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Fig. 2. Glucose, pyruvate and lactate have distinct effects on NAD(P)H and
FAD++ autofluorescence, indicating distinct metabolic pathways.
(A) Timecourse of changes in NAD(P)H (blue) and FAD++
(green) autofluorescence in a MII oocyte incubated at time 0 in H-KSOM/AA that
was devoid of glucose, lactate and pyruvate (n=35 oocytes). These
substrates were added at the times indicated at the bottom of the graph.
(B) Inhibition of the pentose phosphate pathway: timecourse of changes
in autofluorescence in a fully grown GV oocyte incubated in complete H-KSOM/AA
(n=12 oocytes). The PPP inhibitor DHEA was added to the chamber at
the time indicated at the top of the graph. Continuous traces indicate global
signals (large, dark-blue ROI in the FAD++ image). Dashed traces
indicate nuclear signals (small, light-blue ROI in the FAD++ image,
left). (C) Inhibition of LDH: timecourse of changes in autofluorescence
in a mature MII oocyte incubated in H-KSOM/AA without lactate or pyruvate
(n=18 oocytes). Lactate (2 mmol/l) was added, as indicated, followed
by the addition of the LDH inhibitor, oxamate (10 mmol/l). The concentration
of lactate in the medium was then increased to 8 mmol/l, as indicated. The
x-axes indicate time in minutes.
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