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Very few histologic reports describe normal melanocytes of the nail unit. Previous studies predominantly address the distal nail matrix melanocytes; we found no review of nail-bed melanocytes in the literature. The proximal nail matrix melanocytes are difficult to identify; the cells cannot be identified by L-DOPA staining. More recently, their scarcity was confirmed by immunohistochemistry with a large panel of antibodies directed against melanocytes. We wished to detect the proximal nail matrix dormant melanocytes and compare their density and distribution with that of the other melanocytes in the distal matrix and nail bed and to establish criteria of normality that may help clarify the pathologic features of benign nevoid melanonychia in the nails of whites. A panel of five monoclonal antibodies (MoAbs), including HMB45 and TRP1 directed against antigens localized in early melanosomal vesicles, was investigated in frozen sections of six nail specimens from whites. Both vertical and horizontal sections were assessed to determine the presence of dormant melanocytes. Results showed that the proximal nail matrix melanocytes were clearly identified with MoAbs HMB45 and tyrosinase-related protein-1 (TRP-1). By contrast, melanocytes stained by MoAb against tyrosinase and L-DOPA reaction were evident, especially in the distal matrix. With MoAb TRP-1, the epithelial sheets showed counts of approximately 217+/-84/mm2 in the proximal matrix and of 132+/-34/mm2 in the distal matrix; the nail bed counts were only 45+/-25/mm2. The split epithelial sheets had 103+/-17/mm2 L-DOPA-positive melanocytes in the distal third of the matrix, but only a few of them were detected in the proximal matrix and none were noted in the nail bed. We clearly identified proximal nail melanocytes using MoAb HMB45 and TRP1. The total number of matrix melanocytes can be estimated as approximately 217/mm2. In proximal matrix, the dormant melanocytes compartment was predominant. In the distal matrix, two compartments were identified: a functionally differentiated and a dormant compartment. Contrary to classical opinion, longitudinal melanonychia originates more frequently in the distal matrix, not secondary to the larger melanocyte density but because only the distal matrix contains an active melanin synthesis compartment. Furthermore, the superficial distribution of proximal nail melanocytes in vertical sections showed a histologic feature that may simulate the pagetoid pattern of melanoma in situ.

The distribution of Merkel cells (MCs) in normal human skin and mucosa was studied using the mouse monoclonal antibody Troma-1, reacting specifically with component 8 of the Moll cytokeratin catalogue. The specificity of this antibody for MCs in human skin was assessed by double indirect immunofluorescence (IIF) and immunoelectron microscopy. Two-hundred and thirty 6-mm punch biopsies were obtained from 44 different sites from six human cadavers within 48 h post-mortem. IIF was performed with Troma-1 on EDTA-split epithelial sheets and the MCs were counted and the mean values per mm2 calculated for each site. Regions with greater than 50 MC/mm2 were the lips, hard palate, palms, finger pads, proximal nail fold, and dorsum of the feet. Three different patterns were observed in the epidermis or mucosa: MCs grouped in clumps, linear and arciform arrangements, and scattered MCs. In the hair follicles grouped MCs were observed in the bulb and scattered MCs were seen in the outer root sheath.

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A fully differentiated epithelium displaying features of human epidermis was obtained in vitro by culturing second-passage normal human keratinocytes for 14 days in defined medium and on an inert polycarbonate filter substratum at the air-liquid interface. Vertical sections stained for histology and indirect immunofluorescence studies showed that the 'basal' cells synthesize and secrete all major markers of hemidesmosomes and the lamina lucida. Components of the lamina densa are also expressed. Collagen VII is synthesized, but not secreted. Ultrastructural studies showed the presence of hemidesmosomes with major dense plaques and anchoring filaments, and a basement membrane-like structure was clearly identified. These results show that epidermal cells are able to produce hemidesmosomes and to secrete the major components of the dermo-epidermal junction in the absence of serum and dermal factors, suggesting that basement membrane synthesis and hemidesmosome assembly are not dependent on the presence of dermis.