Cytokines induce dye coupling between cultured rat microglia.
(A and C) Phase-contrast views of the
fluorescence fields shown in B and D,
respectively. (B) An example of lack of dye coupling
observed under control conditions. (D) A cell that is
dye coupled to three neighboring cells in a microglia culture treated
with 1 ng/ml INF-γ plus 1 ng/ml TNF-α for 9 h.
(Bar, 80 μm.) (Graph) The incidence of dye coupling
(Lucifer yellow) was evaluated in cultures of rat microglia under
control conditions and after cytokine treatment for 9 h. Treatment
with 1 ng/ml INF-γ, 1 μg/ml LPS, or 1 ng/ml
TNF-α did not increase coupling above control levels. INF-γ (1
ng/ml) plus LPS (1 μg/ml) caused a large increase in
coupling, which was largely prevented by cotreatment with an
anti-TNF-α antibody. Treatment with 1 ng/ml INF-γ plus 1
ng/ml TNF-α also caused a large increase in coupling; this
coupling was blocked by 35 μM 18α-glycyrrhetinic acid (AGA), and
the block was reversed after the blocker was washed out (W/O
AGA). Each histogram bar corresponds to the mean ± SD of seven
experiments, in each of which a minimum of 10 cells were scored.