Morphological Evidence for the Sensitivity of the Ear Canal of Odontocetes as shown by Immunohistochemistry and Transmission Electron Microscopy.

The function of the external ear canal in cetaceans is still under debate and its morphology is largely unknown. Immunohistochemical (IHC) analyses using antibodies specific for nervous tissue (anti-S100, anti-NSE, anti-NF, and anti-PGP 9.5), together with transmission electron microscopy (TEM) and various histological techniques, were carried out to investigate the peripheral nervous system of the ear canals of several species of toothed whales and terrestrial Cetartiodactyla. This study highlights the innervation of the ear canal with the presence of lamellar corpuscles over its entire course, and their absence in all studied terrestrial mammals. Each corpuscle consisted of a central axon, surrounded by lamellae of Schwann receptor cells, surrounded by a thin cellular layer, as shown by IHC and TEM. These findings indicate that the corpuscles are mechanoreceptors that resemble the inner core of Pacinian corpuscles without capsule or outer core, and were labelled as simple lamellar corpuscles. They form part of a sensory system that may represent a unique phylogenetic feature of cetaceans, and an evolutionary adaptation to life in the marine environment. Although the exact function of the ear canal is not fully clear, we provide essential knowledge and a preliminary hypothetical deviation on its function as a unique sensory organ.

Immunohistochemical profile of human pancreatic pacinian corpuscles.

OBJECTIVES: To analyze the immunohistochemical profile of the human pancreatic pacinian corpuscles in comparison with that of the cutaneous pacinian corpuscles. In addition, we studied a Pacinilike corpuscle found in the adventitia of a pancreatic artery. METHODS: We used immunohistochemistry to detect specific antigens for corpuscular constituents, specific antibodies for the identification of Adelta- and C-sensory fibers and for the detection of several growth factor receptors, and some members of the degenerin/epithelial Na channel superfamily of proteins. RESULTS: Approximately 62% of pancreatic pacinian corpuscles have 2 to 10 axonic profiles each enclosed by its own inner core: 1 or 2 of these axonic profiles displayed RT-97 immunoreactivity (specific marker of mechanical axons). The cutaneous pacinian corpuscles showed not more than 2 axonic profiles with identical immunohistochemical characteristics. The expression of glial fibrillary acidic protein, epithelial membrane antigen, and tyrosine receptor kinase B was different between pancreatic and cutaneous pacinian corpuscles; the pattern of distribution of degenerin/epithelial Na channel proteins was identical in both cases. The arterial Pacinilike corpuscles displayed a specific immunohistochemical profile. CONCLUSIONS: Pancreatic pacinian corpuscles slightly differ from the cutaneous ones, and these differences could be related to topography, growth factor requirements, or function of pacinian corpuscles in the pancreas.

A small lamellar corpuscle within a small nerve bundle outside the tendon of the rat soleus muscle.

A small nerve bundle outside the tendon of the adult rat soleus muscle contained a small lamellar corpuscle similar in structural organization to the ordinary paciniform corpuscle. A terminal axon composing this corpuscle was originated from a side branch of an afferent nerve fiber and surrounded by a number (approximately 15) of closely packed flattened lamellae of modified Schwann cells, while the stem nerve fiber freely terminated within the nerve bundle. These findings suggested that an afferent nerve fiber retracted after degeneration might extend a new branch within the nerve bundle and unexpectedly form a lamellar corpuscle within it.

Possible glutaminergic interaction between the capsule and neurite of Pacinian corpuscles.

The role of the capsule encasing the Pacinian corpuscle's (PC's) neurite, where mechanotransduction occurs, may be more than mechanical. The inner core of the PC's capsule consists of lamellar cells that are of Schwann-cell origin. Previously, we found both voltage-gated Na+ and K+ channels in these inner-core lamellae. Research on astrocytes and Schwann cells shows bidirectional signaling between glia and neurons, a major component of which is glutamate. Furthermore, Merkel cells show positive immunoreactivity for glutamate receptor mGluR5, and the glutamate-receptor antagonist kynurenate greatly decreases the static activity of the slowly adapting neurons of Merkel cell-neurite complexes. To investigate the possibility of glutaminergic interaction in PCs, we applied antibodies to glutamate, glutamate receptors, glutamate transporters, and SNARE proteins to cat mesenteric PC sections. Positive labeling was seen in the inner-core lamellae, at inter-lamellar connections, where the lamellae contact the membrane of the neurite and at the lamellar tips. The presence of these proteins on the lamellae and neurite membranes, demonstrated both with immunofluorescent light microscopy as well as immunogold electron microscopy, suggests a chemical, possibly bidirectional, interaction between the lamellar cells and the neurite. Thus, the capsule of the PC, apart from having a mechanical filtering function, may also provide an environment for lamellar-neurite interaction, perhaps acting as a neuro-modulator of the initiation, and/or continuation, of the mechanical-electrical transduction process. At the very least, the presence of the aforementioned proteins suggest some sort of "synaptic-like" activity in these mechanoreceptors, which up until now has not been considered possible.

Anatomy of the deep fascia of the upper limb. Second part: study of innervation.

Analysis of specimens taken from different areas of the deep fascia in 20 upper limbs was made in order to establish which kind of nerve fibres and endings are present in the deep muscular fascia. The flexor retinaculum and the lacertus fibrosus were also evaluated because they are anatomically hardly separable from the deep muscular fascia, although they have different functions. In particular, specimens were taken at the level of: (a) the expansion of pectoralis major onto the bicipital fascia, (b) the middle third of the brachial fascia, (c) the lacertus fibrosus, (d) the middle third of the antebrachial fascia, (e) the flexor retinaculum. This study demonstrated an abundant innervation of the fascia consisting in both free nerve endings and encapsulated receptors, in particular, Ruffini and Pacini corpuscles. However, differences in innervation were verified: the flexor retinaculum was resulted the more innervated element whilst lacertus fibrosus and the pectoralis major expansion the less innervated. These results suggest that the retinaculum has more a perceptive function whereas the tendinous expansions onto the fascia have mostly a mechanical role in the transmission of tension. The hypothesis that the fascia plays an important role in proprioception, especially dynamic proprioception, is therefore advanced. In fact, the fascia is a membrane that extends throughout the whole body and numerous muscular expansions maintain it in a basal tension. During a muscular contraction these expansions could also transmit the effect of the stretch to a specific area of the fascia, stimulating the proprioceptors in that area.

Myelinated nerve endings in human skin.

We used immunohistochemical techniques and confocal microscopy to study the morphometry of myelinated nerve endings in glabrous and hairy skin. A total of 30 healthy volunteers took part in this study designed to assess the possibility of obtaining reliable information on myelinated fibers using samples of hairy skin and to determine whether differences exist between myelinated terminations from different sites. We obtained consistent information on cutaneous myelinated terminations using hairy as well as glabrous skin samples. Myelinated endings from hairy and glabrous skin differ in density and distribution. However, from a comparison of our findings with data from nerve biopsy studies, we conclude that all cutaneous myelinated terminations are thinner terminal branches of large myelinated A beta fibers, whereas cutaneous terminations of small myelinated A delta fibers lose their myelin before entering the dermis and become indistinguishable from C-fiber terminations. The classic criteria, based on fiber size, used to distinguish myelinated fiber subgroups in sensory nerves are therefore not suitable for identifying myelinated terminations in the skin.

Fine structure of Eimer's organ in the coast mole (Scapanus orarius).

Eimer's organ is a small, densely innervated sensory structure found on the glabrous rhinarium of most talpid moles. This structure consists of an epidermal papilla containing a central circular column of cells associated with intraepidermal free nerve endings, Merkel cell neurite complexes, and lamellated corpuscles. The free nerve endings within the central cell column form a ring invested in the margins of the column, surrounding 1-2 fibers that pass through the center of the column. A group of small-diameter nociceptive free nerve endings that are immunoreactive for substance P surrounds this central ring of larger-diameter free nerve endings. Transmission electron microscopy revealed a high concentration of tonofibrils in the epidermal cells of the central column, suggesting they are more rigid than the surrounding keratinocytes and may play a mechanical role in transducing stimuli to the different receptor terminals. The intraepidermal free nerve endings within the central column begin to degrade 15 microm from the base of the stratum corneum and do not appear to be active within the keratinized outer layer. The peripheral free nerve endings are structurally distinct from their counterparts in the central column and immunocytochemical double labeling with myelin basic protein and substance P indicates these afferents are unmyelinated. Merkel cell-neurite complexes and lamellated corpuscles are similar in morphology to those found in a range of other mammalian skin.

Abnormal development of pacinian corpuscles in double trkB;trkC knockout mice.

Pacinian corpuscles depend on either Aalpha or Abeta nerve fibers of the large- and intermediate-sized sensory neurons for the development and maintenance of the structural integrity. These neurons express TrkB and TrkC, two members of the family of signal transducing neurotrophin receptors, and mice lacking TrkB and TrkC lost specific neurons and the sensory corpuscles connected to them. The impact of single or double targeted mutations in trkB and trkC genes in the development of Pacinian corpuscles was investigated in 25-day-old mice using immunohistochemistry and ultrastructural techniques. Single mutations on trkB or trkC genes were without effect on the structure and S100 protein expression, and caused a slight reduction in the number of corpuscles. In mice carrying a double mutation on trkB;trkC genes most of the corpuscles were normal with a reduction of 17% in trkB-/-;trkC+/- mice, and 8% in trkB +/-;trkC -/- mice. Furthermore, a subset of the remaining Pacinian corpuscles (23% in trkB-/-;trkC+/- mice; 3% in trkB+/-;trkC-/- mice) were hypoplasic or atrophic. Present results strongly suggest that the development of a subset of murine Pacinian corpuscles is regulated by the Trk-neurotrophin system, especially TrkB, acting both at neuronal and/or peripheral level. The precise function of each member of this complex in the corpuscular morphogenesis remains to be elucidated, though.

Hypertrophy of Pacinian corpuscles in a young patient with neurofibromatosis.

Hypertrophy of Pacinian corpuscles is a condition that has been reported very infrequently. It was thought not to be connected with neurofibromatosis. A 24-year-old female with a lifelong history of neurofibromatosis type 1, who presented with left ring finger pain and decreased sensation for over 1 year is reported. Histopathology showed hypertrophy of Pacinian corpuscles. Hypertrophy of Pacinian corpuscles should be considered in the differential diagnosis of any patient with persistent pain localized to the distal palm or digits. This is the first case reported in a patient with neurofibromatosis. The finding of this lesion in a patient with neurofibromatosis is somewhat intriguing, though the relationship of these two entities is unclear.

[Branched form of the encapsulated dermal receptors in rats]. / Vetviashchaiasia forma inkapsulirovannykh retseptorov kozhi krys.

Using electron microscope the receptors found in the zone of highest receptor sensitivity in the skin fold at the base of IV finger of rat hind limb were studied. These receptors are represented by complex incapsulated corpuscles consisting of a group of small endings similar to Pacinian corpuscles, which are densely packed and surrounded by a common connective tissue capsule. Each single primary receptor is provided with an unmyelinated receptor terminal, analogue of internal bulb, and a multilayered external bulb. Primary receptor differs from Vater-Pacinian corpuscle by a significantly smaller size, form of sensitive terminal in cross-section, small number of layers forming internal and external bulbs. Complex corpuscle is the derivative of n. tibialis myelinated sensory fiber. The later is divided after the loss of myelin forming a platform of division with several (up to 5) short sprouts and separate incapsulated receptors at their ends. The complex corpuscles described apparently are dermal mechanoreceptors.

Localization and distribution of laminin in the basal lamina of certain mechanoreceptors--an immunogold study.

The applied immunogold cytochemical technique in investigating the cytologic distribution of the laminin (LAM) molecule in the capsulated Pacinian and Herbst mechanoreceptors shows the presence of LAM around most elements of the receptor structures. The LAM immunoreactivity (LAM-IR) is best expressed in the vicinity of the perineural capsule cells of both receptor types, where it is primarily concentrated around the perinuclear regions as well as the cytoplasmic lamellae. Such a localization overlaps with the already known ultrastructural localization of a basal lamina (BL) around these cells. Laminin immunoreactivity is less well expressed around the modified Schwann cells. Even in these cells, however, there is an apparent immunoreaction around the cytoplasmic lamellae regardless of the lamellar location. In both receptor types, there is no LAM-IR in the cells of the subcapsular space. Of particular significance we consider the localization of gold particles (respectively the presence of a BL) between the innermost lamellae of the modified Schwann cells and the non-myelinated part of the receptor nerve fiber and their endings, as well as around the axoplasmic protrusions of the nerve endings. We discuss the role of the BL and LAM in the investigated rapidly adapting mechanoreceptors and their trophic influence upon the sensory regions. We also assume the arresting and selective effect of these membranes in building up the ion channels of the axolemma which probably has a certain importance in mechanotransduction.

Voltage-gated sodium channels are present on both the neural and capsular structures of Pacinian corpuscles.

It has long been accepted that action potentials arising from Pacinian corpuscles (PCs) originate at the first node of Ranvier located within the PC and that the mechanotransduction events (receptor potentials) are formed by stretch-activated channels selectively sensitive predominantly to Na+. Also, it has been shown previously that tetrodotoxin (TTX) affects the receptor potential suggesting that transduction may involve voltage-sensitive Na+ channels. To determine whether voltage-sensitive Na+ channels exist in the membrane thought to be responsible for transduction, immunocytochemical studies were performed using polyclonal antibodies raised in rabbit against the alpha subunit of rat type I and type II voltage-gated sodium channels. The results show the presence of label on the neurite and axolemma, as well as in the node regions. Interestingly, labeling is also found on the inner and outer lamellae that form the non-neural accessory structure surrounding the neurite. The presence of this label in the surrounding lamellae suggests that voltage-sensitive Na+ channels, that are involved in both transduction and action-potential generation, may be made available to the neurite via transport from the lamellae, a mechanism perhaps operating in parallel to axoplasmic transport.

Pacinian corpuscle in the human pancreas.

During our systematic examination of the distribution of cytochrome P450 enzymes in the normal and diseased human pancreas, we observed a Pacinian corpuscle in a serial section of a tissue from a pancreatic cancer patient. We report the histologic and immunohistochemical patterns in this corpuscle and review the literature. The Pacinian corpuscle was situated within the pancreas of a 76-year-old woman with cancer in the head of the pancreas. We could demonstrate immunoreactivity within the corpuscle for the neurofilament protein. neuron-specific enolase, S-100 Protein, and for four cytochrome P450-isozymes. The possible function of Pacinian corpuscles in the mammalian and human pancreas is discussed.

Topography of corpuscular mechanoreceptors in the shoulder joint region of monodelphis domestica.

The topography and structure of corpuscular mechanoreceptors in the shoulder joint capsule and periarticular connective tissue of a small laboratory marsupial (monodelphis domestica) were studied using light and electron microscopy. This animal is known to use its upper extremities for a wide range of activities like climbing and manipulating food. Thus, the shoulder joint of this animal species has a similar wide range of movement as the human shoulder joint, but is small enough for serial sectioning in its entirety. Silver stained serial paraffin sections were examined under the light microscope and the distribution of the different types of mechanoreceptors was reconstructed using three-dimensional image processing. In addition, selected mechanoreceptors were studied electron microscopically. Approximately 100 small lamellated corpuscles were found in the dense connective tissue of the joint capsule close to the insertion on the scapula and in the thickening of the joint capsule close to the glenoid labrum. Ruffini corpuscles were found in much smaller numbers in the moderately dense connective tissue of the axillary region. Only very few Vater-Pacinian corpuscles were seen in the soft periarticular connective tissue. The large number and localization of mechanoreceptor corpuscles in the shoulder joint capsule especially close to the glenoid labrum suggests, that these specialized nerve endings are likely to play an important role in control of joint movement. They can induce protective reflexes during extreme movements in the shoulder joint preventing shoulder luxation by increasing the tone of muscles pressing the humerus head into the glenoid cavity.

A Pacinian corpuscle in the human bladder lamina propria.

The fortuitous finding of a complex Pacinian corpuscle within the lamina propria of the human urinary bladder is described. It consisted of a complex of encapsulated nerve endings within the areolar connective tissue of the lamina propria immediately adjacent to the inner aspect of the detrusor muscle. It showed no structural evidence of directional sensitivity and was associated on its outer aspect with small unmyelinated axons containing small clear and dense-cored vesicles. This appears to be the first report of an encapsulated nerve ending within the lining of the adult human urinary bladder.

Lectin and proteoglycan histochemistry of feline pacinian corpuscles.

We studied carbohydrate residues of glycoproteins and proteoglycans (PGs) in peritoneal Pacinian corpuscles of five adult cats. Terminal monosaccharides of glycoproteins and related polysaccharides were identified by lectin histochemistry and the PGs and glycosaminoglycans (GAGs) by specific antibodies. The most intensive lectin staining reactions indicated an abundance of glycoconjugates with terminal mannose (Man) or sialic acid residues, but no complex-type oligosaccharides were detected within the corpuscles. Terminal fucose (Fuc) and galactose (Gal) residues typical for O-linked mucin-type glycoproteins generally associated with high water binding capacity were also absent. Antibodies against unsulfated chondroitin (C-0-S), chondroitin-4-sulfate (C-4-S), and decorin showed positive reactions in the interfibrillar spaces between the lamellae, around collagen fibers, and around the lamellae of the perineural capsule, especially in the outer parts known to contain Type II collagen. Biglycan showed a preference for the innermost part of the perineural capsule (intermediate layer), known to contain Type V collagen. Collagen V and biglycan are both linked to growth processes. Hyaluronic acid (HA), chondroitin-6-sulfate (C-6-S) chains, and a chondroitin sulfate proteoglycan (CSPG) were co-localized in the terminal glia. The study of carbohydrates with high water binding capacity may contribute to our understanding of the high viscoelasticity of Pacinian corpuscles.

Restoration of lamellar structures in adult rat Pacinian corpuscles following their simultaneous freezing injury and denervation.

The capsule and inner core are multilamellar auxiliary structures enveloping the axon terminal of the Pacinian corpuscle. The freezing injury of the rat interosseal Pacinian corpuscles induced the destruction of all cellular components while the extracellular matrix including the basal laminae survive the treatment. Simultaneous denervation and the freezing treatment of the Pacinian corpuscles discovered an ability of the basal lamina and other components of the extracellular matrix to stimulate a differentiation of migrated Schwann cells and fibroblasts into multilamellar auxiliary structures. The restoration of inner core and capsule in the Pacinian corpuscles was independent of the presence of sensory axon terminals. The restored lamellar structures of Pacinian corpuscles in long-term surviving rat (4 to 8 months) displayed atrophic changes. The results suggest that the extracellular matrix of rat Pacinian corpuscles may contain molecules that are produced by Schwann cells and fibroblasts during maturation of the multilamellar auxiliary structures. The molecules deposited into the extracellular matrix are able to influence the redifferentiation of multilamellar auxiliary structures from immature cells.

Immunocytochemical identification of proteins within the Pacinian corpuscle.

Light- and electron-microscopic immunocytochemistry (ICC) was performed on Pacinian corpuscles (PCs) obtained from cat mesentery to determine the presence and location of various proteins within the accessory capsule and the neurite. Antibodies to tubulin, neurofilament 200, actin, collagen II and V, glial fibrillary acidic protein (GFAP) and S-100 were used. Type II collagen was localized only in the outer core of the accessory capsule, which is composed of an inner core, an intermediate layer or growth zone, an outer core and an external capsule. Type V collagen was found only in the intermediate growth zone. Intermediate filaments labeled with anti-GFAP were only found in the inner core. The calcium-binding protein that was labeled by anti-S-100 was found only in the inner core. Diffuse and variable staining for actin is present throughout the accessory capsule. The differences in distribution of these various proteins within the capsule suggest different structural/functional properties of the various capsule regions. The neurite was found to contain microtubules (i.e., tubulin) and neurofilaments throughout, but these cellular inclusions were not found within the cytoplasmic extensions (filopodia) that project from the neurite into the hemilamellar clefts formed by the inner-core hemilamellae. The extensions, however, were found to contain actin in a much greater density than that seen in the neurite proper. The presence of actin, but apparent lack of other cytostructural elements within the extensions, is highly reminiscent of the composition of stereocilia found on vestibular and auditory hair cells. Since stereocilia have been shown to play a role in hair-cell mechanotransduction, it is possible that the cytoplasmic extensions are significantly involved with mechanotransduction within the PC.

The extracellular matrix of rat pacinian corpuscles: an analysis of its fine structure.

The Pacinian corpuscle consists of a sensory axon terminal that is enveloped by two different structures, the inner core and the capsule. Since proteoglycans are extremely water soluble and are extracted by conventional methods for electron microscopy, the current picture of the structural composition of the extracellular matrix in the inner core and the capsule of the Pacinian corpuscle is incomplete. To study the structural composition of the extracellular matrix of the Pacinian corpuscles, cationic dyes (ruthenium red, alcian blue, acridine orange) and tannic acid were applied simultaneously with the aldehyde fixation. The interosseal Pacinian corpuscles of the rat were fixed either in 2% formaldehyde and 1.5% glutaraldehyde, with the addition of one of these cationic dyes or, in Zamboni's fixative, with tannic acid added. The cationic dyes and tannic acid revealed a different structural pattern of proteoglycans in the extracellular matrix in the inner core and in the capsule of the rat Pacinian corpuscles. The inner core surrounding the sensory axon terminal is a compartment containing proteoglycans that were distributed not only in the extracellular matrix but also in the cytoplasm of the lamellae. In addition, this excitable domain was separated from the capsular fluid by a thick layer of proteoglycans on its surface. An enlarged interlamellar space of the capsule contained large amounts of proteoglycans that were removed by digestion with chondroitinase-ABC. Ruthenium red and alcian blue provided only electron dense granules, probably corresponding to collapsed monomeric proteoglycan molecules. Acridine orange and tannic acid preserved proteoglycans very well and made it possible to visualize them as "bottlebrush" structures in the electron microscope. These results show that the inner core and the capsule of rat Pacinian corpuscles have different structural patterns of proteoglycans, which are probably involved in different functions.