Three Modes of Melanosome Transfers in Caucasian Facial Skin: Hypothesis Based on an Ultrastructural Study

The transfer mechanism of melanosome from the melanocyte into the keratinocyte was investigated in mildly photodamaged Caucasian facial skin by electron microscopy. Three ways of transfer are suggested by our observations. The first mechanism probably occurs through the following process: 1) protrusion and insertion of the thick dendrite of the melanocyte into the basal keratinocyte, 2) formation of sac-dendrite complex in the subnuclear region, 3) digestion and segregation of the enclosed dendrite, 4) formation of the cistern in the paranuclear region, and 5) pinching-off of the melanosomes in single or aggregated form from the tip of the cistern. The second mechanism probably takes place through a membrane fusion between the melanocyte and the keratinocyte. Such a membrane fusion possibly forms a passage way for release of the melanosome from the former cell to the latter. The third mechanism is considered to include exocytosis of the single melanosome from the melanocyte followed by the endocytosis through the formation of coated-pit in the keratinocyte.     

In Situ and In Vitro Expression of Protein Kinase C Alpha in Human Melanocytes

Protein kinase C (PKC) is a multigene family of at least 12 isoforms involved in the transduction of extracellular signals. We investigated whether PKC-α, a major isoform known to be relatively abundant in brain tissue, is increased in human melanocytes relative to keratinocytes in vitro and in situ. Immunohistochemical staining for PKC-α in frozen neonatal human foreskin exhibited intermittent 2–3+ staining along the basal cell layer consistent with melanocytes, and 0–1+ staining of keratinocytes (on a scale of 0–3). Microscopic densitometry of the intermittent cellular staining was at least 3-fold greater than that of adjacent keratinocyte cell cytoplasm. Sequential frozen sections revealed similar intermittent cell staining with PKC-α and Mel-5 (tyrosinase related protein-1), known to specifically react with melanocytes. Northern blot analysis with a specific cDNA probe for PKC-α showed strong PKC-α mRNA expression in cultured melanocytes, whereas PKC-α mRNA in cultured non-stratifying keratinocytes was expressed at low levels. Western blot analysis revealed a prominent PKC-α band at approximately 80 kDa in melanocytes as opposed to a weak band in keratinocytes. Densitometry of the northern and western blots revealed that melanocytes had at least 10-fold more PKC-α mRNA and approximately 6-fold more PKC-α protein expression than keratinocytes. Total PKC activity measured in vitro revealed that melanocytes had 5-fold more activity than keratinocytes. The marked difference in melanocyte and keratinocyte expression of PKC-α provides further evidence for cell type specificity in the balance of PKC-α expression and may implicate differential PKC isoform signaling pathways in neuro-ectodermally derived cells.