Treatment of human melanocyte cell cultures with the alpha-2 adrenergic receptor antagonist yohimbine results in a marked down-regulation of tyrosinase activity. A 30% decrease occurs within 12 h of exposure of cells to yohimbine (100 microM), and by 48 h tyrosinase activity in treated melanocytes is less than a fifth that of control cultures. The inhibition is dose dependent and occurs in human melanocytes derived from either black or white skin types, and also in mouse melanoma cells. The yohimbine-induced decrease in tyrosinase activity is reversible, with enzyme levels returning to 90% of control values 48 h after removal of drug. Although tyrosinase activity is markedly suppressed by yohimbine, the compound has no effect on cell proliferation, cellular translation, or DNA synthesis. Treatment of melanocyte cultures with yohimbine blocks the increase in tyrosinase activity by either 3-isobutyl-1-methylxanthine, dibutyryl cAMP, or forskolin. Results of cAMP immunoassays, show that intracellular levels of the cyclic nucleotide are unaffected in cells treated with yohimbine. Tyrosinase inhibition by yohimbine does not involve a decrease in substrate availability since tyrosine uptake studies show that yohimbine has no effect on the amount of tyrosine entering the cell. Incubation of a melanosome-enriched fraction with yohimbine does not cause a lowering of tyrosinase activity, suggesting that an intact cell is required for yohimbine action. In addition, tyrosinase extracts show no reduction in activity when incubated directly with yohimbine, indicating that the drug does not act as a direct inhibitor of the enzyme. Finally, results of western immunoblotting show that yohimbine does not significantly lower the amount of tyrosinase protein in human melanocytes. These findings suggest that yohimbine acts through an as yet unidentified signaling pathway to lower the catalytic activity of pre-existing tyrosinase molecules present in melanocytes.

Glial fibrillary acid protein (GFAP) and vimentin are type III intermediate filament proteins, ubiquitously expressed in retinal glial cells. Under retinal stress, both GFAP and vimentin are well-known sensitive markers for retinal gliosis. However, little is known about whether these proteins are released into the vitreous body in response to retinal gliosis or are related to the severity of retinal gliosis seen in proliferative vitreoretinopathy (PVR). METHODS. Vitreous fluids were collected from 44 patients who underwent pars plana vitrectomy for macular hole (Group 1; n = 8), epiretinal membrane (Group 2; n = 8), or retinal detachment (RD) with various degrees of PVR (Group 3; n = 28). The severity of PVR was determined by cumulative scores using PVR classification. GFAP, vimentin, and total protein levels from the vitreous samples were measured. RESULTS. Both GFAP and vimentin levels were significantly elevated in vitreous fluid from Group 3 (RD) compared with Groups 1 and 2 (P < 0.01). GFAP levels (ng/mL) were 12.4