Validation of red cell sodium-lithium countertransport measurement--influence of different loading conditions

TY - JOUR

T1 - Validation of red cell sodium-lithium countertransport measurement--influence of different loading conditions

AU - Besch, W

AU - Schläger, D

AU - Brahm, J

AU - Kohnert, K D

PY - 1995/10

Y1 - 1995/10

N2 - Increased sodium-lithium countertransport in erythrocytes from patients with long-standing type I (insulin-dependent) diabetes mellitus has been considered as an early marker of nephropathy. Since the activity and kinetics of the sodium-lithium countertransport may critically depend on loading conditions, this study was aimed at determining sodium-lithium countertransport activity, Michaelis constant Km and maximum velocity Vmax in erythrocytes loaded in two different Li+ solutions. Sodium-lithium countertransport activity was determined in erythrocytes in 8 healthy control subjects after loading with 150 mmol/l LiCl compared with those loaded with 150 mmol/l LiHCO3. Sodium-lithium countertransport activity was similar for both loading procedures, although the erythrocyte lithium content did significantly differ (mean +/- SEM, 7.0 +/- 0.5 for LiCl and 8.9 +/- 0.5 mmol/l of cells for 150 mmol/l LiHCO3). There were no significant changes in the Km and Vmax. Increase of osmolality in efflux media containing 200 and 250 mmol/l NaCl resulted in a negligible shrinking of the red blood cells, not exceeding 2.2%. The main advantage is the short loading time of 15 min for LiHCO3 compared with 3 hours for LiCl. Under these conditions saturating intracellular Li+ concentrations can be obtained much more rapidly than with LiCl loading, thereby minimising alterations of the cell membranes. LiHCO3 loading shortens the experimental time considerably and enables a greater number of samples to be screened from larger population cohorts.

AB - Increased sodium-lithium countertransport in erythrocytes from patients with long-standing type I (insulin-dependent) diabetes mellitus has been considered as an early marker of nephropathy. Since the activity and kinetics of the sodium-lithium countertransport may critically depend on loading conditions, this study was aimed at determining sodium-lithium countertransport activity, Michaelis constant Km and maximum velocity Vmax in erythrocytes loaded in two different Li+ solutions. Sodium-lithium countertransport activity was determined in erythrocytes in 8 healthy control subjects after loading with 150 mmol/l LiCl compared with those loaded with 150 mmol/l LiHCO3. Sodium-lithium countertransport activity was similar for both loading procedures, although the erythrocyte lithium content did significantly differ (mean +/- SEM, 7.0 +/- 0.5 for LiCl and 8.9 +/- 0.5 mmol/l of cells for 150 mmol/l LiHCO3). There were no significant changes in the Km and Vmax. Increase of osmolality in efflux media containing 200 and 250 mmol/l NaCl resulted in a negligible shrinking of the red blood cells, not exceeding 2.2%. The main advantage is the short loading time of 15 min for LiHCO3 compared with 3 hours for LiCl. Under these conditions saturating intracellular Li+ concentrations can be obtained much more rapidly than with LiCl loading, thereby minimising alterations of the cell membranes. LiHCO3 loading shortens the experimental time considerably and enables a greater number of samples to be screened from larger population cohorts.

KW - Adult

KW - Antiporters

KW - Cells, Cultured

KW - Erythrocytes

KW - Humans

KW - Lithium

KW - Reproducibility of Results

KW - Sodium

KW - Time Factors

M3 - Journal article

C2 - 8608193

SN - 0939-4974

VL - 33

SP - 715

EP - 719

JO - European Journal of Clinical Chemistry and Clinical Biochemistry

JF - European Journal of Clinical Chemistry and Clinical Biochemistry

IS - 10

ER -