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  • Na+K+2Cl--cotransporter  (1)
  • alanine transamination pathway  (1)
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Cytotechnology 29 (1999), S. 45-53 
    ISSN: 1573-0778
    Keywords: alanine transamination pathway ; ammonium ions ; energy metabolism ; glutamine ; myeloma cells ; potassium ions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Potassium ions decrease the transport rate of ammonium ions into myeloma and hybridoma cells, one effect of the involved transport processes being an increased energy demand (Martinelle and Häggström, 1993; Martinelle et al., 1998b). Therefore, the effects of K+ and NH4+ on the energy metabolism of the murine myeloma cell line, Sp2/0-Ag14, were investigated. Addition of NH4Cl (10 mM) increased the metabolism via the alanine transaminase (alaTA) pathway, without increasing the consumption of glutamine. As judged by the alanine production, the energy formation from glutamine increased by 155%. The presence of elevated concentrations of KCl (10 mM) was positive, resulting in a decreased uptake of glutamine (45%), and an even larger suppression of ammonium ion formation (70%), while the same throughput via the alaTA pathway (and energy production from glutamine) was retained as in the control culture. However, the simultaneous presence of 10 mM K+ and 10 mM NH4+ was more inhibitory than NH4Cl alone; an effect that could not be ascribed to increased osmolarity. Although the culture with both K+ and NH4+ consumed 60% more glutamine than the culture with NH4+ alone, the energy generation from glutamine could not be increased further, due to the suppression of the glutamate dehydrogenase pathway. Furthermore, the data highlighted the importance of evaluating the metabolism via different energy yielding pathways, rather than solely considering the glutamine consumption for estimating energy formation from glutamine.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Cytotechnology 22 (1996), S. 251-254 
    ISSN: 1573-0778
    Keywords: ammonium ion trasport ; cell death ; Na+K+2Cl--cotransporter ; Na+/H+ exchanger ; bumetanide ; amiloride
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Ammonium can be transported into the cell by ion pumps in the cytoplasmic membrane. Ammonia then diffuse out through the cell membrane. A futile cycle is created that results in cytoplasmic acidification and extracellular alkalinisation. Ammonium transport can be quantified by measuring the extracellular pH changes occurring in a cell suspension (in PBS) after addition of ammonium. By using this technique, in combination with specific inhibitors of various ion pumps, it was shown that ammonium ions are transported across the cytoplasmic membrane by the Na+K+2Cl--cotransporter in both hybridoma and myeloma cells. Further, the Na+/H+ exchanger, which regulates intracellular pH by pumping out protons, was shown to be active during ammonium exposure. The viability of hybridoma cells suspended in PBS and exposed to NH inf4 sup+ for only 90 min, was reduced by 11% (50% necrosis and 50% apoptosis). A control cell suspension did not loose viability during this time. Turning off the activity of the Na+/H+ exchanger (by amiloride) during ammonium exposure decreased viability further, while inhibiting transport itself (by bumetanide) restored viability to the same level as for the control experiment with bumetanide alone. These results show that one effect of ammonia/ammonium on cell physiology is specifically related to the inward transport of ammonium ions by membrane bound ion pumps.
    Type of Medium: Electronic Resource
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