Thiamine deficiency in cultured neuroblastoma cells: effect on mitochondrial function and peripheral benzodiazepine receptors.
When neuroblastoma cells were transferred to a medium of low (6 nM) thiamine concentration, a 16-fold decrease in total intracellular thiamine content occurred within 8 days. Respiration and ATP levels were only slightly affected, but addition of a thiamine transport inhibitor (amprolium) decreased ATP content and increased lactate production. Oxygen consumption became low and insensitive to oligomycin and uncouplers. At least 25% of mitochondria were swollen and electron translucent. Cell mortality increased to 75% within 5 days. [3H]PK 11195, a specific ligand of peripheral benzodiazepine receptors (located in the outer mitochondrial membrane) binds to the cells with high affinity (KD = 1.4 +/- 0.2 nM). Thiamine deficiency leads to an increase in both Bmax and KD. Changes in binding parameters for peripheral benzodiazepine receptors may be related to structural or permeability changes in mitochondrial outer membranes. In addition to the high-affinity (nanomolar range) binding site for peripheral benzodiazepine ligands, there is a low-affinity (micromolar range) saturable binding for PK 11195. At micromolar concentrations, peripheral benzodiazepines inhibit thiamine uptake by the cells. Altogether, our results suggest that impairment of oxidative metabolism, followed by mitochondrial swelling and disorganization of cristae, is the main cause of cell mortality in severely thiamine-deficient neuroblastoma cells.
Bettendorff
L
L
Laboratory of Neurochemistry, University of Liége, Belgium.
Goessens
G
G
Sluse
F
F
Wins
P
P
Bureau
M
M
Laschet
J
J
Grisar
T
T
D000255:Adenosine Triphosphate / Q000378:metabolism; D000670:Amprolium / Q000494:pharmacology; D000818:Animals; D016923:Cell Death; D003470:Culture Media; D007546:Isoquinolines / Q000378:metabolism; D007773:Lactates / Q000096:biosynthesis; D019344:Lactic Acid; D051379:Mice; D008854:Microscopy, Electron; D008928:Mitochondria / Q000502:physiology*; D008933:Mitochondrial Swelling; D009447:Neuroblastoma / Q000378:metabolism* / Q000648:ultrastructure; D010101:Oxygen Consumption; D011963:Receptors, GABA-A / Q000378:metabolism*; D013831:Thiamine / Q000008:administration & dosage* / Q000378:metabolism; D014407:Tumor Cells, Cultured
D003470:Culture Media; D007546:Isoquinolines; D007773:Lactates; D011963:Receptors, GABA-A; D019344:Lactic Acid; D000255:Adenosine Triphosphate; D000670:Amprolium; D013831:Thiamine; C037850:PK 11195