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PubMed2003

Incidence of cytokeratin 20 expressed cells in primitive follicular structure and secondary neoplastic proliferations of nevus sebaceus.

BACKGROUND: Nevus sebaceus of Jadassohn (NSJ) is a complex hamartoma in which various kinds of secondary neoplastic or hyperplastic proliferations such as primitive follicular structure (PFS) may arise. Recently, cytokeratin (CK) 20 expressed cells in PFSs and trichoblastomas of NSJs were demonstrated. OBJECTIVE: To clarify the nature and relationship among these proliferations, those in other secondary neoplasms and precise CK distributions in such lesional epidermis are examined. METHODS: 49 cases of NSJ were analyzed clinicopathologically and immunohistochemically using ten monoclonal antibodies against involucrin and CKs. RESULTS: 15 of 49 cases possessed PFSs, and six cases, of which five were associated with PFSs, possessed seven lesions of secondary neoplasms: four BCC, two syringocystadenoma papilliferum (SAP) and one trichilemmoma. The age distribution of 15 patients with PFSs was significantly higher than that of the other 34 patients without the PFSs. The six patients with secondary neoplasms were statistically situated in a higher age range than the ten cases with the PFSs alone. Although the CK expression in lesional epidermis of NSJs were similar to that of normal epidermis, CK20 expressed cells were scattered in the PFSs of 11 cases among the 15 NSJs, but not in the other four cases. Three of the four cases were associated with BCC in which CK20 was not expressed. CONCLUSIONS: From the statistical analysis, we presumed that the existence of the PFSs indicates potency to develop secondary neoplasms. Moreover, the absence of CK20 expressed cells (Merkel cells) in the structures was speculated to predict the development of BCC in the future.

Adolescent↗
PubMed2004

Human hair greying is linked to a specific depletion of hair follicle melanocytes affecting both the bulb and the outer root sheath.

BACKGROUND: Although hair greying is a very common phenomenon characterized by loss of pigment in the hair shaft, the events that cause and control natural hair whitening with age in humans are still unclear. OBJECTIVES: To decipher the origin of natural hair whitening. METHODS: Human hair melanocytes were immunohistochemically characterized at different stages of whitening. RESULTS: Loss of hair shaft melanin was found to be associated with a decrease in both bulb melanin content and bulb melanocyte population. Although few melanocytes were present in the bulbs of grey hair, they still expressed tyrosinase and tyrosinase-related protein-1, synthesized and transferred melanins to cortical keratinocytes as seen by the presence of melanin granules. In white hair bulbs, no melanocytes could be detected either with pMel-17 or vimentin labelling. Pigmented hair follicles are known to contain inactive melanocytes in the outer root sheath (ORS), and grey and white hairs were also found to contain some of these quiescent melanocytes. However, their population was decreased compared with pigmented hair follicles, ranging from small to nil. This depletion of melanocytes in the different areas of white hairs was detected throughout the hair cycle, namely at telogen and early anagen stages. In contrast, the infundibulum and sebaceous gland of both pigmented and white hairs showed a similar distribution of melanocytes. Furthermore, other distinct cell populations located in the ORS, namely putative stem cells, Merkel cells and Langerhans cells were equivalently identified in pigmented and white hairs. CONCLUSIONS: Thus, hair greying appears to be a consequence of an overall and specific depletion of bulb and ORS melanocytes of human hair.

Adult↗
PubMed1985

Morphological and phenotypical characterization of bone marrow-derived dendritic Thy-1-positive epidermal cells of the mouse.

Increasing evidence exists that the spectrum of dendritic cells within the epidermis is more complex than previously thought. In addition to Langerhans cells, Merkel cells, and melanocytes, the murine epidermis contains a dendritic cell population whose most prominent phenotypic feature is the Thy-1 antigen. These cells are now generally referred to as dendritic Thy-1+ epidermal cells (dThy-1+EC). The ultrastructural features of these cells do not resemble those of other resident epidermal cells (EC). In particular, their cytoplasm contains abundant intermediate-sized filaments of the vimentin type as well as membrane-limited organelles with a central granular core. The bone marrow derivation of dThy-1+EC is now well established: dThy-1+EC carry Ly-5 determinants whose expression is restricted to cells of the hemopoietic differentiation pathway, and studies using Thy-1-disparate radiation bone marrow chimeras have revealed the presence of donor-type Thy-1+ cells within the epidermis; by immunoelectron microscopy, these cells represent dThy-1+EC. dThy-1+EC repopulate the epidermis at a slower rate than Langerhans cells as evidenced by a direct comparison of the repopulation kinetics of both cell systems in radiation bone marrow chimeras, and by experiments studying the emergence of either Ia+- or dThy-1+EC in an epidermis which had been previously depleted of either Langerhans cells (glucocorticosteroids) or of dThy-1+EC (PUVA). The phenotypical features of dThy-1+EC differ from those of thymus-derived lymphocytes, B cells, dendritic cells, and mononuclear phagocytes. The surface marker repertoire of dThy-1+EC (Thy-1, Ly-5, asialo-GM1) resembles certain members of the rather heterogeneous natural killer (NK) cell system but functional studies are needed to ascertain this contention.

Animals↗
PubMed1982

Separation of human skin cells by velocity sedimentation into functionally distinct fractions.

The epidermis consists of a heterogeneous population of cells including Langerhans cells, Merkel cells, melanocytes, and keratinocytes in various stages of differentiation. The current study was undertaken to determine if skin cell suspensions can be separated into morphologically and/or functionally distinct fractions. Skin cells were suspended by trypsinization and separated into multiple fractions by velocity sedimentation. Certain fractions reproducibly stimulated proliferation of allogeneic lymphocytes in the skin cell lymphocyte reaction, whereas other fractions, containing larger cells, supported growth of keratinocyte colonies in cell cultures. These results indicate that stimulation in the skin cell lymphocyte reaction and growth of keratinocyte colonies are mediated by distinct cells, separable by velocity sedimentation.

Cell Division↗
PubMed1969

The structure and function of a slowly adapting touch corpuscle in hairy skin.

1. Slowly adapting cutaneous mechanoreceptors, in the cat and primates, have been studied by histological and neurophysiological methods.2. Each touch corpuscle is a dome-shaped elevation of the epidermis, whose deepest layer contains up to fifty specialized tactile cells.3. Nerve plates, enclosed by the tactile cell (Merkel cells), are connected to a single myelinated axon in the dense collagenous core of the corpuscle.4. The corpuscle generated > 1000 impulses/sec when excited by vertical surface pressure. The response was highly localized and showed a low mechanical threshold, the frequency being dependent upon the velocity and amplitude of the displacement. There was a period of rapid adaptation before a sustained response which might continue for > 30 min.5. A quantitative analysis of the responses to excitation by displacements of differing amplitude, velocity and duration is included.6. The discharge of touch corpuscle units evoked by a mechanical stimulus was temperature-sensitive, and was enhanced by a fall in skin temperature.

Adaptation, Physiological↗
PubMed2000

Morphological changes in the skin during experimental allergic encephalomyelitis and multiple sclerosis: bases for new diagnostic test.

Skin specimen from the medial malleolar area of patients with multiple sclerosis and guinea pigs with experimental allergic encephalomyelitis were examined by light and electron microscopy. Demyelination, inflammation, and dystrophic changes in peripheral nerves and skin correlating with clinical manifestations were revealed in both sclerosis and encephalomyelitis. Maximum changes were revealed in nerve-associated cells (Merkel cells), Langerhans cells reflecting inflammation intensity, and fibroblasts responsible for nerve integrity and resistance against external stimuli.

Adult↗
PubMed2003

Lighting up the senses: FM1-43 loading of sensory cells through nonselective ion channels.

We describe a novel mechanism for vital fluorescent dye entry into sensory cells and neurons: permeation through ion channels. In addition to the slow conventional uptake of styryl dyes by endocytosis, small styryl dyes such as FM1-43 rapidly and specifically label hair cells in the inner ear by entering through open mechanotransduction channels. This labeling can be blocked by pharmacological or mechanical closing of the channels. This phenomenon is not limited to hair cell transduction channels, because human embryonic kidney 293T cells expressing the vanilloid receptor (TRPV1) or a purinergic receptor (P2X2) rapidly take up FM1-43 when those receptor channels are opened and not when they are pharmacologically blocked. This channel permeation mechanism can also be used to label many sensory cell types in vivo. A single subcutaneous injection of FM1-43 (3 mg/kg body weight) in mice brightly labels hair cells, Merkel cells, muscle spindles, taste buds, enteric neurons, and primary sensory neurons within the cranial and dorsal root ganglia, persisting for several weeks. The pattern of labeling is specific; nonsensory cells and neurons remain unlabeled. The labeling of the sensory neurons requires dye entry through the sensory terminal, consistent with permeation through the sensory channels. This suggests that organic cationic dyes are able to pass through a number of different sensory channels. The bright and specific labeling with styryl dyes provides a novel way to study sensory cells and neurons in vivo and in vitro, and it offers new opportunities for visually assaying sensory channel function.

Animals↗
PubMed2005

Expression of human beta-defensin-3 in gingival epithelia.

OBJECTIVE: This study aimed to investigate the expression patterns of the newly discovered human beta-defensin-3 (hBD-3) in human gingiva. BACKGROUND: Human beta-defensins (hBDs) are a group of small, broad-spectrum, cationic antimicrobial peptides. Our recent study showed that the expression levels of hBD-1 and 2 peptides were associated with periodontal conditions. METHODS: A total of 49 gingival biopsies were collected, including 33 samples from 21 patients with chronic periodontitis and 16 samples from 16 periodontally healthy subjects. The expression of hBD-3 was detected by immunohistochemistry and in situ hybridization. Double staining was undertaken to identify hBD-3 peptide-positive cells, using CD-1a and cytokeratin 20 as markers for Langerhans cells and Merkel cells, respectively. RESULTS: hBD-3 peptide was detected in 88% of the samples, which was confined to the gingival epithelia. In healthy control subjects, hBD-3 peptide was more frequently detected in the basal layer as compared to the patients (53% vs. 18%, p < 0.05). In patients, hBD-3 expression extended from the basal layer to the spinous layers (82%), in which hBD-3 was confined to the basal and deep spinous layers in clinically healthy tissues from patients, whereas it extended to the superficial spinous layers in pocket tissues from patients (0% vs. 50%, p < 0.05). In both groups, hBD-3 peptide was expressed not only in gingival keratinocytes, but also in Langerhans cells and Merkel cells. hBD-3 transcripts were detected in 90% of the samples and they were confined to the basal and/or suprabasal layers of gingival epithelia. CONCLUSIONS: This study shows that hBD-3 is frequently expressed in gingival epithelia. The appropriate expression of hBD-3 peptide may contribute to the maintenance of periodontal homeostasis, possibly through its antimicrobial effect and promotion of adaptive immune responses.

Adolescent↗
PubMed2006

Merkel cell-nerve cell interaction undergoes formation of a synapse-like structure in a primary culture.

Merkel cells have been assumed to guide nerve fibers to the skin. However, there has been little in vitro evidence that supports this hypothesis, because there is no suitable established culture system of Merkel cells. Here we show that Merkel cells isolated from rat footpad skin were successfully cultured in a monolayer with keratinocytes. Keratinocytes did not affect any structural changes in Merkel cells. When nerve cells (NG108-15 or PC12) were added to the culture system, both nerve fibers and cytoplasmic processes of Merkel cells outgrew and cooperatively organized synapse-like structures at their contact points. Nerve cells promoted Merkel cell survival, compared with keratinocytes only. Merkel cell proliferation was not detected in all conditions, even with nerve growth factor, neurotrophin-3, interleukin-6 and tumor necrosis factor-alpha. The data suggest, firstly, that Merkel cells may guide nerve fibers to the skin by interacting with nerve cells; and, secondly, that nerve cells, but not keratinocytes, may produce some survival factors other than the cytokines above for Merkel cells, although Merkel cells may be a terminally differentiated cell type. Our method could open a way to study Merkel cell biology.

Animals↗
PubMed2004

The adult hair follicle: cradle for pluripotent neural crest stem cells.

This review focuses on the recent identification of two novel neural crest-derived cells in the adult mammalian hair follicle, pluripotent stem cells, and Merkel cells. Wnt1-cre/R26R compound transgenic mice, which in the periphery express beta-galactosidase in a neural crest-specific manner, were used to trace neural crest cells. Neural crest cells invade the facial epidermis as early as embryonic day 9.5. Neural crest-derived cells are present along the entire extent of the whisker follicle. This includes the bulge area, an epidermal niche for keratinocyte stem cells, as well as the matrix at the base of the hair follicle. We have determined by in vitro clonal analysis that the bulge area of the adult whisker follicle contains pluripotent neural crest stem cells. In culture, beta-galactosidase-positive cells emigrate from bulge explants, identifying them as neural crest-derived cells. When these cells are resuspended and grown in clonal culture, they give rise to colonies that contain multiple differentiated cell types, including neurons, Schwann cells, smooth muscle cells, pigment cells, chondrocytes, and possibly other types of cells. This result provides evidence for the pluripotentiality of the clone-forming cell. Serial cloning showed that bulge-derived neural crest cells undergo self-renewal, which identifies them as stem cells. Pluripotent neural crest cells are also localized in the back skin hair of adult mice. The bulge area of the whisker follicle is surrounded by numerous Merkel cells, which together with innervating nerve endings form slowly adapting mechanoreceptors that transduce steady skin indentation. Merkel cells express beta-galactosidase in double transgenic mice, which confirms their neural crest origin. Taken together, our data indicate that the epidermis of the adult hair follicle contains pluripotent neural crest stem cells, termed epidermal neural crest stem cells (eNCSCs), and one newly identified neural crest derivative, the Merkel cell. The intrinsic high degree of plasticity of eNCSCs and the fact that they are easily accessible in the skin make them attractive candidates for diverse autologous cell therapy strategies.

Animals↗
PubMed1977

The ultrastructure of cutaneous type I mechanoreceptors (Haarscheiben) in cats following denervation.

Denervation of specialized cutaneous mechanoreceptors (Haarscheiben or domes) in cats was followed after 20 and 25 days by the following alterations in receptor structure: (1) reduced numbers of Merkel cells, (2) Merkel cells degenerating in situ, (3) fewer dense-core granules in the cytoplasm of Merkel cells, (4) an increased number of agranular dendritic cells and Langerhans cells in the dome, (5) the apparent phagocytosis of Merkel cells by Schwann and Langerhans cells, (6) fewer epithelial cell layers over the dome, and (7) a decrease in the number of transitional cells. Skin excised between the domes in the denervated nerve field appeared normal when compared to innervated skin, and it was considered unlikely that the alterations in dome structure were due to generalized nutritional changes in the skin caused by transection of sympathetic axons or to some other side effect of denervation. Since domes are formed in new locations on the skin after nerves have regenerated (Burgess et al., '74), changes in dome structure following nerve transection are probably due to loss of the "trophic" influence of the nerves supplying the dome.

Animals↗
PubMed2001

The spinal nerves innervate putative chemosensory cells in the ventral skin of desert toads, Bufo alvarius.

Toads normally obtain water by absorption across their skin from osmotically dilute sources. When hyperosmotic salt solutions are presented as a hydration source to dehydrated desert toads, they place the ventral skin onto the source but soon afterwards escape to avoid dehydration. The escape behavior coincides with neural excitation of the spinal nerves that innervate putative chemosensory cells in the ventral skin. In the present study, fluorescent dye translocated through the spinal nerves to those receptor cells in the epidermis was photoconverted in the presence of 3, 3'-diaminobenzidine tetrahydrochloride for electron-microscopic observation of the cells and associated nerve terminals. Most of the photoconverted cells were located in the deepest layer of the epidermis, with some being in more intermediate layers. No labeled cell was seen in the outermost layer of living cells. In desert toads, flask cells and Merkel cells are occasionally seen in the epidermis. An association of nerve fibers with these epidermal cells has been reported in some species of the anurans. In the present study, however, the cytological features of the photoconverted cells are neither reminiscent of flask cells nor Merkel cells, but are similar to those of surrounding epithelial cells in each layer of the epidermis. We hypothesize a sensory function for these cells, because they have a close association with nerve fibers and participate in the transepithelial transport of salts that must pass through all cell layers of the skin.

Animals↗
PubMed2000

An immunohistological study of cytokeratin 20 in human and mammalian oral epithelium.

Cytokeratin (CK) 20 is a low molecular-weight intermediate filament reportedly expressed only by benign and malignant gastrointestinal epithelium, urothelium and Merkel cells. The main aims here were to map its expression in normal oral mucosa of humans and other mammals, and to determine whether it was expressed by abnormal human oral epithelium. Salivary and odontogenic epithelium were also analysed. An immunoperoxidase method was used on wax-embedded and cryostat sections. In addition, double-labelling experiments were undertaken to determine the association between CK 20 expression and that of CK 8/18 or S100 protein. Normal human oral mucosa from four sites, together with abdominal skin, was studied in autopsy samples from 32 individuals. CK 20-positive, basally situated, round or angular cells, consistent with Merkel cells, were recorded in 24/32 (75.0%) samples of mandibular gingiva, 25/32 (78.1%) samples of hard palate, 7/32 (21.9%) samples of buccal mucosa, 0/32 samples of lateral border of tongue, and 2/32 (6.3%) samples of abdominal skin. Double-labelling showed that all CK 20-positive Merkel cells also expressed CK 8/18 and S100. The only other cells to express CK 20 were human taste buds. There was no expression by dysplastic or invasive oral epithelium from biopsy samples. Colonic mucosa showed luminal-cell positivity in man, marmoset, ferret, rabbit and guinea-pig, but oral mucosa was universally negative in non-human species. It is concluded that in oral mucosa CK 20 is a specific marker of Merkel cells and taste buds, that Merkel cells are more frequently present in keratinized than non-keratinized oral mucosa, that CK 20-positive Merkel cells are also S100-positive, that there may be interspecies variations in CK 20 polypeptide composition and that, by contrast to urothelium, CK 20 has no value in the diagnosis of oral epithelial dysplasia.

Adult↗
PubMed1988

Immunological studies on the occurrence and properties of chromogranin A and B and secretogranin II in endocrine tumors.

We investigated a variety of endocrine tumors for the presence of chromogranins A and B and secretogranin II. These antigens were identified by one- and two-dimensional immunoblotting and in some cases by immunohistochemistry. An antigen corresponding in electrophoretic behavior to adrenal chromogranin A was present in all types of tumors, including insulinomas, oat cell carcinomas, and Merkel cell tumors of the skin. Chromogranin B had a much more limited distribution. This antigen could not be detected in parathyroid adenomas, oat cell carcinomas, or Merkel cell tumors, either by immunoblotting and immunohistochemistry. The occurrence of secretogranin II was similar to that of chromogranin B, with the exception of a positive reaction in Merkel cell tumors. In benign pheochromocytomas, all three antigens were found consistently; whereas in two of three malignant pheochromocytomas, chromogranin B was absent. Our study establishes that in most cases chromogranins and secretogranin in tumors are identical to the adrenal antigens, but that these antigens are not always stored together. Chromogranin A is the most widely distributed marker for endocrine tumors.

Adrenal Gland Neoplasms↗
PubMed1994

Merkel's cell (neuroendocrine) carcinoma of the vulva.

BACKGROUND: Merkel's cell carcinoma rarely arises in the vulva. It is unclear whether those cases arising in the vulva behave differently from the usual Merkel's cell carcinomas. METHODS: A Merkel's cell carcinoma of the vulva was studied by light microscopy, immunohistochemistry, and electron microscopy. The clinical data of this case and of other reported cases are summarized and compared with those of Merkel's cell carcinoma in general. RESULTS: This tumor showed the characteristic trabecular pattern of Merkel's cell carcinoma. The tumor cells were immunoreactive to low-molecular-weight cytokeratin and neuron-specific enolase. This patient was treated initially with local excision. She died 17 months later with progressive metastatic disease unresponsive to chemotherapy. This case study and the review of six other cases indicate that Merkel's cell carcinoma of the vulva is universally metastatic, both to the regional lymph nodes and distant sites, and that it invariably follows a rapidly fatal course. CONCLUSION: Merkel's cell carcinoma of the vulva appears to have a biologic behavior more aggressive than that of Merkel's cell carcinoma in general. An initial modality combining chemotherapy, with or without irradiation, with aggressive surgery should be tried in future cases.

Adult↗
PubMed2001

Immunohistochemical expression of G protein alpha-subunit isoforms in rat and monkey Merkel cell-neurite complexes.

The true function of Merkel cells (MCs) is still enigmatic, though the localization of various kinds of neurotransmitter-like substances in MCs has been revealed by immunohistochemistry. Most of the neurotransmitters act on target cells via seven-transmembrane receptors coupled to heterotrimeric G proteins. The heterotrimeric G proteins include various subfamilies that contribute to different signal transduction pathways. Therefore investigation of specific types of G proteins in MCs and related axon terminals (MC-axon terminals) should contribute to the elucidation of the function of MCs. In this study, we investigated the expression patterns of alpha-subunit isoforms of G proteins in MC-neurite complexes of the rat and monkey by enzymatic and fluorescence immunohistochemistry. MC-axon terminals of the rat and monkey showed positive immunoreactions of Galphao and Galphai1. Those of the monkey also showed a weak immunoreaction of Galphas. On the other hand, MCs of both animals showed positive immunoreactions of Galphao, Galphai1, Galphaq, and Galphaz. In addition, MCs of the monkey showed weak immunoreactions of Galphas. Galphao- and Galphai1-like immunoreactions in the MC-axon terminals suggest that MCs suppressively regulate receptive functions of type I mechanosensory nerve terminals. On the other hand, the localization of Galpha-subunits in MCs suggests that these cells are regulated with hormones, neurotransmitter-like substances, or growth factors.

Animals↗
PubMed1993

Epithelial skirt and bulge of human facial vellus hair follicles and associated Merkel cell-nerve complex.

Many morphological variations of bulge areas, such as knob-like swellings, were found in extracted human facial vellus hairs. In anagen vellus hair of the face bulge areas including these knobs had a band-like dense aggregation of CAM5.2 (K8, 52.5 kDa) reactive Merkel cells. In telogen hair the bulge became indistinguishable from the clubbed or regressed end of the follicle but Merkel cells continued to be abundant. The epithelial hood at sebaceous gland level showed most commonly a skirt-like structure but variations were also observed; these were bamboo joints, tulip flower, and long apron configurations. Merkel cells were found sparsely in these structures. Palisading stockade-like nerve endings were observed surrounding the follicular epithelium under the skirt and around the bulge areas including the knobs. Merkel cells were sparse in the follicular segment corresponding to the attachment of stockade nerve endings.

Adult↗
PubMed1980

Fine structural analysis of the synaptic junction of Merkel cell-axon-complexes.

The ultrastructure of synaptic contact areas in Merkel cell-axon-complexes from sinus hair follicles and touch domes of various mammals (nude mice, rats, cats, rabbits, opossums and monkeys) was investigated by electron microscopy of ultrathin sections from perfusion fixed tissue. Synapses between Merkel cells and axons were a common feature in all analyzed species. Special staining with digallic acid and goniometric tilting facilitated the resolution of the membranous and paramembranous synaptic elements. The synaptic contact revealed the typical characteristics of a chemical synapse, except for presynaptic clear vesicles: a postsynaptic membrane thickening and dense projections at the presynaptic membrane (i.e., the Merkel cell membrane). The cleft material was resolved as a fuzzy coating of the outer leaflets of the synaptic membranes with occasional bridges across the synaptic cleft. The presence of a synapse in the Merkel cell-axon-complexes emphasizes the receptor function of the Merkel cell besides other possible functions of this cell.

Animals↗