Effects of T3 and rearing temperature on growth and skeletal myosin heavy chain isoform transition during early development in the salmonid Salvelinus alpinus (L.)

Rearing of 1-year-old Arctic charr (Salvelinus alpinus) at 12°C, as well as the administration of 50 or 75 mgT₃/kg feed, accelerated the neonatal to adult fast myosin heavy chain transition, but the effect of temperature was more dramatic than the effect of T₃ administration. The endogenous plasma levels of T₃ in charrs reared at 12°C were higher than those of analogous groups reared at natural temperature, which in the period under study was between 0.5 and 12°C. As in other species, T₃ seemed to play a role in the regulation of the neonatal to adult fast myosin isoform transition by down-regulating the levels of the neonatal and increasing the levels of an adult fast myosin heavy chain. Temperature seemed to accelerate this transition at least, but not only, by inducing an increase in the endogenous levels of T₃ in the Arctic charr.     

Distinct structural elements of rab5 define its functional specificity.

Members of the rab family of small GTPases are localized to distinct cellular compartments and function as specific regulators of vesicle transport between organelles. Overexpression of rab5, which is associated with early endosomes and the plasma membrane, increases the rate of endocytosis [Bucci et al. (1992) Cell, 70, 715-728]. From sequence alignments and molecular modelling we identified structural elements that might contribute to the definition of the functional specificity of rab5. To test the role of these elements experimentally, we transplanted them onto rab6, which is associated with the Golgi complex. The chimeric proteins were assayed for intracellular localization and stimulation of endocytosis. First, we found that the C-terminus of rab5 could target rab6 to the plasma membrane and early endosomes but it did not confer rab5-like stimulation of endocytosis. Further replacement of other regions revealed that the N-terminus, helix alpha 2/loop 5 and helix alpha 2/loop 7 were all required to functionally convert rab6 into rab5. Reciprocal hybrids of rab5 containing these regions replaced with those of rab6 were inactive, demonstrating that each region is essential for rab5 function. These results indicate that distinct structural elements specify the localization, membrane association and regulatory function of rab5.     

SR141716A, a potent and selective antagonist of the brain cannabinoid receptor

SR141716A is the first selective and orally active antagonist of the brain cannabinoid receptor. This compound displays nanomolar affinity for the central cannabinoid receptor but is not active on the peripheral cannabinoid receptor. In vitro, SR141716A antagonises the inhibitory effects of cannabinoid receptor agonists on both mouse vas deferens contractions and adenylyl cyclase activity in rat brain membranes. After intraperitoneal or oral administration SR141716A antagonises classical pharmacological and behavioural effects of cannabinoid receptor agonists. This compound should prove to be a powerful tool for investigating the in vivo functions of the anandamide/cannabinoid system.     

Responses of salivary acinar cells to intracellular alkalinization

Responses of rat submandibular acini to intracellular alkalinization were investigated. Intracellular alkalinization was induced by addition of NH⁴Cl or methylamines, or by prepulse with Na butyrate. Only partial recovery occurred following Na butyrate prepulse or methylated amine addition, but full recovery was observed following addition of NH⁴Cl. The latter recovery was DIDS and dimethylamiloride-insensitive but was inhibited by bumetanide or high [K⁺] and stimulated in Na⁺ free buffer and by ouabain. Acetylcholine stimulated recovery from NH⁴Cl- or Na butyrate pre-pulse-induced alkalinization and reduced the extent of alkalinization induced by methylated amines. Acetylcholine-stimulated recovery from NH⁴Cl-induced alkalinization was mimicked by substance P or ionomycin and was partially Ca²⁺-dependent. This stimulated recovery was bumetanide-insensitive but was partially sensitive to charybdotoxin. Taken together, these data indicate that in unstimulated cells, recovery from alkalinization induced by NH⁴Cl occurs by bumetanide-sensitive transport of the NH⁴⁺ ion, that DIDS-inhibitable anion transport contributes little to this recovery, and that acetylcholine and other Ca²⁺-elevating agents accelerate recovery from NH⁴Cl-induced alkaline challenge by a mechanism insensitive to bumetanide, DIDS, ouabain, and dimethylamiloride but sensitive to extracellular Ca²⁺ and to charybdotoxin. Partial recovery from alkaline challenge can also occur in the absence of NH⁴⁺ ions, and acetylcholine also stimulates this mode of recovery. Together, these data suggest that these cells have little intrinsic ability to recover from intracellular alkalinization and that the NH⁴⁺ ion may be a surrogate for K⁺ in at least two ion transport pathways. © 1994 wiley-Liss, Inc.     

The role of ATP as a mediator in the action of iron complexes on cellular calcium homeostasis

The effects of the interaction between low molecular weight iron complexes (citrate, lactate, and ATP complexes) with ATP and proteins, on the modification of Ehrlich carcinoma cell calcium homeostasis have been studied. In that modification the ferric-ATP complex shows much higher activity than the others. Sodium ATP, by iron translocation from citrate and lactate, increases their activity. This phenomenon implicates ATP as a mediator on the cellular activity of the complexes. Proteins, particularly ferritin, appear to moderately reduce their activity, whereas glutathione and ascorbic acid, acting as lipid peroxidation-inhibitors, show only a slight reduction of the iron complex’s effects on cellular calcium uptake.     

Effect of various bud induction treatments on elongation and rooting of adventitious shoots of Canary Island pine (Pinus canariensis)

Three-day-old cotyledonary explants of Pinus canariensis were subjected to 30 induction treatments using half-strength Bornman's medium containing various combinations of N⁶- benzyladenine, zeatin, kinetin and 2-isopentenyl-adenine. The highest numbers of buds were obtained with 10 M 6-benzyladenine, but both kinetin and zeatin influenced shoot elongation. Shoots were maintained on half-strength Schenk and Hildebrandt medium with 2% sucrose and 0.05% activated charcoal. For rooting, shoots were pulsed for 4 h in a 100 M indole-3-butyric acid aqueous solution (pH 4.2–4.5), and planted in peat:vermiculite:perlite (1:1:1). After 8 weeks, the numbers of rooted shoots were similar for most treatments. Therefore, the bud induction treatments did not significantly influence rooting of adventitious shoots of Canary Island pine.     

Hepatitis B virus nucleocapsid particles do not cross the hepatocyte nuclear membrane in transgenic mice.

Transgenic mice that express the hepatitis B virus core protein were used to examine factors that influence the intracellular localization of nucleocapsid particles in the primary hepatocyte in vivo. In this model, viral nucleocapsid particles are strictly localized to the nucleus of the hepatocyte except when the nuclear membrane dissolves during cell division, at which time they enter the cytoplasm. The cytoplasmic nucleocapsid particles do not reenter the nucleus, however, when the nuclear membrane re-forms after cell division. The data support the notion that nucleocapsid particles can form de novo within the nucleus, and they suggest that performed nucleocapsid particles cannot be transported across the intact nuclear membrane in either direction. The results imply that nucleocapsid disassembly is probably required for entry of the hepadnaviral genome into the nucleus, and they question the role of the intranuclear viral nucleocapsid particle during the viral life cycle.     

Immobilization ofColeus blumei cells in a column reactor using a spray feeding system

Summary Coleus blumei cells were immobilized in a column reactor packed withLuffa cylindrica pieces. Medium was fed from the top of the column using a spray system and cells maintained high viability for 52 days. Cell growth was slower but rosmarinic acid production was better compared to immobilized cells in the shake flasks.     

Specific DNA probes for the identification of the fish pathogen,Renibacterium salmoninarum

To obtain specific DNA probes for the identification of the fish pathogen, Renibacterium salmoninarum, a discriminatory recombinant DNA library was constructed using selective fragments of the bacterial genome. Three renibacterial clones, pMAM29, pMAM46 and pMAM77, containing 149, 73, and 154 bp respectively, were isolated and characterized. The specificity of the probes was confirmed by dot-blot and Southern hybridization analyses. Bacterial hybridization experiments revealed that pMAM29 discriminates the R. salmoninarum genome from that of other fish pathogens such as Aeromonas salmonicida, Yersinia ruckeri, Flexibacter columnaris, Lactobacillus piscicola, Vibrio ordalii, Vibrio anguillarum and Aeromonas hydrophila. Thus, this probe may provide a new means to diagnose bacterial kidney disease in asymptomatic fish and ova.The authors are with the instituto de Bioquímica, Universidad Austral de Chile, P.O. Box 567, Valdivia, Chile     

Fish species composition before and after construction of a main stem reservoir on the White River,Colorado

Synopsis The completion in the fall of 1984 of Taylor Draw Dam on the White River, Colorado, formed Kenney Reservoir — thus impounding the last significant free-flowing tributary in the Upper Colorado River Basin. Fishes were sampled above and below the dam axis prior to closure of the dam and in the reservoir and river downstream following impoundment. While immediate effects of the dam to the ichthyofauna included blockage of upstream migration to 80 km of documented range for endangered Colorado squawfish, the reservoir also proved to have profound delayed effects on the river's species composition. Pre-impoundment investigations in 1983–1984 showed strong domination by native species above, within, and below the reservoir basin. By 1989–1990, non-native species comprised roughly 90% of the fishes collected in the reservoir and 80% of the fishes collected in the river below the dam. Initially, fathead minnow, whose numbers quickly increased in the new reservoir, dominated all post-impoundment collections, but red shiner became the most abundant fish collected in the river below the dam by 1989–1990. While agency stocking programs for the reservoir sought to emphasize a sport fishery for salmonids, primarily rainbow trout, local enthusiasm for warmwater sport fishes resulted in illicit transfers of these species from nearby impoundments. Several species, formerly rare or unreported in the White River in Colorado, including white sucker, northern pike, green sunfish, bluegill, largemouth bass and black crappie, were present in the river following impoundment. Our investigation indicates smaller-scale, main-stem impoundments that do not radically alter hydrologic or thermal regimes can still have a profound influence on native ichthyofauna by facilitating establishment and proliferation of nonnative species.Cooperators are the U.S. Fish and Wildlife, the Colorado Division of Wildlife, and Colorado State University