Human epidermal melanocyte and keratinocyte melanocortin receptors: Visualization by melanotropic peptide conjugated microspheres (latex beads)

Jinwen Jiang, Shubh D. Sharma, Victor J Hruby, David L. Bentley, Jody L. Fink, Mac E. Hadley

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

The objectives of this research were to determine whether melanocortin receptors are characteristic (constant) membrane markers of human epidermal melanocytes. Methodologies were developed to visualize melanotropin receptors by scanning electron microscopy (SEM). Multiple copies (up to a hundred) of [Nle4,D-Phe7]α-MSH, a superpotent analog of α-melanocyte stimulating hormone (α-MSH), were conjugated to a macromolecular carrier (latex beads: microspheres). Incubation in the presence of the melanotropin-conjugated microspheres resulted in binding of human normal epidermal melanocytes to the beads. Almost every (possibly all) melanocyte possesses melanocortin receptors as visualized by SEM. Specificity of binding of the macromolecular conjugate was demonstrated by several studies: 1) Binding of melanocytes to the microspheres was specific since it could be blocked by prior incubation of the cells in the presence of the unconjugated hormone analog; 2) microspheres lacking bound ligand did not bind to the melanocytes; 3) microspheres that were first treated with reducing agents (e.g., dithiothreitol) did not subsequently bind to melanocytes; 4) another peptide hormone ligand (e.g., a substance-P analog) attached to the latex beads failed to bind to the cells; 5) B16/F10 mouse melanoma cells known to express melanocortin receptors bound to the microspheres; and 6) cells of nonmelanocyte origin (e.g., mammary cancer cells, small-cell lung cancer cells, fibroblasts) did not bind to the macromolecular conjugate. One exception was that human epidermal keratinocytes also expressed melanocortin receptors as determined by all the criteria established above for epidermal melanocytes. Thus, cell specific melanocortin receptors appear to be characteristic cell surface markers of epidermal melanocytes and keratinocytes.

Original languageEnglish (US)
Pages (from-to)240-247
Number of pages8
JournalPigment Cell Research
Volume9
Issue number5
StatePublished - Oct 1996

Fingerprint

Receptor, Melanocortin, Type 1
Melanocortin Receptors
melanocytes
keratinocytes
Latex
latex
Microspheres
Keratinocytes
Melanocytes
Visualization
peptides
Melanocyte-Stimulating Hormones
Peptides
receptors
Cells
melanocyte-stimulating hormone
cells
Electron Scanning Microscopy
Ligands
Scanning electron microscopy

Keywords

  • Keratinocytes
  • Melanocortin receptors
  • Melanocyte stimulating hormone
  • Melanoma
  • MSH

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Plant Science
  • Cell Biology
  • Clinical Biochemistry
  • Developmental Biology

Cite this

Human epidermal melanocyte and keratinocyte melanocortin receptors : Visualization by melanotropic peptide conjugated microspheres (latex beads). / Jiang, Jinwen; Sharma, Shubh D.; Hruby, Victor J; Bentley, David L.; Fink, Jody L.; Hadley, Mac E.

In: Pigment Cell Research, Vol. 9, No. 5, 10.1996, p. 240-247.

Research output: Contribution to journalArticle

Jiang, Jinwen ; Sharma, Shubh D. ; Hruby, Victor J ; Bentley, David L. ; Fink, Jody L. ; Hadley, Mac E. / Human epidermal melanocyte and keratinocyte melanocortin receptors : Visualization by melanotropic peptide conjugated microspheres (latex beads). In: Pigment Cell Research. 1996 ; Vol. 9, No. 5. pp. 240-247.
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abstract = "The objectives of this research were to determine whether melanocortin receptors are characteristic (constant) membrane markers of human epidermal melanocytes. Methodologies were developed to visualize melanotropin receptors by scanning electron microscopy (SEM). Multiple copies (up to a hundred) of [Nle4,D-Phe7]α-MSH, a superpotent analog of α-melanocyte stimulating hormone (α-MSH), were conjugated to a macromolecular carrier (latex beads: microspheres). Incubation in the presence of the melanotropin-conjugated microspheres resulted in binding of human normal epidermal melanocytes to the beads. Almost every (possibly all) melanocyte possesses melanocortin receptors as visualized by SEM. Specificity of binding of the macromolecular conjugate was demonstrated by several studies: 1) Binding of melanocytes to the microspheres was specific since it could be blocked by prior incubation of the cells in the presence of the unconjugated hormone analog; 2) microspheres lacking bound ligand did not bind to the melanocytes; 3) microspheres that were first treated with reducing agents (e.g., dithiothreitol) did not subsequently bind to melanocytes; 4) another peptide hormone ligand (e.g., a substance-P analog) attached to the latex beads failed to bind to the cells; 5) B16/F10 mouse melanoma cells known to express melanocortin receptors bound to the microspheres; and 6) cells of nonmelanocyte origin (e.g., mammary cancer cells, small-cell lung cancer cells, fibroblasts) did not bind to the macromolecular conjugate. One exception was that human epidermal keratinocytes also expressed melanocortin receptors as determined by all the criteria established above for epidermal melanocytes. Thus, cell specific melanocortin receptors appear to be characteristic cell surface markers of epidermal melanocytes and keratinocytes.",
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AB - The objectives of this research were to determine whether melanocortin receptors are characteristic (constant) membrane markers of human epidermal melanocytes. Methodologies were developed to visualize melanotropin receptors by scanning electron microscopy (SEM). Multiple copies (up to a hundred) of [Nle4,D-Phe7]α-MSH, a superpotent analog of α-melanocyte stimulating hormone (α-MSH), were conjugated to a macromolecular carrier (latex beads: microspheres). Incubation in the presence of the melanotropin-conjugated microspheres resulted in binding of human normal epidermal melanocytes to the beads. Almost every (possibly all) melanocyte possesses melanocortin receptors as visualized by SEM. Specificity of binding of the macromolecular conjugate was demonstrated by several studies: 1) Binding of melanocytes to the microspheres was specific since it could be blocked by prior incubation of the cells in the presence of the unconjugated hormone analog; 2) microspheres lacking bound ligand did not bind to the melanocytes; 3) microspheres that were first treated with reducing agents (e.g., dithiothreitol) did not subsequently bind to melanocytes; 4) another peptide hormone ligand (e.g., a substance-P analog) attached to the latex beads failed to bind to the cells; 5) B16/F10 mouse melanoma cells known to express melanocortin receptors bound to the microspheres; and 6) cells of nonmelanocyte origin (e.g., mammary cancer cells, small-cell lung cancer cells, fibroblasts) did not bind to the macromolecular conjugate. One exception was that human epidermal keratinocytes also expressed melanocortin receptors as determined by all the criteria established above for epidermal melanocytes. Thus, cell specific melanocortin receptors appear to be characteristic cell surface markers of epidermal melanocytes and keratinocytes.

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