Human Merkel Cells Research Articles

Absence of Merkel cells in lesional skin of vitiligo [corrected

In vitiligo, other than loss of melanocytes and melanin pigment in the lesional skin, keratinocytes are also involved. Human fetal Merkel cells are now generally considered to be derived from the epidermis. To date, no observations on Merkel cells in lesional skin of active vitiligo have been reported. Merkel cells were identified by indirect immunofluorescence microscopy using the monoclonal antibody TROMA 1. No TROMA 1-positive cells were observed in the vitiliginous skin lesions in any of the specimens examined, whereas normal numbers of these cells were seen in the adjacent pigmented skin. This finding suggests that the processes that lead to damage of keratinocytes in vitiligo also involve other keratin-expressing cells such as the Merkel cells.

Merkel cells in human fetal eccrine glands.

The presence of human Merkel cells in the eccrine ridges and eccrine germs was studied, using antibodies to simple epithelial keratins, in separated epidermal sheets with attached eccrine ducts. The localization of Merkel cells could be analysed three-dimensionally in the wet, whole-mount of the stained sheets. In the plantar skin of a 12-week-old human fetus, immunoreactive (ir-) Merkel cells were randomly located in the flattened epidermis. In the plantar skin of a 15-week-old human fetus, there was early development of eccrine germs, and Merkel cells were concentrated in eccrine gland ridges. In the plantar skin of a 20-week-old human fetus, eccrine germs were well formed and ir-Merkel cells were located within the developing eccrine ridges and ducts. In the plantar skin of adults, the eccrine concentration of Merkel cells was markedly reduced. Concentration of Merkel cells on the eccrine structures was also observed in the scalp skin of human fetuses. This tendency continued into adult life, although there was a marked reduction in the total number of Merkel cells. These findings suggest that epidermal Merkel cells move down into the eccrine ducts as eccrine germs extend into the mesenchyme. Alternatively, they may develop de novo from the keratinocytes of the eccrine duct. In view of the expression of nerve growth factor receptor in fetal Merkel cells, it is postulated that these eccrine gland Merkel cells play a role in the formation of the periglandular nerve plexus.

Early development of human Merkel cells.

Human fetal Merkel cells are now generally considered to be epidermal derivatives. Previous studies using antibodies against the simple epithelial cytokeratins (CKs), 8 and 18, have demonstrated the presence of these cells in the epidermis at as early as fetal week 10 to 12. Using antibodies against CK 20 whose expression within the skin is restricted to Merkel cells, we applied immunofluorescence and immunoperoxidase microscopy to analyze earlier embryonic and fetal human skin (wk 7 to 9). We were able to demonstrate the first Merkel cells at as early as fetal wk 8, i.e., at the same time as the epidermis starts to develop an intermediate, third layer, characterized by the expression of CKs 1, 10, and 11. Most of these early Merkel cells were localized above the basal layer. Their shape was round to oval, dendrites being infrequent and short. At fetal wk 9, Merkel cells were considerably more numerous. These results persuasively argue for a much earlier fetal development of Merkel cells within the epidermis than previously thought. A hypothesis concerning the differentiation of Merkel cells from embryonic basal keratinocytes is discussed.

Intraepidermal Formation of Merkel Cells in Xenografts of Human Fetal Skin

An experimental transplantation model using human fetal skin was applied to approach the question of the embryologic origin of human Merkel cells. Palmar and plantar skin from five fetuses, between 8 and 11 weeks of estimated gestational age (EGA), was xenografted to subcutaneous beds of nude mice. After 4 or 8 weeks of growth, biopsies were taken from these xenografts and examined for the presence of Merkel cells, using immunocytochemistry with antibodies specific for simple epithelial-type cytokeratins and neuron-specific enolase (NSE) as well as using electron microscopy. Skin from the same fetuses at the time of transplantation was screened in the same way. In all fetuses, no (or very scarce) epidermal Merkel cells were detected at the transplantation time, but in all cases abundant epidermal Merkel cells of apparent human origin were found after 4 or 8 weeks in xenograft culture. Dermal nerve fibers, as recognized by neurofilament antibodies, were scarce or essentially absent in the xenografts. These results indicate that Merkel cells regularly develop in epidermis dissected and xenografted in an early fetal stage, although the dissection implies the interruption of the dermal nerves. The results strongly support the notion of the origin of Merkel cells from epidermal precursor cells. The apparent absence of dermal Merkel cells and dermal nerve fibers in the xenografts suggests that the presence of dermal sensory nerve fibers may be required for the dropping off of epidermal Merkel cells into the upper dermis, which occurs in normal fetal development.

Calcitonin gene-related peptide (CGRP) immunoreactivity in the neuroendocrine Merkel cells and nerve fibres of pig and human skin.

The presence of calcitonin gene-related peptide (CGRP) in the skin of pig snout and human fingertip was investigated using immunohistochemical techniques. CGRP immunoreactivity was found in Merkel cells and nerve fibres of both species. In pig snout skin, Merkel cells containing CGRP were seen forming clusters at the tips of rete ridge epidermis and in the external root sheath of sinus hair follicles (vibrissae). Human Merkel cells immunostained for CGRP were found isolated or forming small groups in the basal layer of glandular epidermal ridges. In all cases, immunoreactivity was more intense on the side of the Merkel cell facing the associated nerve terminal (which was never positive for CGRP). This part of the Merkel cell has the greatest density of dense-cored granules, suggesting that CGRP must be stored in these granules. Nerve bundles containing CGRP-immunoreactive fibres were found at dermal and hypodermal level, and blood vessels were often surrounded by CGRP nerve fibres. In pig snout skin some nerve fibres containing CGRP penetrated the epidermis and terminated as free endings, and in the human fingertip a small number of CGRP-immunoreactive nerve fibres were seen in Meissner's corpuscles.

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.

Identification of Merkel cells in human skin by specific cytokeratin antibodies:: Changes of cell density and distribution in fetal and adult plantar epidermis

Merkel cells are special neurosecretory cells which, in adult human skin, are usually very scarce. By immunofluorescence microscopy using antibodies to human cytokeratin polypeptide no. 18, we localized distinct non-keratinocyte cells in the glandular ridges of human fetal and adult plantar epidermis. Using electron and immunofluorescence microscopy, these cells were identified as Merkel cells containing typical neurosecretory granules as well as bundles of intermediate-sized filaments and desmosomes. Two-dimensional gel electrophoresis of the cytoskeletal fractions of microdissected epidermal preparations highly enriched in Merkel cells indicated the presence of cytokeratin polypeptides nos. 8, 18 and 19 which are typical of diverse simple epithelia of the human body. Double immunofluorescence microscopy showed that these human Merkel cells contain neither neurofilaments nor vimentin filaments. In human fetuses of 18–24 weeks of age, conspicuously high concentrations of Merkel cells, reaching a density of approximately 1,700 Merkel cells/mm 2 skin, were found in the glandular ridges of plantar skin. The concentration decreased considerably at newborn and adult stages. Thin cell processes (up to 20 μm long) were observed in many fetal epidermal Merkel cells. In addition, we detected isolated Merkel cells deeper in the dermis (i.e. at distances of, at most, 100 μm from the epidermis) in fetal and newborn plantar skin. Our results show that Merkel cells are true epithelial cells which, however, differ profoundly from epidermal keratinocytes in their cytokeratin expression. The findings are discussed in relation to the much disputed question of the origin of Merkel cells. The present data speak against the immigration of Merkel cells from the neural crest, but rather suggest that they originate from epithelial cells of the skin, although most probably not from differentiated keratinocytes.