Gossypol, a polyphenolic compound known to interfere with spermatogenesis, was found to have differential cytotoxic effects on normal and malignant melanocytes in culture. Ultrastructurally it caused marked swelling and vacuolization of mitochondria; there was an almost complete loss of cristae but the outer mitochondrial membrane was retained. The cytotoxicity seemed to be selective for actively proliferating cells, regardless of normal or malignant origin. An endothelial cell line (RF/6A) derived from the choroid-retina of the eye of a rhesus fetus was extremely sensitive to its toxic effect. The mechanism of action is still unknown. The fact that mitochondria are predominantly damaged suggests that toxicity involves a perturbation of energy metabolism and the membrane transport system.

We have shown that dicarboxylic acids (C9 and C12), known competitive inhibitors of tyrosinase, are selectively cytotoxic to malignant melanogenic melanocytes but not to normal pigmented cells or to amelanotic or non-melanogenic melanoma cells. The main target of this toxicity appears to be the mitochondria, which become markedly swollen and vacuolated. The mechanism of their action has been thought to be due to interference with oxidoreductases in the mitochondria. However, our results suggest that this cytotoxicity most probably does not result simply from inhibition of mitochondrial enzymes, but is closely related to the melanin biosynthesis pathway.

Normal uveal melanocytes of rhesus and cynomolgus macaques can be grown in culture for 3-9 months and subcultured a few times. Postnatal and adult choroidal melanocytes are terminally differentiated cells. They are melanin-containing but not actively melanin-synthesizing cells. They do not undergo cell division, nor do they incorporate tritiated thymidine, but otherwise they are metabolically active. Postnatal and young adult iridial melanocytes are metabolically more active than choroidal cells. They require a feeder cell layer for attachment and to be maintained in a healthy condition. An endothelial cell line established from a rhesus fetal choroid-retina proves to be an effective feeder layer for adult iridial cells. Fetal uveal melanocytes divide slowly and usually require some stimulus and a special culture environment supplemented with 12-O-tetradecanolphorbol-13-acetate and cholera toxin. They can grow and differentiate in vitro. Iridial melanocytes grow and change into cells resembling postnatal choroidal melanocytes. Similar changes occur during development in utero. These findings further suggest that, in vivo, iridial melanocytes migrate and mature to become choroidal melanocytes.

No abstract

The effects of theophylline and melanocyte-stimulating hormone (MSH) on cultured uveal melanocytes from the eyes of normal adult rhesus macaques were studied by light and electron microscopy and by dopa cytochemistry. The principal effects were changes in melanosome ultrastructure and an increased complexity of Golgi-associated vesicles and cisternae filled with dopa reaction products. The changes were more extensive in iris cells and less remarkable in choroid cells. The effects of theophylline were more pronounced than those of MSH. Our data suggest that normal iridial melanocytes, as do melanogenic murine melanoma cells, respond to theophylline or MSH by increasing tyrosinase synthesis, tyrosinase transfer, and melanization.