Skip to main content
Log in

The appearance, density and distribution of melanocytes in human embryonic and fetal skin revealed by the anti-melanoma monoclonal antibody, HMB-45

  • Published:
Anatomy and Embryology Aims and scope Submit manuscript

Summary

The presence, densities, and patterns of distribution of melanocytes in the epidermis of human embryos and fetuses, ranging in age from 40 d to 140 d estimated gestational age (EGA), were studied using the HMB-45 monoclonal antibody that recognizes an antigen in melanoma cells and fetal melanocytes. Immunostained sections of skin and epidermal sheets revealed dendritic melanocytes within the basal or intermediate layers of 50 d EGA and older skin. Melanocytes could not be identified by immunostaining or electron microscopy in younger (40–50 d EGA) epidermis or in cultured epidermal cells from these specimens. However, skin from a 45 d EGA embryo grown in organ culture for 11 d stained positively with HMB-45, suggesting that melanocytes are present at that age either in the epidermis or dermis of the explant. Double-labeling experiments using ATPase and HMB-45 confirmed the specificity of HMB-45 for melanocytes and demonstrated that melanocytes and Langerhans cells are nonoverlapping populations. Melanocytes were present in the embryonic epidermis in relatively high numbers (mean value of ∼1050 cells/mm2); they increased in density to ∼2300 cells/mm2 during the late first trimester and early second trimester, then declined during later stages of development to a density of ∼800 cells/mm2, within the range of values for the newborn child and young adult. Equivalent numbers of melanocytes were recognized by silver staining and with the HMB-45 antibody in an 87 d EGA test sample, indicating that HMB-45 reacted with the total melanocytic population. Melanocytes appeared to be distributed in epidermal sheets in a regular pattern. Statistical tests used to evaluate the randomness of a population revealed a tendency toward a non-random distribution in specimens younger than 80 d EGA, just prior to appendage formation and epidermal stratification into multiple layers, however there was variability in the degree of randomness for any given age. The results of this study have closed the gap in timing between the conclusion of neural crest formation and migration (around 6 weeks) and the appearance of melanocytes in the skin between 40–50 d EGA.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Aherne WA, Diggle PJ (1978) The estimation of neuronal population density by a robust distance analysis. J Microsc 114:285–293

    Google Scholar 

  • Aherne WA, Dunnill MS (1982) Morphometry, Edward Arnold, London, pp 137–141

    Google Scholar 

  • Albino A, Houghton AN (1985) Cell surface antigens of melanocytes and melanoma. Cancer Surv 4:185–211

    Google Scholar 

  • Adachi B (1903) Hautpigment beim Menschen und bei den Affen. Z Morphol Anthropol 6:1–131

    Google Scholar 

  • Baelz E (1885) Die körperlichen Eigenschaften der Japaner. Mitteilung Deutsche Gesellschaft für Natur und Völkerkunde Ostasiens 4:40

    Google Scholar 

  • Barry J (1953) Recherches sur l'origine des cellulares de Langerhans de l'epiderme. Biomed Pharmacother 42:293–315

    Google Scholar 

  • Barthelmez GW, Evans HM (1926) Development of the human embryo during the period of somite formation; including embryos with 2 to 16 pairs of somites. Contrib Embryol Carnegie Inst 17:1–67

    Google Scholar 

  • Bartelmez GW, Blount MP (1954) The formation of neural crest from the primary optic vesicles in man. Contrib Embryol Carnegie Inst 35:5–71

    Google Scholar 

  • Becker Jr SW, Zimmerman AA (1955) Further studies on melanocytes and melanogenesis in the human fetus and newborn. J Invest Dermatol 25:103–112

    Google Scholar 

  • Besag JE, Gleaves JT (1973) On the detection of spatial pattern in plant communities. Bull Int Inst Statist 45:153–158

    Google Scholar 

  • Bickenbach JR, Holbrook KA (1987) Label-retaining cells in human embryonic and fetal epidermis. J Invest Dermatol 88:42–46

    Google Scholar 

  • Bloch B (1921) Über die Entwicklung des Haut- und Haarpigmentes beim menschlichen Embryo und über das Erlöschen der Pigmentbildung im ergrauenden Haar. Arch Dermatol Syph 135:77–108

    Google Scholar 

  • Breathnach AS (1971) An atlas of the ultrastructure of human skin. Churchill, London, pp 1–398

    Google Scholar 

  • Breathnach AS, Robins J (1969) Ultrastructural features of epidermis of a 14 mm (6 weeks) human embryo. Br J Dermatol 81:504–516

    Google Scholar 

  • Breathnach AS, Wyllie LM (1965) Electron microscopy of melanocytes and Langerhans cells in human fetal epidermis at fourteen weeks. J Invest Dermatol 44:51–50

    Google Scholar 

  • Bronner-Fraser M (1987) Perturbation of cranial neural crest migration by the HNK-1 antibody. Dev Biol 123:321–331

    Google Scholar 

  • Diggle PJ (1977) The detection of random heterogeneity in plant populations. Biometrics 33:390–394

    Google Scholar 

  • Ehrmann S (1885) Untersuchungen über die Physiologie und Pathologie des Hautpigmentes. Arch Dermatol Syph 17:507–532

    Google Scholar 

  • El Bahrawy AA (1922) Über den Mongolenfleck bei Europaern. Ein Beitrag zur Pigmentlehre. Arch Dermatol Syph 141:171–192

    Google Scholar 

  • Esclamado R, Gown AM, Voegl AM (1986) Unique proteins defined by monoclonal antibodies specific for human melanoma. Some potential clinical applications. Am J Surg 152:376–385

    Google Scholar 

  • Foster CA, Holbrook KA, Farr AG (1986) Ontogeny of Langerhans cells in human embryonic and fetal skin: Expression of HLA-DR and OKT6 determinants. J Invest Dermatol 86:240–243

    Google Scholar 

  • Foster CA, Holbrook KA (1989) Ontogeny of Langerhans cells in human embryonic and fetal skin: cell densities and phenotypic expression relative to epidermal growth. Am J Anat 184:157–164

    Google Scholar 

  • Fujita H, Asgami C, Oda Y, Yamamoto K, Uchihara T (1970) Electron microscope study on the embryonic differentiation of the epidermis in human skin. Arch Histol Jpn 32:355–373

    Google Scholar 

  • Fukaya T, Hashimoto K, Eto H, Mehregan A (1986) Mouse monoclonal antibody (FKH1) detecting human melanoma-associated antigens. Cancer Res 46:5195–5200

    Google Scholar 

  • Gilchrest BA, Treloar V, Grassi AM, Yaar M, Szabo G, Flynn E (1986) Characteristics of cultivated adult human nevocellular nevus cells. J Invest Dermatol 87:102–107

    Google Scholar 

  • Gown AM, Vogel AM (1984) Monoclonal antibodies to human intermediate filament proteins: II. Distribution of filament proteins in normal human tissues. Am J Pathol 114:309–321

    Google Scholar 

  • Gown AM, Vogel AM, Hoak D, Gough F, McNutt MA (1986) Monclonal antibodies specific for melanocytic tumors distinguish subpopulations of melanocytes. Am J Pathol 123:195–203

    Google Scholar 

  • Grimm F (1895) Beiträge zum Studium des Pigments. Dermatol Z 2:328–343

    Google Scholar 

  • Hamada H (1972) Age changes in melanocyte distribution of the normal, human epidermis. Jpn J Dermatol 82:223–232

    Google Scholar 

  • Hashimoto K (1971) The ultrastructure of the skin of human embryos. VIII. Melanoblast and intrafollicular melanocyte. J Anat 108:99–108

    Google Scholar 

  • Holbrook KA (1983) Structure and function of the developing human skin. In: Goldsmith LA (ed) Biochemistry and Physiology of the Skin, vol 1. Oxford Univ Press, New York, 64–101

    Google Scholar 

  • Holbrook KA, Odland GF (1980) Regional development of the human epidermis in the first trimester embryo and the second trimester fetus ages related to the timing of amniocentesis and fetal biopsy. J Invest Dermatol 80:161–168

    Google Scholar 

  • Holbrook KA, Vogel AM, Foster CA, Underwood R (1987) Melanocytic cells in human embryonic and fetal epidermis as identified by the anti-melanoma monoclonal antibody, HMB-45. J Invest Dermatol 88:495A

    Google Scholar 

  • Holbrook KA, Vogel AM, Underwood RA, Foster CA (1988) Melanocytes in human embryonic and fetal skin: A review and new findings. Pig Cell Res 1 (Suppl):6–17

    Google Scholar 

  • Hsu S-M, Soban E (1982) Color modification of diaminobenzidine (DAB) precipitation by metallic ions and its application for double immunohistochemistry. J Histochem Cytochem 30:1079–1082

    Google Scholar 

  • Ishihawa N (1924) Über den sogenannten Mongolenfleck bei Japanischen foten. Folia Anat Japonica 2:1–4

    Google Scholar 

  • Ito M (1953) Studies on melanin. Tohoku J Exper Med 55 (Suppl 1)

  • Jimbow K, Quevedo WC, Fitzpatrick TB, Szabo G (1976) Some aspects of melanin biology: 1950–1975. J Invest Dermatol 67:72–89

    Google Scholar 

  • Karnvsky MJ (1965) A formaldehyde-glutaraldehyde fixative for use in electron microscopy. J Cell Biol (abstr) 27:137A

    Google Scholar 

  • Luft JH (1961) Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol 9:409–414

    Google Scholar 

  • Mierowsky E (1940) A critical review of pigment research in the last hundred years. Br J Dermatol 52:205–217

    Google Scholar 

  • Mishima Y, Widlan S (1966) Embryonic development of melanocytes in human hair and epidermis. J Invest Dermatol 46:263–277

    Google Scholar 

  • Morison D (1889) Beiträge zur Frage von der Pigmentbildung in the Negerhaut. Monatshefte prakt Dermatol 9:485–490

    Google Scholar 

  • Puchtler H, Waldrop FS, Meloan SN, Terry MS, Conner HM (1970) Methacarn (methanol-Carnoy) fixation: Practical and theoretical considerations. Histochemie 21:97–117

    Google Scholar 

  • Rawles ME (1947) Origin of pigment cells from the neural crest in the mouse embryo Physiol Zool 20:248–266

    Google Scholar 

  • Reynolds ES (1963) The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J Cell Biol 17:208–213

    Google Scholar 

  • Richardson KC, Jarett L, Finke EH (1960) Embedding in epoxy resins for ultrathin sectioning in electron microscopy. Stain Technol 35:313–323

    Google Scholar 

  • Sagebiel RW, Odland GF (1972) Ultrastructural identification of melanocytes in early human embryos. In: Riley V (ed) Pigmentation: Its Genesis and Biological Control. Appleton-Century-Croft, New York, pp 43–50

    Google Scholar 

  • Schutte B, Reynders MMJ, Bosman FT, Blijham GH (1987) Studies with anti-bromodeoxyuridine antibodies. II. Simultaneous immunocytochemical detection of antigen expression and DNA synthesis by in vivo labeling of mouse intestinal mucosa. J Histochem Cytochem 35:371–374

    Google Scholar 

  • Sikora LKJ, Demetrick DJ, Dixon WT, Uranski SJ, Temple W, Jerry LM (1987) Characterization of a novel neuroglandular antigen (NGA) expressed on abnormal human melanocytes. Int J Cancer 39:138–145

    Google Scholar 

  • Smith LT, Holbrook KA, Byers PH (1982) Structure of the dermal matrix during development and in the adult. J Invest Dermatol 79:93s-104s

    Google Scholar 

  • Smith LT, Holbrook KA, Madri JS (1986) Collagen types I, III and V in human embryonic and fetal skin. Am J Anat 175:507–522

    Google Scholar 

  • Snell RS, Bischitz PG (1963) The melanocytes and melanin in human abdominal wall skin: a survey made at different ages in both sexes and during pregnancy. J Anat 97:361–376

    Google Scholar 

  • Starrico RJ, Pinkus H (1957) Quantitative and qualitative data on the pigment cells of adult epidermis. J Invest Dermatol 28:33–44

    Google Scholar 

  • Szabo G (1969) Regional anatomy of human skin with special reference to the distribution of hair follicles, sweat glands and melanocytes. Philos Trans R Soc Lond (Biol Sci) 252:447–485

    Google Scholar 

  • Thomson A (1891) Note on the skin and scalp of the Negro fetus. J Anat Physiol 25:282–285

    Google Scholar 

  • Werkmeister JA, Triglia T, Mackay IR, Dowling JP, Varigos GA, Morstyn G, Burns GF (1987) Fluctuations in the expression of a glycolipid antigen associated with differentiation of melanoma cells monitored by a monclonal antibody, Leo Mel 3. Cancer Res 47:225–230

    Google Scholar 

  • Zimmerman AA (1950) The development of epidermal pigmentation in the Negro fetus. Proc. 2nd Conf. on Biology of Normal and Atypical Pigment Cell Growth. Zoologica 35:10–12

    Google Scholar 

  • Zimmerman AA, Becker SW, Jr (1959a) Melanoblasts and melanocytes in fetal Negro skin. Illinois Monographs in Medical Science, VI (3), Univ Illinois Press, Urbana, IL, pp 1–5

    Google Scholar 

  • Zimmerman AA, Becker SW, Jr (1959b) Precursors of epidermal melanocytes in the Negro Fetus. In: Gordon M (ed) Pigment Cell Biology, Academic Press, New York, pp 159–170

    Google Scholar 

  • Zimmerman AA, Cornbleet T (1948) The development of epidermal pigmentation in the Negro fetus. J Invest Dermatol 11:383–395

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Holbrook, K.A., Underwood, R.A., Vogel, A.M. et al. The appearance, density and distribution of melanocytes in human embryonic and fetal skin revealed by the anti-melanoma monoclonal antibody, HMB-45. Anat Embryol 180, 443–455 (1989). https://doi.org/10.1007/BF00305119

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00305119

Key words

Navigation