Abstract:
:The present study aimed to determine the kinetics of L-3, 4-dihydroxyphenylalanine (L-DOPA) uptake in an immortalised cell line of rat capillary cerebral endothelial cells (clones RBE 4 and RBE 4B), to define the type of inhibition produced by L-5-hydroxytryptophan (L-5-HTP), 2-aminobicyclo(2,2,1)-heptane-2-carboxylic acid (BHC) and N-(methylamino)-isobutyric acid (MeAlB) and its sodium dependence. Non-linear analysis of the saturation curves for L-DOPA and L-5-HTP revealed in RBE 4 cells Km values (in microM) of 72 and 102 and in RBE 4B cells Km values (in microM) of 60 and 118, respectively. IC50 values for L-5-HTP (RBE 4, 1026 microM; RBE 4B, 831 microM) obtained in the presence of a nearly saturating (250 microM) concentration of L-DOPA were almost 5-fold those obtained when non-saturating (25 microM) concentrations of L-DOPA were used. IC50 values for BHC obtained in the presence of a nearly saturating (250 microM) concentration of L-DOPA were also 6- to 5-fold those obtained when non-saturating (25 microM) concentrations of L-DOPA were used. MeAlB (up to 2.5 mM) was found not to interfere with the uptake of L-DOPA. In RBE 4 cells, Vmax values for L-DOPA uptake were identical in the absence and the presence of 150 microM L-5-HTP or 150 microM BHC, but Km values (microM) were significantly greater (P<0.05) when L-DOPA uptake was studied in the presence of L-5-HTP or BHC. Similar findings were observed when RBE 4B cells were used. Uptake of (250 microM) L-DOPA in the absence of sodium in the incubation medium was similar to that observed in the presence of increasing concentrations of sodium (20 to 140 mM). It is concluded that RBE 4 and RBE 4B cells are endowed with the L-type amino acid transporter through which L-DOPA and L-5-HTP can be taken up, and suggested that this immortalised cell line of rat capillary cerebral endothelium might constitute an interesting in vitro model for the study of BBB mechanisms, namely those concerning solute and nutrient transfer across the brain capillary endothelium.
journal_name
Brain Resjournal_title
Brain researchauthors
Gomes P,Soares-da-Silva Pdoi
10.1016/s0006-8993(99)01387-6subject
Has Abstractpub_date
1999-05-22 00:00:00pages
143-50issue
1-2eissn
0006-8993issn
1872-6240pii
S0006-8993(99)01387-6journal_volume
829pub_type
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