Size of desmosomes and hemidesmosomes in normal human epidermis

The dimensions of desmosomes and hemidesmosomes in normal human epidermis, i.e. their length and thickness, were studied in ordinary ultrathin sections. The length of desmosomes in the inner and middle squamous cell layer was 310 nm and 300 nm, while the thickness was 31 nm and 30 nm, respectively. In the outer squamous cell layer the values increased to 340 nm and 35 nm, respectively. The percentage total desmosomal length in relation to the perimeter of a cell showed the same tendency.     

Ultrastructural localization of cell junctional components (desmoglein, plakoglobin, E-cadherin, and β-catenin) in Hailey-Hailey disease, Darier's disease, and pemphigus vulgaris

The distribution of desmoglein, plakoglobin, E-cadherin, and β-catenin in the peri-lesional and lesional skin of Hailey-Hailey disease, Darier's disease, and pemphigus vulgaris was examined by immunoelectron microscopy. In the peri-lesional skin, the immunolabeling of these desmosomal components was localized to desmosomes. Adherens junction-associated E-cadherin and β-catenin were at the cell periphery, excluding desmosomes. The labeling pattern was similar among these diseases, but the labeling intensity particularly that of plakoglobin in Hailey-Hailey disease and Darier's disease, was less than that of normal controls, suggesting that these glycoproteins are quantitatively less concentrated in the normal epidermis of these inherited diseases. In the acantholytic cells of Hailey-Hailey disease and Darier's disease the immunolabeling of the components of desmosomes was diffusely distributed in the cytoplasms, whereas that of adherens junction was mostly at the cell periphery and partly diffusely in the cytoplasm. In contrast, desmosomes of detaching keratinocytes in pemphigus vulgaris still showed the labeling of desmoglein and plakoglobin. These findings suggest that the inherited acantholytic diseases, i.e., Hailey-Hailey disease and Darier's disease have a different pathogenesis from that of autoimmune acantholysis in pemphigus vulgaris: The intracellular components of desmosomes may primarily be disrupted in the genetic acantholytic diseases in the initial stages of acantholysis. Several unsolved questions in the previous light microscopic immunofluorescence sttidies using the same antibodies are now answered: 1) the diffusion of desmosomal proteins is not due to the internalization of desmosomes, 2) intracellular components of adherens junction are also finally dissolved, 3) diffuse cytoplasmic immunofluorescence patterns of desmosomal components could be explained by immunoelectron microscopy as those attached to cell membrane and trapped in tonofilament aggregates.     

Langerhans' Cells of Elderly Patients with Decubital Ulcers

An ultrastructural and morphometric study compared Langerhans' cells in sacral epidermis, 8–10 cm from the lesion of patients (mean age 70) with decubital ulcers, with those in the patients' own normal epidermis from the upper leg and with those in the epidermis from the upper leg of normal age-matched volunteers. The Langerhans' cell section area was significantly lower in patients' control leg epidermis (25.86 ± 2.29 μm²) than in that from normal controls (35.74 ± 3.76 μm²) or that from patients' sacral epidermis near the lesion (41.26 ± 3.45). The number of Langerhans' cell granules was higher in control leg epidermis of patients (10.22 ± 1.26) and was significantly higher in lesioned sacral epidermis (12.94 ± 1.90) than in normal controls (6.97 ± 1.47). In Langerhans' cells in patients' epidermis, formative stages of Langerhans' cell granules were observed. The findings may indicate that Langerhans' cells in patients' epidermis are in an active state.     

X-RAY EMISSION MICROANALYSIS OF THE DENSITY OF HAIR PROTEINS IN KWASHIORKOR

SUMMARY. Hair samples were obtained from the scalps of 5 infants with kwashiorkor and 5 matched control infants on a balanced diet.
The greatest diameters of the hairs were measured in their natural state, ethanol, xylol, water and air. The diameters of the kwashiorkor hairs were 8 to 22% larger in water than in air while the diameters of the control hairs were 5 to 8% larger. This indicates that the stability of the cross links between the polypeptide chains of hair proteins is reduced in kwashiorkor.
The mass per unit area of cryostat sections of hair was estimated by electron probe x-ray emission microanalysis. No difference was demonstrable between the density of proteins in kwashiorkor and control hair. It is concluded that the soft texture of kwashiorkor hair is not associated with a reduction in the density of its proteins.     

Sugars protect desmosome and corneosome glycoproteins from proteolysis

Summary Adhesional glycoproteins of desmosomes possess asparagine-linked, complex oligosaccharide side chains. We investigated the potential of these sugars to protect the core proteins of desmosomes and corneosomes (modified stratum corneum desmosomes) against proteolysis. Isolated pig ear epidermis was exposed sequentially to individual hydrolases, and their effect monitored ultrastructurally. Two major steps were employed: (1) glycosidases, to remove stepwise the sugars in a typical complex oligosaccharide chain; and (2) proteolysis using both endopeptidases and an exopeptidase. Controls were exposed to the same sequence of buffers, but without enzymes. Proteases alone induced no major changes in desmosomes or corneosomes compared with controls. Glycosidases alone, or proteases followed by glycosidases, caused mild fragmentation of the desmosomal interspace, but no widening. However, dramatic changes occurred when glycosidase treatment was followed by proteolysis. The interspace of both desmosomes and corneosomes was extensively digested, and consequently widened, causing loose packing of the epidermis. These findings indicate that sugars are potentially anti-proteolytic in both desmosomes and corneosomes. Sugars may, therefore, be a factor in preventing premature desquamation, by protecting desmosomes and corneosomes against extracellular proteases derived from membrane-coating granules.     

ELECTRON MICROSCOPIC STUDY OF THE EFFECT OF VITAMIN A ON THE DIFFERENTIATION OF RECONSTRUCTED EMBRYONIC CHICK SKIN

In reconstructed embryonic chick skin, the mass of epidermal cells was separated from the mesenchymal portion by a continuous basallamina. The findings on the cytoplasmic organelles, e.g. filamentous structures, desmosomes, keratohyalin, and membrane coating granules of the peripheral and central epidermal cells, corresponded to those of the epidermis when the centre of the cell-mass was presumed to he the surface of the epidermis. The addition of 5 i.u./ml. vitamin A in the culture medium inhibited the process of keratinization. Filamentous structures and desmosomes were sparse even in the centre of the epidermal cell-mass. There were wide intercellular spaces and the basal lamina was obscure and discontinuous. The endoplasmic reticulum was dilated and the Golgi zones were distinct. Thus the vitamin A-treated cell resembles the secretory cell and shows less tendency to differentiate towards a keratinocyte. The development of lysosomes in the vitamin A-treated cells confirmed that vitamin A releases protease.     

Morphometric characterization of the human neuroendocrine Merkel cells

In this study, the neuroendocrine Merkel cells (NEMCs) from adult human epidermis are defined morphometrically, using the MOP 20 image analyzer to measure 21 parameters of either the cell as a whole, or particular cellular structures. Maximum diameter (8.09 microns), perimeter (26.51 microns), area (36.87 microns2) and form factor (0.626) for the cell as a whole, and maximum diameter (5.08 microns), perimeter (18.74 microns), area (12.54 microns2) and form factor (0.452) for the nucleus were determined. Also measured were nuclear-cytoplasmic ratio (0.5595), filament thickness (10 nm), and granular numerical density (7.02 granules/micron2). Maximum diameter, area, and form factor of neurosecretory granules were 94.23 nm, 5020.05 nm2, and 0.93, respectively. Length of desmosomes linking NEMCs to keratinocytes was determined (286.9 nm) and compared with that of interkeratinocytic desmosomes (385 nm). In addition, length and diameter of cellular processes (spine-like processes (1.58 micron X 0.26 micron), interstitial processes (1.39 micron X 0.25 micron), and microvilli (0.35 micron X 0.25 micron) were measured after separation and classification according to the particular morphologic characteristics of each.     

The "colloid body"--its nature and pathogenesis.

The so-called “colloid bodies” in lichen planus and lichen amyloidosus were observed ultrastructurally and compared with the changes of basal cells in “Futorafur dermatitis”, which occurs after long term administration of N₁-(2′-tetrahydrofuryl-5-fluorouracil). The “colloid bodies” were located in the intercellular spaces in the epidermis and freely in the stroma of the upper corium. They were mainly composed of fine filaments, and frequently contained some melanosomes, mitochondria and other vacuolated structures. These filaments had a constant width about 70 to 80 Å in diameter, which was much the same as those of tonofilaments in an adjacent keratinocytes. A number of desmosomes, to which these filaments convergently adhered, were occasionally observed in “colloid bodies”. This finding seems to support the theory that these filaments are themselves tonofilaments originating from keratinocytes, and they are fundamentally differentiated from other types of filaments in the dermo-epidermal junction areas. The degenerative processes of basal cells in the epidermis in the lesions of “Futorafur dermatitis” seemed to be identical to those in formation of “colloid bodies” from keratinocytes in lichen planus.     

Nanostructure of the epidermal extracellular space as observed by cryo-electron microscopy of vitreous sections of human skin

The newly developed method, cryo-electron microscopy of vitreous sections, was used to observe the nanostructure of the epidermal extracellular space. The data were obtained from vitreous sections of freshly taken, fully hydrated, non-cryo-protected human skin. The extracellular space of viable epidermis contains desmosomes, expressing a characteristic extracellular transverse approximately 5 nm periodicity, interconnected by a relatively electron lucent inter-desmosomal space. The extracellular space between viable and cornified epidermis contains transition desmosomes at different stages of reorganization interconnected by widened areas expressing a rich variety of complex membrane-like structures. The extracellular space of cornified epidermis contains approximately 9, approximately 14, approximately 25, approximately 33, approximately 39, approximately 44, and approximately 48 nm thick regions in turn containing one, two, four, six, eight, eight, and ten parallel electron-dense lines, respectively, between adjacent corneocyte lipid envelopes. The eight-line approximately 44 nm thick regions are most prevalent.     

Decreased retinyl ester concentrations in UV-induced murine squamous cell carcinomas

Squamous cell carcinomas were induced in hairless mice by repeated irradiations with UVB (280-320 nm, total dose 30 J/cm2) plus UVA (320-400 nm, total dose 168 J/cm2). The irradiated animals and non-irradiated controls were fed on diets with or without vitamin A supplementation (20,000 IU/kg). At the appearance of tumours, 30 to 43 weeks after the last irradiation, the vitamin A (retinol plus retinyl ester) concentrations in the serum, liver, epidermis and tumours and the retinol esterifying activities in microsomes from epidermis and tumours were measured. The liver and epidermal vitamin A concentrations were 2-3 times higher in vitamin A supplemented than in unsupplemented animals, but did not differ between tumour-bearing animals and non-irradiated controls receiving identical diets. The vitamin A concentration in the tumours was significantly lower than in perilesional epidermis. The largest difference (p less than 0.001) between the tumour and epidermal values was observed in the vitamin A supplemented group. The low vitamin A content of the tumours was entirely due to a marked (2 to 6-fold) reduction in the retinyl ester fraction. In contrast, the retinol content of the tumours was increased to twice that of normal epidermis. The activity of the esterifying enzyme, acyl-CoA:retinol acyltransferase (EC 2.3.1.76), was unchanged. The reason for the reduced retinyl ester concentration thus remains unclear. Still, it is possible that a disturbed interconversion of retinol to retinyl esters plays a role in murine photo-carcinogenesis.     

Desmocollin 1 and desmoglein 1 expression in human epidermis and keratinizing oral mucosa: a comparative immunohistochemical and molecular study

Epidermis and keratinizing oral mucosa (KOM) are effective barriers against a wide spectrum of insults. The optimal form of protection provided by each epithelium is determined also by the molecular composition of desmosomes. Up to now, the expression of the “skin type” desmosomal cadherins, i.e. desmocollin 1 (Dsc1) and desmoglein 1 (Dsg1), was correlated with the morphological features of keratinocyte terminal differentiation in epidermis, but not in KOM. The aim of the present study was to investigate Dsc1 and Dsg1 expression in adult human KOM compared to epidermis. Biopsies of epidermis and KOM were obtained from young healthy adults (n=6) and simultaneously processed for immunofluorescence analysis, post-embedding immunogold electron microscopy (immunogold EM), and RT-PCR analysis. For molecular biology analysis, as a negative control, we considered human fibroblasts. By immunofluorescence and immunogold EM, Dsc1 labeling was not detected in any suprabasal layer of KOM, but it was present in the upper spinous/granular layers of epidermis. Immunofluorescence and transmission electron microscopy analysis showed that (i) Dsg1 expression was evident in the spinous, granular, and horny layer of the oral epithelium and (ii) Dsg1 immunoreactivity was always lower in desmosomes between oral keratinocytes than in all epidermal junctions. RT-PCR analysis confirmed that in KOM Dsc1 gene expression was undetectable. On the whole, these observations suggest a weakened adhesion in KOM, allowing oral keratinocytes to undergo a faster transition throughout the living layers of the epithelium. The intrinsic and specific regulation of the molecular composition of desmosomes can contribute in defining a specific keratinocyte phenotype in KOM and in epidermis.     

Epidermolysis bullosa herpetiformis (Dowling-Meara type) exhibits ultrastructural derangement of tonofilaments and desmosomes

Ultrastructural and immunohistochemical studies of clinically intact skin obtained from three severe neonatal cases of epidermolysis bullosa herpetiformis (Dowling-Meara type) demonstrated disorders in the assembly of keratin intermediate filaments and desmosomes of the keratinocytes. During mitosis, K5- and K14-positive and K1- and K10-negative tonofilaments were disrupted and formed spherical bodies associated with intracytoplasmic desmosomes by invagination of the desmosomes and the adjacent plasma membrane. During the invagination process, destructive changes in the internalized membrane were noted. These were accompanied by gradual loss of reactivity with a monoclonal antibody ZK31, which detected plasma membrane adjacent to the attachment plaques of desmosomes. However, the reactivity of the attachment plaques of the internalized desmosomes for desmoplakins and desmoglein did not decline during the process of internalization. In the suprabasal layers of the epidermis, filamentous substructures and K1 and K10 appeared at the periphery of the spherical bodies. Simultaneously, the desmosomes that were sparsely located in the lower epidermis, increased in number as cell differentiation progressed. Thus, the keratinocytes attained an almost normal appearance with respect to tonofilaments and desmosomes by the time they reached the upper layer of the epidermis. These findings may be relevant to the mechanism responsible for the clinical appearance of the herpetiform blisters in epidermolysis bullosa herpetiformis, which are also characterized by spontaneous involution during childhood or when exposed to high ambient temperatures.Part of this work was presented at the annual meeting of the Japanese Society for Ultrastructural Cutaneous Biology, 12 May 1990, Tokushima, Japan, at the joint meeting of the Society for Cutaneous Ultrastructure Research and the Japanese Society for Ultrastructural Cutaneous Biology, 23–25 May 1991, Vienna, Austria, and at the sixth Conference on Disorders of Keratinization, 6 July 1991, Tokyo, Japan     

Immunochemical analysis of the distribution of the desmosomal protein desmoglein I in different layers of plantar epidermis

An antiserum raised against the bovine desmosomal protein desmoglein I (DGI), Mr approximately 160 kDa, was used in an immunochemical analysis of human plantar epidermis. Different layers of the tissue were prepared by means of horizontal freeze sectioning. Loosely attached surface layers were obtained by means of scraping of the skin surface with a scalpel. Tissue extracts were analysed by means of sodium dodecylsulphate polyacrylamide gel electrophoresis followed by immunoblotting. Significant amounts of a component with Mr approximately 160 kDa, reactive with the DG I-antiserum, were found in all layers except the loosely attached surface layers. In these layers the antiserum detected a component with Mr approximately 80 kDa, not found in other layers. This component may be a degradation product of DG I. Since DG I belongs to the group of transmembrane desmosomal proteins that is believed to constitute the link between the intracellular parts of desmosomes of opposing cells, it is concluded that desmosomes may play an important role in plantar stratum corneum cell cohesion, and that degradation of desmosomes may be an important step in desquamation in plantar epidermis.     

Measurement of erythemal response to ultraviolet radiation by “monochromatic” photography

Summary To investigate the turnover of complete junctional membrane areas (domain turnover), psoriatic epidermis, keratoacanthoma, and squamous cell carcinoma were studied with an electron microscope and compared with the normal epidermis.Three types of domain turnover of junctions were observed. First, the uptake of intact intercellular junctions, which occurred as complete or incomplete annular junctions in the cytoplasm. Mainly gap junctions were involved. Second, the formation of autojunctions bridging an infolding at the surface of a cell, which is then incorporated and occurs partly with an attached saccule in the cytoplasm. This type was confined to desmosomes. Finally, the loss of intercellular desmosomes into the intercellular space was observed.Proliferating epidermis was associated with an increased domain turnover of gap junctional membrane areas. Domain turnover of desmosomes prevailed in tumors.Supported by the Deutsche Forschungsgemeinschaft (Ma 674/3-2). Presented at the 8th Annual Meeting of the Society for Cutaneous Ultrastructure Research (SCUR), London, May 21–23, 1981     

The growth and differentiation of cultured newborn rat keratinocytes

Keratinocytes were cultured from adult and newborn rat epidermis using the 3T3 feeder cell technique. By modifying culture conditions a long-lived line of newborn rat keratinocytes was developed which showed a plating efficiency of 40% and a doubling time of 16 h. The cells produced stratified colonies with tonofilaments, desmosomes, cell envelopes, and keratohyaline granules. When the cells were grown on a collagen gel they formed a thick stratum corneum and many keratohyaline granules. The fibrous proteins synthesized by the newborn rat cultured keratinocytes were different than those of newborn rat epidermis but similar to those of adult rat cultured keratinocytes. A histidine-rich basic protein was identified by immunologic techniques but it appeared to be more heterogeneous than that of newborn rat epidermis. A cell envelope precursor protein was identified by dansyl cadaverine incorporation studies and was identical to a major envelope precursor of newborn rat epidermis. The growth characteristics, colony morphology, and biochemical markers did not change for up to 40 passages and there was no evidence of malignant transformation. Because of their case of growth and long-term survival these cells are useful for studying a variety of problems related to keratinization.     

Epidermal Langerhans' cells in Granuloma Annulare

Langerhans' cells were studied in the epidermis of two patients with Granuloma Annulare (GA) of the generalized type and compared with those in two normal controls. The percentage of Langerhans' cells in the GA epidermis population was 3.4% and in the control, 1.8%. The density of Langerhans' cells in different adjacent sites of the same epidermis was not uniform, being in the range of 0.9–5% in GA and 0.6–4% in controls. In the GA epidermis, the Langerhans' cell section area was increased by about 40% and the number of Langerhans' cell granules found per cell section by about 41%. The findings may indicate that, in granuloma annulare epidermis, the Langerhans' cells are in a more active state.     

Ultrastructural studies of acantholysis induced in vivo by passive transfer of IgG from endemic pemphigus foliaceus (Fogo Selvagem)

Intraperitoneal (IP) injections of IgG from patients with Endemic Pemphigus Foliaceus [Fogo Selvagem (FS)] cause acantholysis in BALB/c mice (JID. 85:538, 1985). The dynamic ultrastructural changes of FS IgG-induced acantholysis in mice are the subject of this study. FS IgG was injected IP into neonatal BALB/c mice. Skin and serum was studied at 0, 1, 3, 6, 12, 18, and 24 h post injection by immunofluorescence (IF), electron microscopy (EM), and immuno-EM. Binding of FS IgG in the intercellular spaces (ICS) of the basal cell layer was seen by IF within 1 h and was strongest at 12 h. IgG binding affected the spinous and granular cell layer by 12 h, then faded and remain localized only in the basal cell layer at 24 h. By immuno-EM, IgG binding was diffuse along the keratinocyte surface. Edema of the ICS in the basal cell layer was present at 1 h by EM. At 12 h, there was microvillous formation with intact desmosomes at the tip of the projections. Splitting of desmosomes (forming half desmosomes) and acantholysis primarily affecting the granular cell layer were most prominent between 12 and 24 h. The plaques of the half desmosomes gradually disappeared and tonofilaments retracted into the cytoplasm. Detaching keratinocytes showed vacuolization, swollen mitochondria, and internalization of intact desmosomes and half desmosomes (remnants of split desmosomes). This investigation shows that the ultrastructural changes observed in the epidermis of patients with FS can be duplicated in experimental animals by IP injection of FS IgG. Further, FS IgG may have direct effects on the assembly/disassembly of desmosomes.     

干细胞因子及其受体在寻常型白癜风表皮中的表达

目的 探讨SCF/c-kit信号通路在白癜风发病中的作用。方法 采用免疫组化和RT-PCR法检测17例寻常型稳定期白癜风患者和10例正常对照标本中表皮角质形成细胞的干细胞因子表达及基底层黑素细胞c-kit的表达情况。结果 白癜风非皮损区干细胞因子、c-kit蛋白表达与正常对照无明显差异(P>0.05),皮损区干细胞因子表达显著高于正常对照皮肤(P<0.05),而c-kit表达显著低于正常对照皮肤(P<0.05)。白癜风非皮损区表皮干细胞因子、c-kit mRNA表达平均水平与正常对照近似(P>0.05);皮损区干细胞因子mRNA表达水平高于非皮损区及正常对照组差异有统计学意义(P<0.05);皮损区c-kit mRNA表达水平显著低于非皮损区及正常对照组(P<0.05)。结论 SCF/c-kit的异常表达可能与白癜风的发病有关。     

Trichilemmal keratosis (horn): a light and electron microscopic study

Three cases of trichilemmal keratosis (horn) were light microscopically examined and all showed numbers of U- or V-shaped epidermal proliferations which keratinized in a fashion either identical or similar to trichilemmal keratinization. Electron microscopy revealed both uneven and linear borders between the keratinized and the keratinizing cells with a few keratohyalin droplets, remnants of desmosomes, no marginal band in the horny layer, perinuclear vacuolation, few spherical bodies in the intercellular spaces (ICS) of the upper epidermis, and widening of the ICS of the lower epidermis. A number of electron dense spherical particles, 40-50 nm in diameter, were observed in nuclei of the upper epidermis. This suggests that ultrastructure of trichilemmal keratosis is similar rather to viral warts than to trichilemmal cysts, although there are close similarities between trichilemmal keratosis and cyst.     

Immunomolecular mapping of adherens junction and desmosomal components in normal human epidermis

Adherens junctions (AJs) are cell-cell and cell-matrix junctions that are known to comprise the transmembrane and cytoplasmic components linked to the f-actin cytoskeleton. Although the presence of AJs han been confirmed in normal human epidermis, previous studies immunolocalizing AJ-related antigens have been controversial. The purpose of this study was to produce a more precise molecular mapping of AJs and their constituents in relation to desmosomes in normal human epidermal keratinocytes. Using an electron microscope (EM) method to optimally fix plasma membranes. AJ structures were typically seen as a narrowing of the intercellular space between two keratinocytes that was distinct from desmosomes and gap junctions. Such structures were consistently found more frequently in the upper epidermis than in the basal layer. Immunogold electron microscopy showed an absence of the AJ components (E-cadherin and beta-catenin) from desmosomal areas but they were present at interdesmosomal areas at sites of close membrane association. Conversely, the desmosomal components plakoglobin and plakophilin 1 were restricted only to the outer attachment plaque of the desmosome. These results further confirm that AJs have a distinct molecular composition and distribution from desmosomes and that they regularly occur between desmosomes along the keratinocyte plasma membrane to provide alternative cell-cell adhesion mechanisms.