Sphingolipid long-chain-base auxotrophs of Saccharomyces cerevisiae: genetics, physiology, and a method for their selection.

A selection method for sphingolipid long-chain-base auxotrophs of Saccharomyces cerevisiae was devised after observing that strains that require a long-chain base for growth become denser when starved for this substance. Genetic analysis of over 60 such strains indicated only two complementation classes, lcb1 and lcb2. Mutant strains from each class grew equally well with 3-ketodihydrosphingosine, erythrodihydrosphingosine or threodihydrosphingosine, or phytosphingosine. Since these metabolites represent the first, second, and last components, respectively, of the long-chain-base biosynthetic pathway, it is likely that the LCB1 and LCB2 genes are involved in the first step of long-chain-base synthesis. The results of long-chain-base starvation in the Lcb- strains suggest that one or more sphingolipids have a vital role in S. cerevisiae. Immediate sequelae of long-chain-base starvation were loss of viability, exacerbated in the presence of alpha-cyclodextrin, and loss of phosphoinositol sphingolipid synthesis but not phosphatidylinositol synthesis. Loss of viability with long-chain-base starvation could be prevented by also blocking either protein or nucleic acid synthesis. Without a long-chain-base, cell division, dry mass accumulation, and protein synthesis continued at a diminished rate and were further inhibited by the detergent Tergitol. The cell density increase induced by long-chain-base starvation is thus explained as a differential loss of cell division and mass accumulation. Long-chain-base starvation in Lcb- S. cerevisiae and inositol starvation of Inos- S. cerevisiae share common features: an increase in cell density and a loss of cell viability overcome by blocking macromolecular synthesis.     

The use of viable hepatocytes to study the hormonal control of glycogenolysis in the chicken

Summary The rapid isolation of high yields of parenchymal cells from chicken liver is described. Stringent tests of viability show that the isolated hepatocytes are both structurally and metabolically similar to those in intact liver. During incubation viability decreased and the significance of this change on the interpretation of metabolic experiments is discussed. Lactate was a much more effective gluconeogenic precursor than pyruvate even in the presence of additional reducing equivalents. Hepatocytes isolated from fed chickens produced glucose from glycogen degradation. Glycogenolysis was stimulated by glucagon, dibutyryl cyclic AMP and adrenaline. Half maximal glucagon effects were elicited by physiological concentrations of the hormone. Glucagon and dibutyryl cyclic AMP stimulated glucagon, dibutyryl cyclic AMP and adrenaline their action was not additive to that of adrenaline.     

Tissue metal concentrations in two crayfish species cohabiting a tennessee cave stream

Summary Concentrations of Cd, Cr, Cu, Fe, Mg, Mn, Pb, Zn, Ca and K were examined in tissues of the troglobitic (obligatory cave-dwelling) crayfish Orconectes australis australis and troglophilic (facultative cave-dwelling) species Cambarus tenebrosus. These two species cohabit a stream in Merrybranch Cave, located in rural White Co., Tennessee. Tissue concentrations of essential metals did not exhibit any trends between species. In contrast, Cd and Pb concentrations were found to be significantly greater in O. a. australis for almost all of the tissues analyzed. The higher Cd and Pb concentrations in O. a. australis are thought to be due to the increased longevity of this troglobitic species. Because of the toxicity of Cd and Pb, chronic exposure to relatively low concentrations of these metals could cause changes in mortality, fecundity or behavior in aquatic organisms possessing long life spans. The bioaccumulation of metals from low level, non-point sources is discussed in relation to life history strategies.