The pathways of lead (Pb2+) uptake were studied in fura-2-loaded cerebellar
granule cells from 8-day-old rats. In a nominal Ca-free external bath, Pb2
+ (5-50 muM) determined an increase of the fluorescence emission ratio (R =
E-340/E-380) even in the absence of any specific stimulus. This rise was d
ose-dependent, was not significantly affected by mM Mg2+ or Ca2+, but it wa
s readily reversed by the membrane-permeant heavy metal chelator tetrakis(2
-pyridylmethyl) ethylenediamine (TPEN, 100 muM), indicating that it was due
to Pb2+ influx. The rate of rise, dR/dt, was increased up to a factor of 5
by depolarizing high-KCl solution, indicating a sizeable permeation throug
h voltage-dependent channels. This effect was neither antagonized by nimodi
pine, nor enhanced by BayK8644, but it was slackened by omega -agatoxin IVA
(200 nm), suggesting an involvement of non-L-type calcium channels. Pb2+ i
nflux was also stimulated by glutamic acid or NMDA in the presence of 10-30
muM glycine, but only in Mg-free solution, suggesting that glutamate chann
els of the NMDA type are an additional pathway of Pb2+ uptake. Pb2+ caused
a time-, dose- and stimulus-dependent saturation of the dye, whose intracel
lular concentration is similar to 10 muM, indicating that intracellular Pb2
+ can readily reach a concentration in the micromolar range. These results
indicate that the particular vulnerability of neurones to Pb2+ poisoning is
linked to the presence of specific transport systems, which mediate the ra
pid uptake of Pb2+ into the neurone.