The use of a prostacyclin analogue, [3H]iloprost,for studying prostacyclin-binding sites on human platelets and neuronal hybrid cells

The stable prostacyclin analogue [³H]iloprost has been used for labelling prostacyclin-binding sites on human platelets and NCB-20 neuronal hybrid cells. The ligand-binding properties of the sites have been determined and correlate well with stimulation of cAMP synthesis in NCB-20 cells and inhibition of aggregation in human platelets.     

Effect of cell swelling on membrane and cytoplasmic distribution of pICln

pI_Cln is found ubiquitouslyin mammalian cells and is postulated to play a critical role in cellvolume regulation. Mutagenesis studies led to the proposal thatpI_Cln is a swelling-activated anion channel. However, recent studies in Madin-Darby canine kidney cells and endothelial cells have shown that the protein is localized primarily to the cytoplasm. It has therefore been postulated that activation involves reversible translocation ofpI_Cln from the cytoplasm andinsertion into the plasma membrane. We tested this hypothesis usingseveral different approaches. Fractionation of C₆ glioma cells into plasmamembrane- and cytoplasm-containing fractions demonstrated that ~90%of the recovered pI_Cln wasconfined to the cytosol. Swelling had no effect on the relative amount of protein present in the plasma membrane fraction. Immunofluorescence microscopy revealed that pI_Cln islocalized primarily, if not exclusively, to the cytoplasm of swollenand nonswollen cells. Similarly, transfection of cells with a greenfluorescent protein-labeled pI_Clnconstruct failed to reveal any membrane localization of the protein.These findings do not support the hypothesis that pI_Cln is a volume regulatory anionchannel activated by swelling-induced membrane insertion.

     

Factors influencing the behavior and duration of residence of adult Chinook salmon in a stratified estuary

The duration of residence, behavior, and thermal experience of adult Chinook salmon (Oncorhynchus tshawytscha) in a stratified lagoon-type estuary over a four-year period was determined using acoustic and radio biotelemetry transmitters and archival temperature tags. Results did not support the hypothesis that adult Chinook salmon would hold extensively and migrate slowly through the estuarine lagoon with return trips to the sea and use of the salt wedge for behavioral thermo-osmoregulation in response to high summer water temperatures. Passive tidal transport was not observed as predicted and entry from the sea to the estuarine lagoon occurred during all tidal phases suggesting that conserving energy was not a priority for fish during the estuarine phase of their migration. An unexpected finding was that the persistent pursuit pressure of pinnipeds, especially from California sea lions (Zalophus californianus), appeared to negate the suitability of estuarine lagoon as holding habitat. This finding led to the hypothesis that the risk of pinniped predation was primarily responsible for the brief residence (<24 h on average) and rapid migration (mean 1.2 km/h, 0.42 body lengths/s) of adult Chinook salmon in the estuarine lagoon, especially given its relatively small size (7 km). This hypothesis will be difficult to test without exclusion of pinnipeds. Thermal records of fish that retreated back to the sea after tagging suggested that use of the marine river plume could be important for osmotic adaption to allow rapid migration through estuaries, which often contain concentrations of predators anticipating the return of migrating salmon. Alternatively, use of the marine river plume could have occurred independent of predators. Fish retreating back to the sea should be considered a tagging effect as downstream movement commonly occurs when adult salmonids are released after tagging. There was no evidence of any other tagging effects or biased behavior by fish tagged in this study. Regardless of the reasons, the successful migration of fish that held for weeks in the sea after tagging indicates that there could be considerable flexibility in the river entry timing of some salmon stocks. Tagging adult salmonids in the sea as they approach their natal rivers is ideal but the associated logistical challenges and expense make tagging fish immediately upon entrance to river mouths the next best option when possible.     

Studies on the relationship between glutathione S-transferase phenotype and bile acid binding by human liver cytosol

The possibility that the GST1 phenotype of human liver cytosol is a determinant of bile salt binding has been investigated by using equilibrium dialysis and gel-exclusion chromatography. Binding of bile salts was non-saturable and whereas the glutathione S-transferases did not appear to be major bile salt binders, other binding components with molecular weights of 35 000 and 11 000 were identified in both fetal and adult cytosols.