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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Journal of comparative physiology 165 (1996), S. 542-551 
    ISSN: 1432-136X
    Keywords: Starvation ; Respiratory quotient ; Nitrogen quotient ; Fuel ; Rainbow trout, Oncorhynchus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Oxygen consumption, CO2 excretion, and nitrogenous waste excretion (75% ammonia-N and 25% urea-N) were measured daily in 4-g rainbow trout over a 15-day starvation period. Oxygen consumption and CO2 excretion declined while N excretion increased transiently in the mid-part of the starvation period but was unchanged from control levels at the end. Component losses (as percentage of total fuel used) of protein, lipid, and carbohydrate were 66.5, 31.1, and 2.4% respectively, as measured from changes in body weight and body composition, the latter relative to a control group at day 0. Instantaneous fuel use, as calculated from the respiratory quotients and nitrogen quotients, indicated that relative protein use rose during starvation, but contributed at most 24% of the aerobic fuel (as carbon). Lipid metabolism fell from about 68 to 37%, and was largely replaced by carbohydrate metabolism which rose from 20 to 37%. We conclude that the two approaches measure different processes, and that the instantaneous method is preferred for physiological studies. The compositional method is influence by greater error, and measures the fuels depleted, not necessarily burned, because of possible interconversion and excretion of fuels.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Journal of comparative physiology 166 (1996), S. 37-45 
    ISSN: 1432-136X
    Keywords: Fish gills ; Epithelial cells ; Intracellular pH ; Na−H exchange ; Trout, Oncorhynchus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Using primary cultures of gill pavement cells from freshwater rainbow trout, a method is described for achieving confluent monolayers of the cells on glass coverslips. A continuous record of intracellular pH was obtained by loading the cells with the pH-sensitive flourescent dye 2′,7′-bis(2-carboxyethyl)-5(6)-carboxyfluorescein and mounting the coverslips in the flowthrough cuvette of a spectrofluorimeter. Experiments were performed in HEPES-buffered media nominally free of HCO3. Resting intracellular pH (7.43 at extracellular pH=7.70) was insensitive to the removal of Cl− or the application of 4-acetamido-4′-isothiocyanatostilbene-2,2′-disulfonic acid (0.1 mmol·l−1), but fell by about 0.3 units when Na+ was removed or in the presence of amiloride (0.2 mmol·l−1). Exposure to elevated ammonia (“ammonia prepulse”; 30 mmol·l−1 as NH4Cl for 6–9 min) produced an increase in intracellular pH (to about 8.1) followed by a slow decay, and washout of the pulse caused intracellular pH to fall to about 6.5. Intracellular non-HCO 3 − buffer capacity was about 13.4 slykes. Rapid recovery of intracellular pH from intracellular acidosis induced by ammonia prepulse was inhibited more than 80% in Na+-free conditions or in the presence of amiloride (0.2 mmol·l−1). Neither bafilomycin A1 (3 μmol·l−1) nor Cl removal altered the intracellular pH recovery rate. The K m for Na+ of the intracellular pH recovery mechanism was 8.3 mmol·l−1, and the rate constant at V max was 0.008·s−1 (equivalent to 5.60 mmol H+·l−1 cell water·min−1), which was achieved at external Na+ levels from 25 to 140 mmol·l−1. We conclude that intracellular pH in cultured gill pavement cells in HEPES-buffered, HCO 3 − -free media, both at rest and during acidosis, is regulated by a Na+/H+ antiport and not by anion-dependent mechanisms or a vacuolar H+-ATPase.
    Type of Medium: Electronic Resource
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