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Ultrastructural localization of calcium in mechanoreceptors of the oral mucosa.

Cytochemical localization of Ca2+ in Meissner corpuscles and Merkel cell-neurite complexes in the palatine mucosa of the Mongolian gerbil was studied by a combined oxalate antimonate-microwave irradiation procedure. The reaction products obtained were identified as calcium antimonate by EGTA solubility and X-ray microanalysis. Meissner corpuscles in the normal palatine rugae could be roughly classified into three types by amount and localization of Ca2+. Type I corpuscles were characterized by a high Ca2+ content in both the terminal axoplasm and caveolae of the lamellar plates, type II, by a low Ca2+ content in the terminal axoplasm and a high Ca2+ content in the lamellar cytoplasm. Type III corpuscles showed intermediate characteristics. Palatine rugae stimulated mechanically during fixation contained an increased number of type I corpuscles. On the other hand, two patterns were distinguished in the distribution of Ca2+ in Merkel cells in palatine rugae fixed under normal conditions. One showed abundant Ca2+ dispersed throughout the cell, while in the other, Ca2+ was specifically localized in the Golgi apparatus and mitochondria. Similar distribution patterns also were observed in palatine rugae that had received mechanical stimulus during fixation. Axon terminals of most Merkel cell-neurite complexes in normal palatine rugae were poor in axoplasmic Ca2+, whereas those in most Merkel cell-neurite complexes in mechanically stimulated palatine rugae contained abundant Ca2+ in their axoplasm.

Animals↗

Mechanoreceptors of the hard palate of the Mongolian gerbil include special junctions between epithelia and Meissner lamellar cells: a comparison with other rodents.

As a part of our comparative morphological study of mechanoreceptors in rodent oral mucosae, the hard palate of the mongolian gerbil (a kind of sand rat) was investigated by light and electron microscopy. Subepithelial connective tissue papillae of palatine rugae contained a dense distribution of lamellated corpuscles that showed an ultrastructure similar to that of Meissner corpuscles of primates. The corpuscles in the palate of the mongolian gerbil, however, frequently formed compound large corpuscles with a palisade-like extension. Antemolar rugae predominated in containing such compound "Meissner" corpuscles. Superficial cytoplasmic lamellae of the corpuscles closely adhered to overlying epithelial cells without an intervening basal lamina. Although a small number of simple corpuscles was seen in the first antemolar rugae, no similar corpuscles were encountered in other parts of the palate. On the other hand, numerous Merkel cell-nerve endings were seen in epithelial pegs and sometimes in epithelial ridges of palatine rugae. Quinacrine labelling showed that intermolar rugae contained a rather dense distribution of Merkel cells in comparison with antemolar rugae. The present and previous data indicate that the pattern of palatine mechanoreceptors of the mongolian gerbil is analogous to that of the mouse, but not of the rat.

Animals↗

The intraepidermal innervation of the snout skin of the opossum. A light and electron microscope study, with observations on the nature of Merkel's Tastzellen.

The intraepidermal innervation of the snout skin of the opossum has been studied with the light and electron microscope. Numerous large nerve fibers loose their myelin sheath in the superficial dermis and pass into the epidermis. The basement membranes of the epidermis and Schwann cell become continuous at the point of entry of the neurite into the epidermis. Within the epidermis, the neurite is associated with a specialized secretory epidermal cell, termed a Merkel cell. This cell has many secretory granules apposed to the neurite. The Merkel cells are epidermal cells since they have desmosomes between them and adjacent epidermal cells. The neurite in the stratum spinosum is enveloped by Schwann cells in a manner analogous to the Schwann cell investment of unmyelinated neurites. In the upper stratum spinosum the nerve fiber evidences changes which can be interpreted as degenerative. The Merkel cell-neurite complex is interpreted as representing a sensory receptor unit.

Animals↗

Membrane properties of two types of basal cells in Necturus taste buds.

Necturus taste buds contain two types of basal cells: presumptive stem cells and Merkel-like basal cells. Both types of basal cells are small round cells located at the base of the taste bud, indistinguishable from each other with light microscopy. However, with electron microscopy, autoradiography, or immunocytochemistry, these two types of basal cells can be easily distinguished. We isolated basal cells from taste buds, characterized their voltage-dependent currents using gigaseal whole-cell recordings, and processed the cells for electron microscopy or immunocytochemistry. We were able to distinguish two cell types electrophysiologically and to correlate cell type with membrane properties. Isolated Merkel-like basal cells had several voltage-activated currents: transient, TTX-sensitive, inward Na+ current; sustained, saturating outward K+ current; and slowly inactivating inward Ca2+ current. These currents are similar to those observed in taste receptor cells. In contrast, presumptive stem cells from Necturus taste buds only had outward K+ currents.

Animals↗

The Haarscheibe.

The Haarscheibe is a specialized spot of epidermis containing many Merkel cell-neurite complexes. It is a highly sensitive, slowly adapting, modality-specific touch receptor occurring in all mammals. Its exact role in signaling sensation in human beings is undetermined, and the function of Merkel cells, with their distinctive cytoplasmic granules, in transducing mechanical force into neural action potentials remains to be determined.

Action Potentials↗

Carbonic anhydrase activity in axon terminals of sensory corpuscles.

The distribution of carbonic anhydrase (CA) activity was studied by electron microscopic histochemistry in rat Pacinian corpuscles, Meissner corpuscles and Merkel cell-neurite complexes using the cobalt bicarbonate method. The distribution of CA activity in these axon terminals was compared to the activity in sciatic nerve axons. An intense enzymatic CA activity was demonstrated in axon terminals of both Pacinian and Meissner corpuscles, while a weak activity was found within the axoplasm of terminals abutting Merkel cells. Some large- and medium-sized axons in sciatic nerves exhibited an intense activity. These findings indicate that large- or medium-diameter sensory axons innervating corpuscular endings have an intense CA activity extending from their somata to their sensory terminals. Axons to Merkel-neurite complexes differ in CA activity from those innervating Meissner and Pacinian corpuscular endings.

Animals↗

Special program of differentiation expressed in keratinocytes of human haarscheiben: an analysis of individual cytokeratin polypeptides.

Human haarscheiben, epidermal Merkel cell-rich sensory organs of hairy skin, were studied for the expression of various cytokeratin (CK) polypeptides and other epithelial and neuronal differentiation markers by applying immunoperoxidase and immunofluorescence microscopy to frozen sections and by two-dimensional gel electrophoresis. The basal clusters of Merkel cells were specifically detected by antibodies against CK 20. Haarscheiben keratinocytes were unique mainly by the prominent expression of CK 17 in the lower and middle layers. Further differences as compared to keratinocytes of usual epidermis included the enlargement of the basal compartment, characterized by the expression of CK 5 and the absence of the maturation-associated CKs 1/10/11, and the reduction of CK 15, which is a constituent of normal basal cells. Using CK 17 as a highly sensitive Haarscheibe marker in skin tissue sections, variabilities in the spatial relationship of the haarscheibe and the corresponding hair follicle were recorded. The results show that haarscheibe keratinocytes express a special program of differentiation that may be important for optimal stimulus perception. Immunohistochemical stainings for CK 17 will facilitate further studies on the distribution and biology of haarscheibe.

Adult↗

Primary trigeminal afferent neuron of the cat: II. Neuropeptide- and serotonin-like immunoreactivity.

The distribution of several peptides and serotonin in the cat vibrissa primary afferents from the Merkel cell-axon complexes to their central projections in the principal trigeminal sensory nucleus was examined by peroxidase-antiperoxidase immunocytochemical technique. In the trigeminal ganglia, two populations of primary sensory neurons, containing immunoreactive material were identified: a few substance P-, bombesin-, somatostatin-, cholecystokinin-, and vasoactive intestinal polypeptide-like immunoreactive ganglion cells with small and medium-sized somata, and many large non-labelled trigeminal neurons that were surrounded by substance P-, somatostatin-, cholecystokinin-, vasoactive intestinal polypeptide- and serotonin-positive pericellular baskets. Substance P-, bombesin- and vasoactive intestinal polypeptide-like immunoreactive substances were found in the basal side of Merkel cells from sinus hair follicles of the cat, whereas the associated axons were immunonegative. In the principal trigeminal sensory nucleus, cell bodies and fibres, containing substance P and bombesin were observed in both subdivisions. In addition, only nerve fibres and terminals showed somatostatin-, cholecystokinin-, vasoactive intestinal polypeptide- and serotonin-like immunoreactivity. The present results provide direct evidence that an ascending peptidergic system exist and that only substance P and, less likely, bombesin may be primary afferent neuroregulator(s) for the mechanosensory information in the cat trigeminal system.

Afferent Pathways↗

Sensory innervation of gingival and alveolar mucosa of the house musk shrew (Suncus murinus).

The lingual gingival and the alveolar mucosa of mandible of the house musk shrew (Suncus murinus) were stained by methylene blue vital staining or osmic acid staining, and mounted as whole thickness preparations. The sensory innervation and the distribution of sensory receptors were investigated with a light microscope. The nerve fibers supplying these regions derive from the sublingual nerve, which ascend in the mucosa as they branch out. Sensory receptors found in the present study are of four kinds; free nerve endings, bush-like nerve endings, Merkel cell-neurite complexes and encapsulated corpuscles. The Merkel cell-neurite complexes were scarce and localized in the upper margin of gingival mucosa. The bush-like nerve endings were distributed preferentially in the alveolar mucosa, in which their maximum density was 9-23 per mm2. Among the organized receptors, the encapsulated corpuscles appeared most frequently throughout the mucosal area investigated, and their maximum density amounted to 27-56 per mm2 in the gingival mucosa. These corpuscles were relatively small and poorly differentiated. Although the bush-like nerve endings and the encapsulated corpuscles were fewer in the third molar region, there was no obvious regional difference in their distribution densities from the premolar region to the second molar region.

Animals↗

Primary trigeminal afferent neuron of the cat: I. Studies on membrane-bound enzyme histochemistry.

The localization of some membrane-associated enzymes such as alkaline phosphatase, 5'-nucleotidase, glucose-6-phosphatase, Na+,K(+)-adenosine triphosphatase, adenylate cyclase and guanylate cyclase in the Merkel cell-axon complexes, trigeminal ganglia and the principal trigeminal sensory nucleus of the cat was determined at light and electron microscopic level using cytochemical techniques. In the sinus hair follicles (vibrissae), the reaction end product marking alkaline phosphatase and adenosine triphosphatase activities was visualized on the axons running through external follicle epithelium and the 5'-nucleotidase, adenylate- and guanylate cyclase positive reaction was seen to stain the plasma membranes of Merkel cells. In the trigeminal ganglia, the strongest alkaline phosphatase and adenosine triphosphatase activities showed the corresponding areas between the ganglion and satellite cells. 5'-nucleotidase activity was more intense on the neurilemmas and the surrounding glial plasma membranes. In the principle sensory trigeminal nucleus, the central neurons exhibited an intense alkaline phosphatase, 5'-nucleotidase and adenosine triphosphatase activities and much smaller amount of reaction product for adenylate cyclase and guanylate cyclase was observed. In conclusion, membrane-bound enzymes could be histo- and cytochemically demonstrated in all components of primary trigeminal afferent units. Our results have confirmed that the receptor function and the nerve impulses conductance need an intensive molecular and cation exchange, and energy supply.

5'-Nucleotidase↗

The innervation of the mystacial pad of the rat as revealed by PGP 9.5 immunofluorescence.

The innervation of the mystacial pad in the rat was investigated with the aid of antihuman protein gene product (PGP) 9.5 immunofluorescence. PGP 9.5 is ubiquitin carboxyl-terminal hydrolase, which is distributed throughout neuronal cytoplasm. This technique revealed all previously known innervation as well as a wide variety of small-caliber axons and some endings of large-caliber afferents that had not been observed before. Newly revealed innervation affiliated with vibrissal-follicle sinus complexes included 1) fine-caliber, radially oriented processes in the epidermal rete ridge collar; 2) a loose network of fine-caliber, circumferentially arrayed processes in the centrifugal part of the mesenchymal sheath at the level of the ring sinus; 3) a loose haphazard network of fine-caliber and medium-caliber processes in the mesenchymal sheath and among the trabeculae of the cavernous sinus; 4) a loose network of circumferentially arrayed processes within the mesenchymal sheath of the cavernous sinus and in close proximity to the basement membrane; 5) a dense network of reticular-like endings provided by large-caliber afferents to the mesenchymal sheath in the upper part of the cavernous sinus; and 6) fine-caliber innervation to the dermal papilla at the base of all vibrissal shafts. In the intervibrissal skin, a dense distribution of fine-caliber individual and clustered profiles was detected in the epidermis. In addition to previously known innervation, Merkel endings were consistently observed in the epidermis at the mouths of guard hairs, loose networks of fine-caliber axons were found around the necks of occasional guard hairs, and fine-caliber profiles were frequently affiliated with vellus hairs. Vascular profiles were heavily innervated throughout the dermis. Axons and motor end plates of the facial nerve innervation to papillary muscles also were labeled. Transection of the infraorbital nerve eliminated all but the facial nerve innervation. Unilateral removal of the superior cervical ganglion eliminated the innervation to the dermal papillae but caused no other noticeable reduction. PGP 9.5-like immunofluorescence was also moderately expressed in apparent Schwann cells, in Merkel cells only in the external root sheath of vibrissal follicles, and in apparent dendritic and/or Langerhans cells usually located in the epidermis and occasionally in the follicles. PGP 9.5-like immunofluorescence persisted in highly vacuolated profiles along the usual courses of medium to large-caliber axons 2 weeks after nerve transection. The possible functional role of the newly discovered innervation is considered along with that of previously identified afferents.

Animals↗

Regulation of NGF-family ligands and receptors in adulthood and senescence: correlation to degenerative and regenerative changes in cutaneous innervation.

During development, a highly differential neurotrophin dependency is reported for various types of nerve endings in the whisker follicle. To what extent these dependencies extend and play a role in adulthood is largely unresolved. We show here, using in situ hybridization and immunohistochemistry that the expression of neurotrophins and trk/p75 receptors persists in adulthood. As suggested by their expression profiles, many classes of cutaneous nerve endings disclose similar ligand-receptor dependencies in adult animals as during development, while other populations appear to switch their dependency. Furthermore, our data suggest that sensory endings that have a high turnover due to mechanical wear and tear, e. g. Merkel cell-neurite complexes at the level of ring sinus show a more complex ligand-receptor expression phenotype than do endings with a less vulnerable location, e.g. the Merkel cell-neurite complexes at the rete ridge collar. Thus, neurotrophin-3 (NT3)/trkA signalling is suggested to be important for a continuous terminal plasticity of Merkel cell-neurite complexes at the level of ring sinus in adulthood. Evidence supporting a role for neurotrophin signalling in maintaining the adult cutaneous innervation also comes from the close correlation between altered ligand-receptor expression(s) and axonal/terminal aberrations in senescence. Thus, an ageing-related decrease in target neurotrophin expression, notably NT3 and NT4, results in a site-specific loss of sensory terminals concomitant with an aberrant growth of regenerating/sprouting axons into new target fields. Ageing of the cutaneous innervation, manifested in degenerative and regenerative events, seems strongly associated with changes in neurotrophic interactions between sensory neurons and target tissues.

Aging↗

Villin is a possible marker of receptor cells in frog taste organs.

We investigated lingual taste organs of four frog species mainly by means of fluorescence immunohistochemistry for villin, calbindin, and serotonin. Cells immunoreactive for villin appeared in the taste organs of all the species used. These villin-immunostained cells were basoapically elongated in shape and extended up to the apical surface. They were also immunoreactive for calbindin. On the other hand, serotonin-immunoreactive cells, identified as Merkel-like basal cells, were immunonegative for villin. Considering the present results combined with those of studies by other research groups, the villin-immunostained cells were postulated to function as taste receptors.

Animals↗

Beta-mannosidosis: lesions of the distal peripheral nervous system.

Beta-Mannosidosis, an inherited glycoprotein metabolic disorder so far identified only in the caprine species, is characterized by deficiency of beta-mannosidase, oligosaccharide accumulation and excretion, neurovisceral cytoplasmic lysosomal storage vacuoles and central nervous system myelin and axonal lesions. In this investigation, transmission electron microscopy was used to study peripheral nerve elements in the gingiva of five affected goats and a control animal. Merkel and Schwann cells were vacuolated. Accumulations of dense bodies distended Merkel cell end plates and free-ending axons in the prickle cell layer as well as Pacinian corpuscle axons, and myelinated, unmyelinated, and demyelinated axons in the lamina propria. Unlike central nervous system myelin paucity, loss of peripheral nerve myelin was exclusively related to axonal dense body accumulation and enlargement. The identification of these lesions may facilitate detection of beta-mannosidosis in man and other species.

Animals↗

Effects of trifluoperazine and amiloride on the extrusion of axoplasmic Ca++ by rapidly adapting and slowly adapting type I cutaneous mechanoreceptors: an electron-microscopic study using the oxalate-antimonate method.

Influences of Ca++ transport inhibitors on the extrusion of cytosolic Ca++ from axon terminals of oral mucosal mechanoreceptors were cytochemically studied through the use of trifluoperazine (TFP), a Ca++ pump inhibitor, and amiloride, a Na+/Ca++ exchanger inhibitor. Palates of Mongolian gerbils were isolated after perfusion with normal, TFP-containing, or amiloride-containing HEPES-buffered saline, and then mechanically stimulated in the same buffer. Cytosolic Ca++ in axon terminals at 2 min after the stimulation was visualized by means of an oxalate-antimonate method using microwave fixation, and then evaluated electron-microscopically. TFP was found to inhibit the Ca++ extrusion from axon terminals of Meissner corpuscles, but not from those of Merkel cell-axon complexes. Conversely, amiloride severely reduced the Ca++ extrusion from axon terminals of Merkel cell-axon complexes, but not from those of Meissner corpuscles. These results suggest that cytosolic Ca++ extrusion from axon terminals of rapidly adapting and slowly adapting type I mechanoreceptors is regulated by a Ca++ pump and an Na+/Ca++ exchanger, respectively.

Amiloride↗