title: Choline uptake by cerebral capillary endothelial cells in culture.
abstract: A passage of choline from blood to brain and vice versa has been demonstrated in vivo. Because of the presence of the blood-brain barrier, such passage takes place necessarily through endothelial cells. To get a better understanding of this phenomenon, the choline transport properties of cerebral capillary endothelial cells have been studied in vitro. Bovine endothelial cells in culture were able to incorporate [3H]choline by a carrier-mediated mechanism. Nonlinear regression analysis of the uptake curves suggested the presence of two transport components in cells preincubated in the absence of choline. One component showed a Km of 7.59 +/- 0.8 microM and a maximum capacity of 142.7 +/- 9.4 pmol/2 min/mg of protein, and the other one was not saturable within the concentration range used (1-100 microM). When cells were preincubated in the presence of choline, a single saturable component was observed with a Km of 18.5 +/- 0.6 microM and a maximum capacity of 452.4 +/- 42 pmol/2 min/mg of protein. [3H]Choline uptake by endothelial cells was temperature dependent and was inhibited by the choline analogs hemicholinium-3, deanol, and AF64A. The presence of ouabain or 2,4-dinitrophenol did not affect the [3H]choline transport capacity of endothelial cells. Replacement of sodium by lithium and cell depolarization by potassium partially inhibited choline uptake. When cells had been preincubated without choline, recently transported [3H]choline was readily phosphorylated and incorporated into cytidine-5'-diphosphocholine and phospholipids; however, under steady-state conditions most (63%) accumulated [3H]choline was not metabolized within 1 h.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Pathology, UCLA School of Medicine.
authors.forename: C
authors.initials: C
authors.lastname: Estrada
authors.forename: J
authors.initials: J
authors.lastname: Bready
authors.forename: J
authors.initials: J
authors.lastname: Berliner
authors.forename: P A
authors.initials: PA
authors.lastname: Cancilla
mesh_terms: D000818:Animals; D001692:Biological Transport; D002196:Capillaries; D002478:Cells, Cultured; D002560:Cerebrovascular Circulation; D002794:Choline / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D009994:Osmolar Concentration; D012044:Regression Analysis
chemical_list: D002794:Choline