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Fig. 6. Gap junctions provide follicle-enclosed oocytes with access to
mechanisms that correct acidosis. SNARF-dextran was microinjected into
denuded and follicle-enclosed oocytes from day-10 mice. (A) Recovery
from acidosis was examined simultaneously in denuded and follicle-enclosed
oocytes from day-10 mice using SNARF-dextran in bicarbonate-free medium. The
graph shows a representative replicate of this experiment, in which six
follicle-enclosed (black traces) and five denuded oocytes were examined. The
follicle-enclosed oocytes recover from acidosis when Na+ is
replaced. The average rate of recovery following Na+-replacement is
significantly greater in the follicle-enclosed oocytes over the course of
three replicates (P<0.05). Note that the SNARF-dextran remains
restricted to the oocyte within the follicle (inset). (B) Recovery from
acidosis was examined in follicle-enclosed oocytes in the presence of DMSO
(left panel) or 150 µM AGA (centre), which were added during the experiment
at t=20 minutes. A representative replicate of each treatment group
is shown. (C) Analyses of the rate of pH recovery during the
Na+-free period and following Na+ replacement are shown.
Each bar represents the mean of three separate replicates, a total of 19-29
follicle-enclosed oocytes. Different letters above bars indicate significant
differences (ANOVA, P<0.01). Note that gap-junction inhibition
abrogates both Na+-dependent and Na+-independent phases
of acidosis recovery in follicle-enclosed oocytes. Note also that AGA does not
inhibit acidosis recovery in fully grown oocytes, or shells of granulosa cells
(B, right, one of two similar experiments is shown for each).
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