical tests suggested that CT afferents give rise to an inconsistent perception of weak and pleasant touch. By examining two neuronopathy subjects as well as control subjects we have now demonstrated that CT stimulation can elicit a sympathetic skin response. Further, the neuronopathy subjects’ ability to localize stimuli which activate CT afferents was very poor but above chance level. The findings support the interpretation that the CT system is well suited to underpin affective rather than discriminative functions of tactile sensations.     

Immunohistochemical demonstration of neuron specific enolase (NSE) on cutaneous nerves: comparative study using NSE and S-100 protein antibodies on denervated skin

The presence of neuron-specific enolase (NSE) and S-100 protein, which are nerve specific proteins, was immunohistochemically investigated on the cutaneous nerves. NSE was found in the axons of the cutaneous nerve bundles and the terminal axons in the Meissner and Pacinian corpuscles of the normal human and macaque skin. S-100 protein was found in the Schwann cells, lamellar cells of the Meissner corpuscles, and inner core cells of the Pacinian corpuscles. After denervation of the ulnar nerve on macaque, NSE on axons of the cutaneous nerves and Meissner and Pacinian corpuscles was completely disappeared in the 5th digit. However, S-100 protein was still maintained in the Schwann cells and Meissner and Pacinian corpuscles in the same digit. From these results, we conclude that the comparative immunohistochemical staining of NSE and S-100 protein is simple and reliable method to demonstrate the cutaneous nerves in normal and pathological conditions.     

Clustering of Pacinian corpuscle afferent fibres in the human median nerve

 To further study the functional organisation of human peripheral nerves, the intrafascicular arrangement of afferent fibres supplying Pacinian corpuscles (PCs) was explored by percutaneous microneurography using thin-calibre, concentric needle electrodes. In normal adults, 20 PC afferents were identified in 13 recording sites. Low-amplitude (less than 30 μm) vibratory stimuli to the skin were applied with tuning forks oscillating at 128 Hz or 256 Hz and response patterns of individual PC units were studied. In many recording sites, two, sometimes even three, PC afferents with adjacent or overlapping receptive fields in the hand were clustered in the nerve. The observed incidence in the records containing a certain number of PC units was compared with the expected probability calculated according to the hypothesis that all nerve fibres are randomly organised in peripheral nerves. The results suggested that PC afferents are partially segregated in the nerve. In addition, PC afferents were neighbouring on slowly adapting type II (SAII) units and skin sympathetic activity in individual fascicles. SAII units often innervated the same skin area as PC units, but did not respond to vibration. The data provided additional information regarding the functional organisation of the human peripheral nerve and the mechanisms underlying the sense of vibration in man with special regard to population behaviour of neighbouring PC mechanoreceptors. Received: 25 September 1998 / Accepted: 14 December 1998     

Immunohistological demonstration of S-100 protein in the cutaneous nervous system

S-100 protein, isolated from mammalian brain, has widely been used for immunohistochemical marker of the glia cells and the cells derived from the neural crest. In the present study, we made anti S-100 protein antibody and studied the immunoreactive distribution of S-100 protein in the cutaneous nervous system. Albino rabbits were immunized with S-100 protein and complete Freund adjuvant, and antiserum was purified by ion-exchange chromatography. Formalin-fixed normal human skin and sciatic nerve of rat were examined by the PAP method. S-100 protein was detected in Schwann cells of sciatic nerve of rat and cutaneous nerve bundles of human skin specimens. Meissner corpuscles and inner core cells of Pacinian corpuscles of human skin were S-100 protein positive. These findings suggest that the staining of S-100 protein with PAP method is a simple and reliable method to demonstrate the cutaneous nervous system. Also, lamellar cells of Meissner corpuscles and inner core cells of Pacinian corpuscles are indicated to be Schwann cell origin.     

Skin biopsy for the diagnosis of peripheral neuropathy

Skin biopsy has become an accepted tool for investigating small nerve fibres, which are invisible to conventional neurophysiological tests even though they are affected early on in peripheral neuropathies of varying aetiology. Morphometric analysis of epidermal and dermal nerves has proved to be reliable, reproducible and unaffected by the severity of neuropathy, making skin biopsy useful for diagnosing small fibre neuropathy (SFN) in clinical practice. The possibility of obtaining skin biopsy specimens from different sites of the body, to repeat them within the area of the same sensory nerve, to distinguish between somatic and autonomic nerves and to investigate the expression of nerve-related proteins has widened the potential applications of this technique to clinical research. Skin biopsy performed using a minimally invasive disposable punch is a safe and painless procedure. Using specific antibodies with bright-field immunohistochemistry or immunofluorescence technique, it is possible to investigate unmyelinated fibres innervating the epidermis of hairy and glabrous skin, large myelinated fibres supplying specialized corpuscles in glabrous skin, and autonomic fibres innervating sweat glands, blood vessels and arrector pilorum muscles. This review discusses the features of skin innervation in hairy and glabrous skin, the functional properties of skin nerve fibres and their changes in peripheral neuropathies.     

The capsule of human Meissner corpuscles: immunohistochemical evidence

Meissner corpuscles are cutaneous mechanoreceptors that are usually located in the dermal papillae of human glabrous skin. Structurally, these sensory corpuscles consist of a mechanoreceptive sensory neuron surrounded by non-myelinating lamellar Schwann-like cells. Some authors have described a partially developed fibroblastic capsule of endoneurial or perineurial origin around Meissner corpuscles; however, others have noted that these structures are non-encapsulated. As there is continuity between the periaxonic cells forming the sensory corpuscles and the cells of the nerve trunks, we used immunohistochemistry to examine the expression of endoneurial (CD34 antigen) or perineurial [Glucose transporter 1 (Glut1)] markers in human cutaneous Meissner corpuscles. We also investigated the immunohistochemical patterns of nestin and vimentin (the main intermediate filaments of the cytoskeleton of endoneurial and perineurial cells, respectively) in Meissner corpuscles. The most important finding from this study was that CD34-positive cells formed a partial/complete capsule of endoneurial origin around most Meissner corpuscles, without signs of other perineurial Glut1-positive elements. However, the cytoskeletal proteins of the capsular CD34-positive cells did not include either nestin or vimentin, so the cytoskeletal composition of these cells remains to be established. Finally, the intensity of the immunoreactivity for CD34 in the capsule decreased with ageing, sometimes becoming completely absent in the oldest individuals. In conclusion, we report the first immunohistochemical evidence of the capsule of Meissner corpuscles in humans and demonstrate the endoneurial origin of the capsule.     

Immunohistochemical localization of laminin and type IV collagen in human cutaneous sensory nerve formations

We used immunohistochemical techniques and monoclonal antibodies to localize two basement membrane components (laminin and type IV collagen) in the nerves and sensory nerve formations, or corpuscles, supplying human digital skin. Furthermore, neurofilament proteins, S-100 protein and epithelial membrane antigen were studied in parallel. In dermal nerve trunks, immunostaining for laminin and type IV collagen was found to be co-localized in the perineurium and the Schwann cells, the stronger immunoreactivity being at the external surface of the cells. In the Meissner digital corpuscles, the immunoreactivity for laminin and type IV collagen was mainly observed underlying the cell surface of lamellar cells, while the cytoplasm was weakly immunolabelled or unlabelled. Finally, within Pacinian corpuscles co-localization of the two basement membrane molecules was encountered in the inner core, intermediate layer, outer core and capsule. Laminin and type IV collagen immunoreactivities were also found in blood vessels and sweat glands, apparently labelling basement membrane structures. The present results provide evidence for the presence of basement membrane in all periaxonic cells forming human cutaneous sensory nerve formations, and suggest that all of them are able to synthesize and release some basement membrane components, such as laminin and type IV collagen. The possible role of laminin in sensory nerve formations is discussed.     

Digital pacinian neuroma: a distinctive hyperplastic lesion

Neural tumours composed solely of Pacinian corpuscles or showing focal Pacinian differentiation are extremely rare and have only occasionally been reported in the literature. All such lesions to date have been benign. Three lesions are described herein which presented as painful digital masses in middle-aged adults and which were composed of abnormal aggregates of morphologically mature Pacinian corpuscles and intervening small nerves. Only five similar cases have been previously recorded. Possible pathogenetic mechanisms of this unusual hyperplastic phenomenon are discussed.     

p75 and TrkA neurotrophin receptors in human skin after spinal cord and peripheral nerve injury,with special reference to sensory corpuscles

Human skin, including nerves and sensory corpuscles, displays immunoreactivity (IR) for low- (p75) and high-affinity (TrkA-like) receptors for nerve growth factor (NGF), the best characterized member of the family of neurotrophins. This study was designed to analyze the changes induced by spinal cord and peripheral nerve injuries in the expression of neurotrophin receptors in digital skin, with special reference to nerves and sensory corpuscles. Skin biopsy samples were obtained from 1) the hand and toes of normal subjects, 2) below the level of the lesion of patients with spinal cord injury affecting dorsal and lateral funiculi, 3) the cutaneous territory of entrapped peripheral nerves (median and ulnar nerves), and 4) the cutaneous territory of sectioned and grafted nerves (median nerve). The pieces were formalin-fixed and paraffin-embedded, cut in serial sections, and processed for immunohistochemistry using antibodies against human p75 and TrkA proteins. The percentage of sensory corpuscles displaying IR for p75 and TrkA-like, as well as the intensity of IR developed within them, was assessed using quantitative image analysis. Spinal cord severance causes a decrease in p75 IR in Meissner and Pacinian corpuscles, whereas TrkA-like IR did not vary. In other nonnervous tissues (i.e., epidermis, sweat glands), both p75 and TrkA-like IR was diminished or even absent. Similar but more severe changes were encountered in the skin from the territory of entrapped nerves. Finally, in subjects with sectioned-grafted nerves, p75 IR was found close to controls in nerves, reduced in Meissner corpuscles, and absent in the inner core of the Pacinian ones; TrkA-like IR was in the perineurium, a small percentage of Meissner corpuscles (about 7%), and the outer core and capsule of the Pacinan corpuscles. In the nonnervous tissues, p75 IR was practically absent, whereas TrkA-like IR did not change. No changes in the expression of neurotrophin receptors were observed in Merkel cells of the different groups. Present results show the following: 1) expression of nerve p75 IR in human cutaneous sensory corpuscles is sensitive to central deafferentation, to blockade or difficulty in axonal transport, and to disruption of axonalcontinuity independently of possible restoration of axonal integrity due to grafts; 2) expression of TrkA-like IR in nerves and sensory corpuscles is sensitive only to nerve transection; 3) the corpuscular Schwann-related cells are the only cells involved in the above modifications, the perineurial cells remaining unchanged; 4) the expression of p75 and TrkA-like IR by Merkel cells is independent of normal innervation; 5) an adequate innervation of the skin seems to be necessary for the expression of p75 but not TrkA-like in nonneuronal cells, especially in the epidermis. A role for NGF in the maintenance of epidermis integrity is discussed. Anat. Rec. 251:371–383, 1998. © 1998 Wiley-Liss, Inc.