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J Neurophysiol 76: 764-769, 1996;
0022-3077/96 $5.00
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Journal of Neurophysiology, Vol 76, Issue 2 764-769, Copyright © 1996 by APS

ARTICLES

Swelling-activated amino acid efflux in the human neuroblastoma cell line CHP-100

S. Basavappa, C. C. Huang, A. W. Mangel, D. V. Lebedev, P. A. Knauf and J. C. Ellory
University Laboratory of Physiology, University of Oxford, United Kingdom.

1. The effects of hypoosmotic stress on cell volume and amino acid efflux were evaluated in the human neuroblastoma cell line CHP-100 with the Coulter Counter Multisizer and radiolabeled amino acid efflux, respectively. 2. CHP-100 cells swelled by approximately 35 +/- 5% (means +/- SE) when the osmolarity of the solution was decreased from 290 to 190 mOsm/kg H2O. The rapid swelling was followed by a biphasic regulatory volume decrease (RVD). 3. In cells loaded with 14C-taurine, hypoosmotic stress induced a 300 +/- 22% (n = 23, P < 0.05) increase in taurine efflux compared with controls. This efflux was inhibited by the chloride channel blockers 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB), 4,4'-diisothio-cyanostilbene-2,2'-disulfonic acid (DIDS), niflumic acid and by the volume-activated anion channel blocker tamoxifen. In addition, the swelling-activated taurine efflux was dependent upon extracellular calcium. 4. Similarly, in cells loaded with 14C-glycine, hypoosmotic stress significantly increased glycine efflux, which was also sensitive to NPPB. In contrast, efflux of 3H-glutamate was not significantly altered after hypoosmotic stress. 5. With the use of patch clamp recording techniques, Cl- channels were activated in cell attached patches after exposure to hypoosmotic solutions. 6. In nystatin perforated patches, permeability of the hypoosmotically activated anion channel was observed to be SCN- > I- > Br- > Cl- >> Glutamate. 7. It is concluded that in CHP-100 cells, anion channels are activated during hypoosmotic stress and these channels represent a pathway for efflux of amino acids.


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S. Basavappa, S. F. Pedersen, N. K. Jorgensen, J. C. Ellory, and E. K. Hoffmann
Swelling-Induced Arachidonic Acid Release via the 85-kDa cPLA2 in Human Neuroblastoma Cells
J Neurophysiol, March 1, 1998; 79(3): 1441 - 1449.
[Abstract] [Full Text] [PDF]


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