Phorbol ester stimulates hexose uptake by brain microvessel endothelial cells

Lester R Drewes, Margaret A. Broderius, David Z. Gerhart

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

Glucose uptake into cultured endothelial cells (EC) derived from brain microvessels was determined in the absence and presence of 12-O-tetradecanoylphorbol-13-acetate (TPA), EGTA, the calcium ionophore A23187, and insulin. EC were obtained from dog and human (autopsy) brain microvessels and maintained in culture for up to four passages. Monolayers of EC were treated with TPA and other compounds immediately prior to harvesting for hexose uptake measurements using 3-O-[3H]methyl-D-glucose, 2-[3H]deoxy-D-glucose, or D-[3H]glucose. Typically, treatment with TPA (0.1-100 ng/ml) resulted in hexose uptake levels 2 to 3 times those of controls, although occasionally levels 5 to 10 times those of controls were observed. Similar stimulation was observed with all radiolabeled hexoses. Stimulation by TPA was greatest in primary or first passage cells and was greatly diminished in older cells. Neither chelation of extracellular calcium with EGTA nor the presence of both EGTA and A23187 in the culture medium prevented the stimulatory effect of TPA. Insulin (1200 ng/ml) failed to stimulate hexose uptake. Treatment with 100 ng/ml TPA did not alter the appearance of actin filaments in canine EC as visualized with rhodamine phalloidin. These results, in combination with other recent studies, suggest that blood-brain glucose transport may be regulated by phorbol ester-activated protein kinase C.

Original languageEnglish (US)
Pages (from-to)771-776
Number of pages6
JournalBrain Research Bulletin
Volume21
Issue number5
DOIs
StatePublished - Nov 1988

Keywords

  • Brain
  • Dog
  • Endothelial cell culture
  • Glucose transport
  • Human
  • Microvessel
  • Tetradecanoylphorbol acetate

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