Organic osmolytes increase expression of specific tight junction proteins in skin and alter barrier function in keratinocytes.

BACKGROUND: The epidermal barrier is important for water conservation, failure of which is evident in dry-skin conditions. Barrier function is fulfilled by the stratum corneum, tight junctions (TJs, which control extracellular water) and keratinocyte mechanisms, such as organic osmolyte transport, which regulate intracellular water homeostasis. Organic osmolyte transport by keratinocytes is largely unexplored and nothing is known regarding how cellular and extracellular mechanisms of water conservation may interact. OBJECTIVES: We aimed to characterize osmolyte transporters in skin and keratinocytes, and, using transporter inhibitors, to investigate whether osmolytes can modify TJs. Such modification would suggest a possible link between intracellular and extracellular mechanisms of water regulation in skin. METHODS: Immunostaining and quantitative polymerase chain reaction of organic osmolyte-treated organ-cultured skin were used to identify changes to organic osmolyte transporters, and TJ protein and gene expression. TJ functional assays were performed on organic osmolyte-treated primary human keratinocytes in culture. RESULTS: Immunostaining demonstrated the expression of transporters for betaine, taurine and myo-inositol in transporter-specific patterns. Treatment of human skin with either betaine or taurine increased the expression of claudin-1, claudin-4 and occludin. Osmolyte transporter inhibition abolished this response. Betaine and taurine increased TJ function in primary human keratinocytes in vitro. CONCLUSIONS: Treatment of skin with organic osmolytes modulates TJ structure and function, which could contribute to the epidermal barrier. This emphasizes a role for organic osmolytes beyond the maintenance of intracellular osmolarity. This could be harnessed to enhance topical therapies for diseases characterized by skin barrier dysfunction.

Regulation of oxidative degradation of L-lysine in rat liver mitochondria.

The generation of 14CO2 from [1-14C]lysine by hepatic mitochondria through the saccharopine pathway is controlled by intramitochondrial concentrations of lysine, 2-oxoglutarate and NADPH. Mitochondria, isolated from rats pre-treated with glucagon, exhibited higher activities of L-lysine: 2-oxoglutarate reductase, saccharopine dehydrogenase and 2-aminoadipate aminotransferase. The flux through this pathway is stimulated in liver mitochondria after glucagon treatment. Multiple regulation of lysine oxidation in liver mitochondria confirms a complex mechanism of 'mitochondrial activation' by glucagon.

The effects of ovine growth hormone on protein turnover in rainbow trout.

Ovine growth hormone (oGH) was administered to rainbow trout via an intraperitoneal cholesterol implant. After 21 days, plasma oGH levels were recorded as control group, less than 2 ng ml-1, i.e., not detectable, and oGH group, 19.2 +/- 2.8 ng ml-1. oGH-treated fish exhibited significantly increased whole-body growth rates, whole-body protein accretion rates, stimulated tissue protein synthesis, and tissue protein accretion rates. A dramatic decrease in white muscle protein concentration was also observed after oGH treatment. In some tissues (liver and stomach), elevated protein synthesis rates were the result of higher RNA/protein ratios. However, in other tissues (gill and ventricle), increased RNA activity accounted for the differences in rates of protein synthesis. The growth promoting effects of oGH on both whole-body and tissue protein turnover were generally accompanied with no change in the efficiency of deposition of newly synthesized protein. For the same ration size, the oGH group showed higher retentions of ingested nitrogen. It is concluded that oGH significantly enhances whole-body growth rates as a result of the stimulatory effect on protein synthesis rates with little effect on protein degradation.